KR20150132813A - 리파아제 활성이 증대된 양친매성 펩타이드-리파아제 결합체 및 이의 용도 - Google Patents
리파아제 활성이 증대된 양친매성 펩타이드-리파아제 결합체 및 이의 용도 Download PDFInfo
- Publication number
- KR20150132813A KR20150132813A KR1020150155169A KR20150155169A KR20150132813A KR 20150132813 A KR20150132813 A KR 20150132813A KR 1020150155169 A KR1020150155169 A KR 1020150155169A KR 20150155169 A KR20150155169 A KR 20150155169A KR 20150132813 A KR20150132813 A KR 20150132813A
- Authority
- KR
- South Korea
- Prior art keywords
- gly
- leu
- ala
- ser
- lys
- Prior art date
Links
- 239000004367 Lipase Substances 0.000 title claims abstract description 373
- 235000019626 lipase activity Nutrition 0.000 title abstract description 13
- 108090001060 Lipase Proteins 0.000 claims abstract description 299
- 102000004882 Lipase Human genes 0.000 claims abstract description 283
- 235000019421 lipase Nutrition 0.000 claims abstract description 280
- 239000003225 biodiesel Substances 0.000 claims abstract description 44
- 238000000034 method Methods 0.000 claims abstract description 44
- 239000002157 polynucleotide Substances 0.000 claims abstract description 27
- 150000002632 lipids Chemical class 0.000 claims abstract description 26
- 102000040430 polynucleotide Human genes 0.000 claims abstract description 26
- 108091033319 polynucleotide Proteins 0.000 claims abstract description 26
- 239000013604 expression vector Substances 0.000 claims abstract description 16
- 101800002011 Amphipathic peptide Proteins 0.000 claims abstract description 10
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 123
- 238000004519 manufacturing process Methods 0.000 claims description 43
- RZVAJINKPMORJF-UHFFFAOYSA-N Acetaminophen Chemical compound CC(=O)NC1=CC=C(O)C=C1 RZVAJINKPMORJF-UHFFFAOYSA-N 0.000 claims description 37
- 239000000758 substrate Substances 0.000 claims description 23
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 14
- 239000000203 mixture Substances 0.000 claims description 14
- 229920001519 homopolymer Polymers 0.000 claims description 6
- 239000002202 Polyethylene glycol Substances 0.000 claims description 5
- 229920001223 polyethylene glycol Polymers 0.000 claims description 5
- 230000008569 process Effects 0.000 claims description 5
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 4
- 241000105535 Photobacterium lipolyticum Species 0.000 claims description 4
- 239000002773 nucleotide Substances 0.000 claims description 4
- 125000003729 nucleotide group Chemical group 0.000 claims description 4
- 229920000642 polymer Polymers 0.000 claims description 4
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims description 3
- 241000589540 Pseudomonas fluorescens Species 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims description 3
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims description 2
- 229920002101 Chitin Polymers 0.000 claims description 2
- 229920002988 biodegradable polymer Polymers 0.000 claims description 2
- 239000004621 biodegradable polymer Substances 0.000 claims description 2
- 229920001577 copolymer Polymers 0.000 claims description 2
- 150000004676 glycans Chemical class 0.000 claims description 2
- 229920002674 hyaluronan Polymers 0.000 claims description 2
- 229960003160 hyaluronic acid Drugs 0.000 claims description 2
- 229920001583 poly(oxyethylated polyols) Polymers 0.000 claims description 2
- 229920001451 polypropylene glycol Polymers 0.000 claims description 2
- 229920001282 polysaccharide Polymers 0.000 claims description 2
- 239000005017 polysaccharide Substances 0.000 claims description 2
- 229920002451 polyvinyl alcohol Polymers 0.000 claims description 2
- 239000002904 solvent Substances 0.000 claims description 2
- 108010072641 thermostable lipase Proteins 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 9
- 241000907897 Tilia Species 0.000 claims 1
- 230000000593 degrading effect Effects 0.000 claims 1
- 235000019422 polyvinyl alcohol Nutrition 0.000 claims 1
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 108
- 108020004414 DNA Proteins 0.000 description 87
- 102000053602 DNA Human genes 0.000 description 87
- 108090000623 proteins and genes Proteins 0.000 description 69
- 108010050848 glycylleucine Proteins 0.000 description 67
- 108010041407 alanylaspartic acid Proteins 0.000 description 55
- QITBQGJOXQYMOA-ZETCQYMHSA-N Gly-Gly-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)CNC(=O)CN QITBQGJOXQYMOA-ZETCQYMHSA-N 0.000 description 47
- 108010047495 alanylglycine Proteins 0.000 description 43
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 40
- 239000012634 fragment Substances 0.000 description 38
- 239000013615 primer Substances 0.000 description 38
- 150000001413 amino acids Chemical class 0.000 description 37
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 34
- 108010078144 glutaminyl-glycine Proteins 0.000 description 31
- 230000027455 binding Effects 0.000 description 30
- 108010000434 glycyl-alanyl-leucine Proteins 0.000 description 30
- KRQSPVKUISQQFS-FJXKBIBVSA-N Arg-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCN=C(N)N KRQSPVKUISQQFS-FJXKBIBVSA-N 0.000 description 29
- NXSFUECZFORGOG-CIUDSAMLSA-N Ala-Asn-Leu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O NXSFUECZFORGOG-CIUDSAMLSA-N 0.000 description 26
- RLFSBAPJTYKSLG-WHFBIAKZSA-N Gly-Ala-Asp Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(O)=O RLFSBAPJTYKSLG-WHFBIAKZSA-N 0.000 description 26
- VSVZIEVNUYDAFR-YUMQZZPRSA-N Gly-Ala-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)CN VSVZIEVNUYDAFR-YUMQZZPRSA-N 0.000 description 26
- NWOSHVVPKDQKKT-RYUDHWBXSA-N Gly-Tyr-Gln Chemical compound [H]NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(O)=O NWOSHVVPKDQKKT-RYUDHWBXSA-N 0.000 description 26
- TVYWVSJGSHQWMT-AJNGGQMLSA-N Ile-Leu-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N TVYWVSJGSHQWMT-AJNGGQMLSA-N 0.000 description 26
- 108010079364 N-glycylalanine Proteins 0.000 description 26
- 108010054155 lysyllysine Proteins 0.000 description 26
- 108010005834 tyrosyl-alanyl-glycine Proteins 0.000 description 26
- XPJBQTCXPJNIFE-ZETCQYMHSA-N Gly-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)CN XPJBQTCXPJNIFE-ZETCQYMHSA-N 0.000 description 25
- 108010005233 alanylglutamic acid Proteins 0.000 description 25
- 230000000694 effects Effects 0.000 description 25
- 108010034529 leucyl-lysine Proteins 0.000 description 25
- 238000006243 chemical reaction Methods 0.000 description 24
- 238000003752 polymerase chain reaction Methods 0.000 description 24
- DSFYPIUSAMSERP-IHRRRGAJSA-N Leu-Leu-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N DSFYPIUSAMSERP-IHRRRGAJSA-N 0.000 description 23
- AIRZWUMAHCDDHR-KKUMJFAQSA-N Lys-Leu-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O AIRZWUMAHCDDHR-KKUMJFAQSA-N 0.000 description 22
- 230000001965 increasing effect Effects 0.000 description 22
- 102000004196 processed proteins & peptides Human genes 0.000 description 22
- 230000004927 fusion Effects 0.000 description 20
- DJIMLSXHXKWADV-CIUDSAMLSA-N Asn-Leu-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(N)=O DJIMLSXHXKWADV-CIUDSAMLSA-N 0.000 description 19
- LXKNSJLSGPNHSK-KKUMJFAQSA-N Leu-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N LXKNSJLSGPNHSK-KKUMJFAQSA-N 0.000 description 19
- 108010040443 aspartyl-aspartic acid Proteins 0.000 description 19
- 210000004027 cell Anatomy 0.000 description 19
- VKVDRTGWLVZJOM-DCAQKATOSA-N Leu-Val-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O VKVDRTGWLVZJOM-DCAQKATOSA-N 0.000 description 18
- YUAXTFMFMOIMAM-QWRGUYRKSA-N Lys-Lys-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O YUAXTFMFMOIMAM-QWRGUYRKSA-N 0.000 description 18
- SOAYQFDWEIWPPR-IHRRRGAJSA-N Met-Ser-Tyr Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O SOAYQFDWEIWPPR-IHRRRGAJSA-N 0.000 description 18
- 108010064235 lysylglycine Proteins 0.000 description 18
- 239000013598 vector Substances 0.000 description 18
- XNCUYZKGQOCOQH-YUMQZZPRSA-N Ser-Leu-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O XNCUYZKGQOCOQH-YUMQZZPRSA-N 0.000 description 17
- 235000001014 amino acid Nutrition 0.000 description 17
- 229940024606 amino acid Drugs 0.000 description 17
- 108010015792 glycyllysine Proteins 0.000 description 17
- 239000013612 plasmid Substances 0.000 description 17
- 108010080629 tryptophan-leucine Proteins 0.000 description 17
- XYTNPQNAZREREP-XQXXSGGOSA-N Ala-Glu-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XYTNPQNAZREREP-XQXXSGGOSA-N 0.000 description 16
- MNZHHDPWDWQJCQ-YUMQZZPRSA-N Ala-Leu-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O MNZHHDPWDWQJCQ-YUMQZZPRSA-N 0.000 description 16
- IHRGVZXPTIQNIP-NAKRPEOUSA-N Ala-Met-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@H](C)N IHRGVZXPTIQNIP-NAKRPEOUSA-N 0.000 description 16
- BHFOJPDOQPWJRN-XDTLVQLUSA-N Ala-Tyr-Gln Chemical compound C[C@H](N)C(=O)N[C@@H](Cc1ccc(O)cc1)C(=O)N[C@@H](CCC(N)=O)C(O)=O BHFOJPDOQPWJRN-XDTLVQLUSA-N 0.000 description 16
- AIJAPFVDBFYNKN-WHFBIAKZSA-N Gly-Asn-Asp Chemical compound C([C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)CN)C(=O)N AIJAPFVDBFYNKN-WHFBIAKZSA-N 0.000 description 16
- BHPQOIPBLYJNAW-NGZCFLSTSA-N Gly-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)CN BHPQOIPBLYJNAW-NGZCFLSTSA-N 0.000 description 16
- DRKZDEFADVYTLU-AVGNSLFASA-N His-Val-Val Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(O)=O DRKZDEFADVYTLU-AVGNSLFASA-N 0.000 description 16
- CYHJCEKUMCNDFG-LAEOZQHASA-N Ile-Gln-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)NCC(=O)O)N CYHJCEKUMCNDFG-LAEOZQHASA-N 0.000 description 16
- AVEGDIAXTDVBJS-XUXIUFHCSA-N Leu-Ile-Arg Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O AVEGDIAXTDVBJS-XUXIUFHCSA-N 0.000 description 16
- KPYAOIVPJKPIOU-KKUMJFAQSA-N Leu-Lys-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(O)=O KPYAOIVPJKPIOU-KKUMJFAQSA-N 0.000 description 16
- BMHIFARYXOJDLD-WPRPVWTQSA-N Met-Gly-Val Chemical compound [H]N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O BMHIFARYXOJDLD-WPRPVWTQSA-N 0.000 description 16
- CUMXHKAOHNWRFQ-BZSNNMDCSA-N Phe-Asp-Tyr Chemical compound C([C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=CC=C1 CUMXHKAOHNWRFQ-BZSNNMDCSA-N 0.000 description 16
- ZLXKLMHAMDENIO-DCAQKATOSA-N Pro-Lys-Asp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(O)=O ZLXKLMHAMDENIO-DCAQKATOSA-N 0.000 description 16
- IXCHOHLPHNGFTJ-YUMQZZPRSA-N Ser-Gly-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CO)N IXCHOHLPHNGFTJ-YUMQZZPRSA-N 0.000 description 16
- KZSYAEWQMJEGRZ-RHYQMDGZSA-N Thr-Leu-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O KZSYAEWQMJEGRZ-RHYQMDGZSA-N 0.000 description 16
- SCSVNSNWUTYSFO-WDCWCFNPSA-N Thr-Lys-Glu Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O SCSVNSNWUTYSFO-WDCWCFNPSA-N 0.000 description 16
- QNTBGBCOEYNAPV-CWRNSKLLSA-N Trp-Asn-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)N)C(=O)O QNTBGBCOEYNAPV-CWRNSKLLSA-N 0.000 description 16
- LGEYOIQBBIPHQN-UWJYBYFXSA-N Tyr-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 LGEYOIQBBIPHQN-UWJYBYFXSA-N 0.000 description 16
- 108010064997 VPY tripeptide Proteins 0.000 description 16
- JYPCXBJRLBHWME-UHFFFAOYSA-N glycyl-L-prolyl-L-arginine Natural products NCC(=O)N1CCCC1C(=O)NC(CCCN=C(N)N)C(O)=O JYPCXBJRLBHWME-UHFFFAOYSA-N 0.000 description 16
- XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 description 16
- 108010025801 glycyl-prolyl-arginine Proteins 0.000 description 16
- 108010079413 glycyl-prolyl-glutamic acid Proteins 0.000 description 16
- 108010044311 leucyl-glycyl-glycine Proteins 0.000 description 16
- 108010057821 leucylproline Proteins 0.000 description 16
- 108010025153 lysyl-alanyl-alanine Proteins 0.000 description 16
- 239000004006 olive oil Substances 0.000 description 16
- 235000008390 olive oil Nutrition 0.000 description 16
- 108010070409 phenylalanyl-glycyl-glycine Proteins 0.000 description 16
- 108010029020 prolylglycine Proteins 0.000 description 16
- 108010031491 threonyl-lysyl-glutamic acid Proteins 0.000 description 16
- 108010017949 tyrosyl-glycyl-glycine Proteins 0.000 description 16
- PCIFXPRIFWKWLK-YUMQZZPRSA-N Ala-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@H](C)N PCIFXPRIFWKWLK-YUMQZZPRSA-N 0.000 description 15
- UWIQWPWWZUHBAO-ZLIFDBKOSA-N Ala-Leu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](NC(=O)[C@H](C)N)CC(C)C)C(O)=O)=CNC2=C1 UWIQWPWWZUHBAO-ZLIFDBKOSA-N 0.000 description 15
- OXEMJGCAJFFREE-FXQIFTODSA-N Glu-Gln-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O OXEMJGCAJFFREE-FXQIFTODSA-N 0.000 description 15
- LGIMRDKGABDMBN-DCAQKATOSA-N Ser-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CO)N LGIMRDKGABDMBN-DCAQKATOSA-N 0.000 description 15
- JNKAYADBODLPMQ-HSHDSVGOSA-N Thr-Trp-Val Chemical compound C1=CC=C2C(C[C@@H](C(=O)N[C@@H](C(C)C)C(O)=O)NC(=O)[C@@H](N)[C@@H](C)O)=CNC2=C1 JNKAYADBODLPMQ-HSHDSVGOSA-N 0.000 description 15
- 108020001507 fusion proteins Proteins 0.000 description 15
- 102000037865 fusion proteins Human genes 0.000 description 15
- 239000003921 oil Substances 0.000 description 15
- 235000019198 oils Nutrition 0.000 description 15
- 229920001184 polypeptide Polymers 0.000 description 15
- PAIHPOGPJVUFJY-WDSKDSINSA-N Ala-Glu-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O PAIHPOGPJVUFJY-WDSKDSINSA-N 0.000 description 14
- QHASENCZLDHBGX-ONGXEEELSA-N Ala-Gly-Phe Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 QHASENCZLDHBGX-ONGXEEELSA-N 0.000 description 14
- OKEWAFFWMHBGPT-XPUUQOCRSA-N Ala-His-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](N)C)CC1=CN=CN1 OKEWAFFWMHBGPT-XPUUQOCRSA-N 0.000 description 14
- DCUCOIYYUBILPS-GUBZILKMSA-N Ala-Leu-Asp-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O DCUCOIYYUBILPS-GUBZILKMSA-N 0.000 description 14
- VCSABYLVNWQYQE-UHFFFAOYSA-N Ala-Lys-Lys Natural products NCCCCC(NC(=O)C(N)C)C(=O)NC(CCCCN)C(O)=O VCSABYLVNWQYQE-UHFFFAOYSA-N 0.000 description 14
- YCRAFFCYWOUEOF-DLOVCJGASA-N Ala-Phe-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C)CC1=CC=CC=C1 YCRAFFCYWOUEOF-DLOVCJGASA-N 0.000 description 14
- ARHJJAAWNWOACN-FXQIFTODSA-N Ala-Ser-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O ARHJJAAWNWOACN-FXQIFTODSA-N 0.000 description 14
- XMIAMUXIMWREBJ-HERUPUMHSA-N Ala-Trp-Asn Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CC(=O)N)C(=O)O)N XMIAMUXIMWREBJ-HERUPUMHSA-N 0.000 description 14
- FFEUXEAKYRCACT-PEDHHIEDSA-N Arg-Ile-Ile Chemical compound CC[C@H](C)[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CCCNC(N)=N)[C@@H](C)CC)C(O)=O FFEUXEAKYRCACT-PEDHHIEDSA-N 0.000 description 14
- JEXPNDORFYHJTM-IHRRRGAJSA-N Arg-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCCN=C(N)N JEXPNDORFYHJTM-IHRRRGAJSA-N 0.000 description 14
- HUAOKVVEVHACHR-CIUDSAMLSA-N Asn-Asp-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC(=O)N)N HUAOKVVEVHACHR-CIUDSAMLSA-N 0.000 description 14
- HPBNLFLSSQDFQW-WHFBIAKZSA-N Asn-Ser-Gly Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O HPBNLFLSSQDFQW-WHFBIAKZSA-N 0.000 description 14
- JPPLRQVZMZFOSX-UWJYBYFXSA-N Asn-Tyr-Ala Chemical compound NC(=O)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CC1=CC=C(O)C=C1 JPPLRQVZMZFOSX-UWJYBYFXSA-N 0.000 description 14
- HPNDBHLITCHRSO-WHFBIAKZSA-N Asp-Ala-Gly Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)NCC(O)=O HPNDBHLITCHRSO-WHFBIAKZSA-N 0.000 description 14
- KVMPVNGOKHTUHZ-GCJQMDKQSA-N Asp-Ala-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KVMPVNGOKHTUHZ-GCJQMDKQSA-N 0.000 description 14
- GWTLRDMPMJCNMH-WHFBIAKZSA-N Asp-Asn-Gly Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O GWTLRDMPMJCNMH-WHFBIAKZSA-N 0.000 description 14
- SNDBKTFJWVEVPO-WHFBIAKZSA-N Asp-Gly-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)NCC(=O)N[C@@H](CO)C(O)=O SNDBKTFJWVEVPO-WHFBIAKZSA-N 0.000 description 14
- BRRPVTUFESPTCP-ACZMJKKPSA-N Asp-Ser-Glu Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O BRRPVTUFESPTCP-ACZMJKKPSA-N 0.000 description 14
- YIDFBWRHIYOYAA-LKXGYXEUSA-N Asp-Ser-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O YIDFBWRHIYOYAA-LKXGYXEUSA-N 0.000 description 14
- KHNJVFYHIKLUPD-SRVKXCTJSA-N Gln-Leu-Met Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCSC)C(=O)O)NC(=O)[C@H](CCC(=O)N)N KHNJVFYHIKLUPD-SRVKXCTJSA-N 0.000 description 14
- KUBFPYIMAGXGBT-ACZMJKKPSA-N Gln-Ser-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O KUBFPYIMAGXGBT-ACZMJKKPSA-N 0.000 description 14
- RJONUNZIMUXUOI-GUBZILKMSA-N Glu-Asn-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CCC(=O)O)N RJONUNZIMUXUOI-GUBZILKMSA-N 0.000 description 14
- JPHYJQHPILOKHC-ACZMJKKPSA-N Glu-Asp-Asp Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O JPHYJQHPILOKHC-ACZMJKKPSA-N 0.000 description 14
- DWBBKNPKDHXIAC-SRVKXCTJSA-N Glu-Leu-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCC(O)=O DWBBKNPKDHXIAC-SRVKXCTJSA-N 0.000 description 14
- IDEODOAVGCMUQV-GUBZILKMSA-N Glu-Ser-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O IDEODOAVGCMUQV-GUBZILKMSA-N 0.000 description 14
- UMZHHILWZBFPGL-LOKLDPHHSA-N Glu-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)O)N)O UMZHHILWZBFPGL-LOKLDPHHSA-N 0.000 description 14
- HBMRTXJZQDVRFT-DZKIICNBSA-N Glu-Tyr-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C(C)C)C(O)=O HBMRTXJZQDVRFT-DZKIICNBSA-N 0.000 description 14
- UXJHNZODTMHWRD-WHFBIAKZSA-N Gly-Asn-Ala Chemical compound [H]NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(O)=O UXJHNZODTMHWRD-WHFBIAKZSA-N 0.000 description 14
- XEJTYSCIXKYSHR-WDSKDSINSA-N Gly-Asp-Gln Chemical compound C(CC(=O)N)[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)CN XEJTYSCIXKYSHR-WDSKDSINSA-N 0.000 description 14
- MHHUEAIBJZWDBH-YUMQZZPRSA-N Gly-Asp-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)CN MHHUEAIBJZWDBH-YUMQZZPRSA-N 0.000 description 14
- MOJKRXIRAZPZLW-WDSKDSINSA-N Gly-Glu-Ala Chemical compound [H]NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O MOJKRXIRAZPZLW-WDSKDSINSA-N 0.000 description 14
- LLZXNUUIBOALNY-QWRGUYRKSA-N Gly-Leu-Lys Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCCN LLZXNUUIBOALNY-QWRGUYRKSA-N 0.000 description 14
- VBOBNHSVQKKTOT-YUMQZZPRSA-N Gly-Lys-Ala Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O VBOBNHSVQKKTOT-YUMQZZPRSA-N 0.000 description 14
- UWQDKRIZSROAKS-FJXKBIBVSA-N Gly-Met-Thr Chemical compound [H]NCC(=O)N[C@@H](CCSC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O UWQDKRIZSROAKS-FJXKBIBVSA-N 0.000 description 14
- JYPCXBJRLBHWME-IUCAKERBSA-N Gly-Pro-Arg Chemical compound NCC(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(O)=O JYPCXBJRLBHWME-IUCAKERBSA-N 0.000 description 14
- JJGBXTYGTKWGAT-YUMQZZPRSA-N Gly-Pro-Glu Chemical compound NCC(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O JJGBXTYGTKWGAT-YUMQZZPRSA-N 0.000 description 14
- LYZYGGWCBLBDMC-QWHCGFSZSA-N Gly-Tyr-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=C(C=C2)O)NC(=O)CN)C(=O)O LYZYGGWCBLBDMC-QWHCGFSZSA-N 0.000 description 14
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 14
- RVKIPWVMZANZLI-UHFFFAOYSA-N H-Lys-Trp-OH Natural products C1=CC=C2C(CC(NC(=O)C(N)CCCCN)C(O)=O)=CNC2=C1 RVKIPWVMZANZLI-UHFFFAOYSA-N 0.000 description 14
- DFHVLUKTTVTCKY-PBCZWWQYSA-N His-Asn-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC1=CN=CN1)N)O DFHVLUKTTVTCKY-PBCZWWQYSA-N 0.000 description 14
- LBRCLQMZAHRTLV-ZKWXMUAHSA-N Ile-Gly-Ser Chemical compound CC[C@H](C)[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O LBRCLQMZAHRTLV-ZKWXMUAHSA-N 0.000 description 14
- QZZIBQZLWBOOJH-PEDHHIEDSA-N Ile-Ile-Val Chemical compound N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(=O)O QZZIBQZLWBOOJH-PEDHHIEDSA-N 0.000 description 14
- KCTIFOCXAIUQQK-QXEWZRGKSA-N Ile-Pro-Gly Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O KCTIFOCXAIUQQK-QXEWZRGKSA-N 0.000 description 14
- JZNVOBUNTWNZPW-GHCJXIJMSA-N Ile-Ser-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(=O)O)C(=O)O)N JZNVOBUNTWNZPW-GHCJXIJMSA-N 0.000 description 14
- GMUYXHHJAGQHGB-TUBUOCAGSA-N Ile-Thr-His Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N GMUYXHHJAGQHGB-TUBUOCAGSA-N 0.000 description 14
- SUPVSFFZWVOEOI-UHFFFAOYSA-N Leu-Ala-Tyr Natural products CC(C)CC(N)C(=O)NC(C)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 SUPVSFFZWVOEOI-UHFFFAOYSA-N 0.000 description 14
- BPANDPNDMJHFEV-CIUDSAMLSA-N Leu-Asp-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O BPANDPNDMJHFEV-CIUDSAMLSA-N 0.000 description 14
- UCDHVOALNXENLC-KBPBESRZSA-N Leu-Gly-Tyr Chemical compound CC(C)C[C@H]([NH3+])C(=O)NCC(=O)N[C@H](C([O-])=O)CC1=CC=C(O)C=C1 UCDHVOALNXENLC-KBPBESRZSA-N 0.000 description 14
- ZDBMWELMUCLUPL-QEJZJMRPSA-N Leu-Phe-Ala Chemical compound CC(C)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CC1=CC=CC=C1 ZDBMWELMUCLUPL-QEJZJMRPSA-N 0.000 description 14
- WMIOEVKKYIMVKI-DCAQKATOSA-N Leu-Pro-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O WMIOEVKKYIMVKI-DCAQKATOSA-N 0.000 description 14
- RGUXWMDNCPMQFB-YUMQZZPRSA-N Leu-Ser-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O RGUXWMDNCPMQFB-YUMQZZPRSA-N 0.000 description 14
- MVJRBCJCRYGCKV-GVXVVHGQSA-N Leu-Val-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O MVJRBCJCRYGCKV-GVXVVHGQSA-N 0.000 description 14
- FZIJIFCXUCZHOL-CIUDSAMLSA-N Lys-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN FZIJIFCXUCZHOL-CIUDSAMLSA-N 0.000 description 14
- BTSXLXFPMZXVPR-DLOVCJGASA-N Lys-Ala-His Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CCCCN)N BTSXLXFPMZXVPR-DLOVCJGASA-N 0.000 description 14
- FACUGMGEFUEBTI-SRVKXCTJSA-N Lys-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CCCCN FACUGMGEFUEBTI-SRVKXCTJSA-N 0.000 description 14
- NKKFVJRLCCUJNA-QWRGUYRKSA-N Lys-Gly-Lys Chemical compound NCCCC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCCN NKKFVJRLCCUJNA-QWRGUYRKSA-N 0.000 description 14
- MTBLFIQZECOEBY-IHRRRGAJSA-N Lys-Met-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(O)=O MTBLFIQZECOEBY-IHRRRGAJSA-N 0.000 description 14
- KSIPKXNIQOWMIC-RCWTZXSCSA-N Met-Thr-Arg Chemical compound CSCC[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CCCNC(N)=N KSIPKXNIQOWMIC-RCWTZXSCSA-N 0.000 description 14
- 108010002311 N-glycylglutamic acid Proteins 0.000 description 14
- SEPNOAFMZLLCEW-UBHSHLNASA-N Phe-Ala-Val Chemical compound N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)O SEPNOAFMZLLCEW-UBHSHLNASA-N 0.000 description 14
- NAXPHWZXEXNDIW-JTQLQIEISA-N Phe-Gly-Gly Chemical compound OC(=O)CNC(=O)CNC(=O)[C@@H](N)CC1=CC=CC=C1 NAXPHWZXEXNDIW-JTQLQIEISA-N 0.000 description 14
- KRYSMKKRRRWOCZ-QEWYBTABSA-N Phe-Ile-Glu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(O)=O KRYSMKKRRRWOCZ-QEWYBTABSA-N 0.000 description 14
- HFNPOYOKIPGAEI-SRVKXCTJSA-N Pro-Leu-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H]1CCCN1 HFNPOYOKIPGAEI-SRVKXCTJSA-N 0.000 description 14
- KIDXAAQVMNLJFQ-KZVJFYERSA-N Pro-Thr-Ala Chemical compound C[C@@H](O)[C@H](NC(=O)[C@@H]1CCCN1)C(=O)N[C@@H](C)C(O)=O KIDXAAQVMNLJFQ-KZVJFYERSA-N 0.000 description 14
- CDVFZMOFNJPUDD-ACZMJKKPSA-N Ser-Gln-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O CDVFZMOFNJPUDD-ACZMJKKPSA-N 0.000 description 14
- XKFJENWJGHMDLI-QWRGUYRKSA-N Ser-Phe-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)NCC(O)=O XKFJENWJGHMDLI-QWRGUYRKSA-N 0.000 description 14
- PYTKULIABVRXSC-BWBBJGPYSA-N Ser-Ser-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PYTKULIABVRXSC-BWBBJGPYSA-N 0.000 description 14
- FLMYSKVSDVHLEW-SVSWQMSJSA-N Ser-Thr-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O FLMYSKVSDVHLEW-SVSWQMSJSA-N 0.000 description 14
- DYEGLQRVMBWQLD-IXOXFDKPSA-N Ser-Thr-Phe Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H](CO)N)O DYEGLQRVMBWQLD-IXOXFDKPSA-N 0.000 description 14
- OJFFAQFRCVPHNN-JYBASQMISA-N Ser-Thr-Trp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O OJFFAQFRCVPHNN-JYBASQMISA-N 0.000 description 14
- OJRNZRROAIAHDL-LKXGYXEUSA-N Thr-Asn-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O OJRNZRROAIAHDL-LKXGYXEUSA-N 0.000 description 14
- HSQXHRIRJSFDOH-URLPEUOOSA-N Thr-Phe-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O HSQXHRIRJSFDOH-URLPEUOOSA-N 0.000 description 14
- HUPLKEHTTQBXSC-YJRXYDGGSA-N Thr-Ser-Tyr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 HUPLKEHTTQBXSC-YJRXYDGGSA-N 0.000 description 14
- BKVICMPZWRNWOC-RHYQMDGZSA-N Thr-Val-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)[C@@H](C)O BKVICMPZWRNWOC-RHYQMDGZSA-N 0.000 description 14
- DVWAIHZOPSYMSJ-ZVZYQTTQSA-N Trp-Glu-Val Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O)=CNC2=C1 DVWAIHZOPSYMSJ-ZVZYQTTQSA-N 0.000 description 14
- WLQRIHCMPFHGKP-PMVMPFDFSA-N Trp-Leu-Phe Chemical compound C([C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CC=1C2=CC=CC=C2NC=1)CC(C)C)C(O)=O)C1=CC=CC=C1 WLQRIHCMPFHGKP-PMVMPFDFSA-N 0.000 description 14
- LORJKYIPJIRIRT-BVSLBCMMSA-N Trp-Pro-Tyr Chemical compound C([C@H](NC(=O)[C@@H]1CCCN1C(=O)[C@H](CC=1C2=CC=CC=C2NC=1)N)C(O)=O)C1=CC=C(O)C=C1 LORJKYIPJIRIRT-BVSLBCMMSA-N 0.000 description 14
- ZWZOCUWOXSDYFZ-CQDKDKBSSA-N Tyr-Ala-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 ZWZOCUWOXSDYFZ-CQDKDKBSSA-N 0.000 description 14
- HSBZWINKRYZCSQ-KKUMJFAQSA-N Tyr-Lys-Asp Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(O)=O HSBZWINKRYZCSQ-KKUMJFAQSA-N 0.000 description 14
- OGPKMBOPMDTEDM-IHRRRGAJSA-N Tyr-Met-Ser Chemical compound CSCC[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N OGPKMBOPMDTEDM-IHRRRGAJSA-N 0.000 description 14
- WYOBRXPIZVKNMF-IRXDYDNUSA-N Tyr-Tyr-Gly Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)NCC(O)=O)C1=CC=C(O)C=C1 WYOBRXPIZVKNMF-IRXDYDNUSA-N 0.000 description 14
- FEFZWCSXEMVSPO-LSJOCFKGSA-N Val-His-Ala Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](Cc1cnc[nH]1)C(=O)N[C@@H](C)C(O)=O FEFZWCSXEMVSPO-LSJOCFKGSA-N 0.000 description 14
- WJVLTYSHNXRCLT-NHCYSSNCSA-N Val-His-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CC(=O)O)C(=O)O)N WJVLTYSHNXRCLT-NHCYSSNCSA-N 0.000 description 14
- WNZSAUMKZQXHNC-UKJIMTQDSA-N Val-Ile-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](C(C)C)N WNZSAUMKZQXHNC-UKJIMTQDSA-N 0.000 description 14
- LYERIXUFCYVFFX-GVXVVHGQSA-N Val-Leu-Glu Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](C(C)C)N LYERIXUFCYVFFX-GVXVVHGQSA-N 0.000 description 14
- ZRSZTKTVPNSUNA-IHRRRGAJSA-N Val-Lys-Leu Chemical compound CC(C)C[C@H](NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)C(C)C)C(O)=O ZRSZTKTVPNSUNA-IHRRRGAJSA-N 0.000 description 14
- VNGKMNPAENRGDC-JYJNAYRXSA-N Val-Phe-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)CC1=CC=CC=C1 VNGKMNPAENRGDC-JYJNAYRXSA-N 0.000 description 14
- 108010045350 alanyl-tyrosyl-alanine Proteins 0.000 description 14
- 108010069205 aspartyl-phenylalanine Proteins 0.000 description 14
- 108010092854 aspartyllysine Proteins 0.000 description 14
- 108010006664 gamma-glutamyl-glycyl-glycine Proteins 0.000 description 14
- HPAIKDPJURGQLN-UHFFFAOYSA-N glycyl-L-histidyl-L-phenylalanine Natural products C=1C=CC=CC=1CC(C(O)=O)NC(=O)C(NC(=O)CN)CC1=CN=CN1 HPAIKDPJURGQLN-UHFFFAOYSA-N 0.000 description 14
- 108010020688 glycylhistidine Proteins 0.000 description 14
- 108010081551 glycylphenylalanine Proteins 0.000 description 14
- 108010040030 histidinoalanine Proteins 0.000 description 14
- 108010090333 leucyl-lysyl-proline Proteins 0.000 description 14
- 108010009298 lysylglutamic acid Proteins 0.000 description 14
- 108010017391 lysylvaline Proteins 0.000 description 14
- 108010051242 phenylalanylserine Proteins 0.000 description 14
- 108010077112 prolyl-proline Proteins 0.000 description 14
- XSLGWYYNOSUMRM-ZKWXMUAHSA-N Ala-Val-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O XSLGWYYNOSUMRM-ZKWXMUAHSA-N 0.000 description 13
- DFCIPNHFKOQAME-FXQIFTODSA-N Arg-Ala-Asn Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(O)=O DFCIPNHFKOQAME-FXQIFTODSA-N 0.000 description 13
- KWKQGHSSNHPGOW-BQBZGAKWSA-N Arg-Ala-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)NCC(O)=O KWKQGHSSNHPGOW-BQBZGAKWSA-N 0.000 description 13
- YWEHYKGJWHPGPY-XGEHTFHBSA-N Cys-Thr-Arg Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](CS)N)O YWEHYKGJWHPGPY-XGEHTFHBSA-N 0.000 description 13
- 102000004190 Enzymes Human genes 0.000 description 13
- 108090000790 Enzymes Proteins 0.000 description 13
- XEYMBRRKIFYQMF-GUBZILKMSA-N Gln-Asp-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O XEYMBRRKIFYQMF-GUBZILKMSA-N 0.000 description 13
- JWNZHMSRZXXGTM-XKBZYTNZSA-N Glu-Ser-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O JWNZHMSRZXXGTM-XKBZYTNZSA-N 0.000 description 13
- 241000880493 Leptailurus serval Species 0.000 description 13
- RZXLZBIUTDQHJQ-SRVKXCTJSA-N Leu-Lys-Asp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(O)=O RZXLZBIUTDQHJQ-SRVKXCTJSA-N 0.000 description 13
- XYSXOCIWCPFOCG-IHRRRGAJSA-N Pro-Leu-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O XYSXOCIWCPFOCG-IHRRRGAJSA-N 0.000 description 13
- JFWDJFULOLKQFY-QWRGUYRKSA-N Ser-Gly-Phe Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O JFWDJFULOLKQFY-QWRGUYRKSA-N 0.000 description 13
- WAPFQMXRSDEGOE-IHRRRGAJSA-N Tyr-Glu-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O WAPFQMXRSDEGOE-IHRRRGAJSA-N 0.000 description 13
- 238000010276 construction Methods 0.000 description 13
- XQJAFSDFQZPYCU-UWJYBYFXSA-N Ala-Asn-Tyr Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N XQJAFSDFQZPYCU-UWJYBYFXSA-N 0.000 description 12
- BUDNAJYVCUHLSV-ZLUOBGJFSA-N Ala-Asp-Ser Chemical compound C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O BUDNAJYVCUHLSV-ZLUOBGJFSA-N 0.000 description 12
- LXAARTARZJJCMB-CIQUZCHMSA-N Ala-Ile-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LXAARTARZJJCMB-CIQUZCHMSA-N 0.000 description 12
- QUIGLPSHIFPEOV-CIUDSAMLSA-N Ala-Lys-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O QUIGLPSHIFPEOV-CIUDSAMLSA-N 0.000 description 12
- VCSABYLVNWQYQE-SRVKXCTJSA-N Ala-Lys-Lys Chemical compound NCCCC[C@H](NC(=O)[C@@H](N)C)C(=O)N[C@@H](CCCCN)C(O)=O VCSABYLVNWQYQE-SRVKXCTJSA-N 0.000 description 12
- ODLLXUGPBCQRPF-UHFFFAOYSA-N Ala-Met-Met-Tyr Chemical compound CSCCC(NC(=O)C(C)N)C(=O)NC(CCSC)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 ODLLXUGPBCQRPF-UHFFFAOYSA-N 0.000 description 12
- BTRULDJUUVGRNE-DCAQKATOSA-N Ala-Pro-Lys Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(O)=O BTRULDJUUVGRNE-DCAQKATOSA-N 0.000 description 12
- YFBGNGASPGRWEM-DCAQKATOSA-N Arg-Asp-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCCN=C(N)N)N YFBGNGASPGRWEM-DCAQKATOSA-N 0.000 description 12
- JZRLLSOWDYUKOK-SRVKXCTJSA-N Asn-Asp-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC(=O)N)N JZRLLSOWDYUKOK-SRVKXCTJSA-N 0.000 description 12
- NJSNXIOKBHPFMB-GMOBBJLQSA-N Asn-Pro-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](CC(=O)N)N NJSNXIOKBHPFMB-GMOBBJLQSA-N 0.000 description 12
- CBHVAFXKOYAHOY-NHCYSSNCSA-N Asn-Val-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O CBHVAFXKOYAHOY-NHCYSSNCSA-N 0.000 description 12
- JUWZKMBALYLZCK-WHFBIAKZSA-N Asp-Gly-Asn Chemical compound OC(=O)C[C@H](N)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O JUWZKMBALYLZCK-WHFBIAKZSA-N 0.000 description 12
- RQHLMGCXCZUOGT-ZPFDUUQYSA-N Asp-Leu-Ile Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O RQHLMGCXCZUOGT-ZPFDUUQYSA-N 0.000 description 12
- UJGRZQYSNYTCAX-SRVKXCTJSA-N Asp-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(O)=O UJGRZQYSNYTCAX-SRVKXCTJSA-N 0.000 description 12
- GYWQGGUCMDCUJE-DLOVCJGASA-N Asp-Phe-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C)C(O)=O GYWQGGUCMDCUJE-DLOVCJGASA-N 0.000 description 12
- SQIARYGNVQWOSB-BZSNNMDCSA-N Asp-Tyr-Phe Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O SQIARYGNVQWOSB-BZSNNMDCSA-N 0.000 description 12
- 101100512078 Caenorhabditis elegans lys-1 gene Proteins 0.000 description 12
- NROSLUJMIQGFKS-IUCAKERBSA-N Gln-His-Gly Chemical compound C1=C(NC=N1)C[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CCC(=O)N)N NROSLUJMIQGFKS-IUCAKERBSA-N 0.000 description 12
- XZUUUKNKNWVPHQ-JYJNAYRXSA-N Gln-Phe-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O XZUUUKNKNWVPHQ-JYJNAYRXSA-N 0.000 description 12
- PXXGVUVQWQGGIG-YUMQZZPRSA-N Glu-Gly-Arg Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCN=C(N)N PXXGVUVQWQGGIG-YUMQZZPRSA-N 0.000 description 12
- ITBHUUMCJJQUSC-LAEOZQHASA-N Glu-Ile-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(O)=O ITBHUUMCJJQUSC-LAEOZQHASA-N 0.000 description 12
- BGVYNAQWHSTTSP-BYULHYEWSA-N Gly-Asn-Ile Chemical compound [H]NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O BGVYNAQWHSTTSP-BYULHYEWSA-N 0.000 description 12
- DBUNZBWUWCIELX-JHEQGTHGSA-N Gly-Thr-Glu Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(O)=O DBUNZBWUWCIELX-JHEQGTHGSA-N 0.000 description 12
- ZVXMEWXHFBYJPI-LSJOCFKGSA-N Gly-Val-Ile Chemical compound [H]NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O ZVXMEWXHFBYJPI-LSJOCFKGSA-N 0.000 description 12
- QICVAHODWHIWIS-HTFCKZLJSA-N Ile-Ala-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N QICVAHODWHIWIS-HTFCKZLJSA-N 0.000 description 12
- BGZIJZJBXRVBGJ-SXTJYALSSA-N Ile-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N BGZIJZJBXRVBGJ-SXTJYALSSA-N 0.000 description 12
- UAQSZXGJGLHMNV-XEGUGMAKSA-N Ile-Gly-Tyr Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N UAQSZXGJGLHMNV-XEGUGMAKSA-N 0.000 description 12
- YCKPUHHMCFSUMD-IUKAMOBKSA-N Ile-Thr-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(=O)O)C(=O)O)N YCKPUHHMCFSUMD-IUKAMOBKSA-N 0.000 description 12
- SUPVSFFZWVOEOI-CQDKDKBSSA-N Leu-Ala-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 SUPVSFFZWVOEOI-CQDKDKBSSA-N 0.000 description 12
- GLBNEGIOFRVRHO-JYJNAYRXSA-N Leu-Gln-Phe Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O GLBNEGIOFRVRHO-JYJNAYRXSA-N 0.000 description 12
- VGPCJSXPPOQPBK-YUMQZZPRSA-N Leu-Gly-Ser Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O VGPCJSXPPOQPBK-YUMQZZPRSA-N 0.000 description 12
- VCHVSKNMTXWIIP-SRVKXCTJSA-N Leu-Lys-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O VCHVSKNMTXWIIP-SRVKXCTJSA-N 0.000 description 12
- CHJKEDSZNSONPS-DCAQKATOSA-N Leu-Pro-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O CHJKEDSZNSONPS-DCAQKATOSA-N 0.000 description 12
- BGGTYDNTOYRTTR-MEYUZBJRSA-N Leu-Tyr-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](CC(C)C)N)O BGGTYDNTOYRTTR-MEYUZBJRSA-N 0.000 description 12
- XFIHDSBIPWEYJJ-YUMQZZPRSA-N Lys-Ala-Gly Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN XFIHDSBIPWEYJJ-YUMQZZPRSA-N 0.000 description 12
- GKFNXYMAMKJSKD-NHCYSSNCSA-N Lys-Asp-Val Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O GKFNXYMAMKJSKD-NHCYSSNCSA-N 0.000 description 12
- DTUZCYRNEJDKSR-NHCYSSNCSA-N Lys-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN DTUZCYRNEJDKSR-NHCYSSNCSA-N 0.000 description 12
- XDPLZVNMYQOFQZ-BJDJZHNGSA-N Lys-Ile-Cys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CCCCN)N XDPLZVNMYQOFQZ-BJDJZHNGSA-N 0.000 description 12
- RBEATVHTWHTHTJ-KKUMJFAQSA-N Lys-Leu-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O RBEATVHTWHTHTJ-KKUMJFAQSA-N 0.000 description 12
- WINFHLHJTRGLCV-BZSNNMDCSA-N Lys-Tyr-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CCCCN)C(O)=O)CC1=CC=C(O)C=C1 WINFHLHJTRGLCV-BZSNNMDCSA-N 0.000 description 12
- LBSWWNKMVPAXOI-GUBZILKMSA-N Met-Val-Ser Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O LBSWWNKMVPAXOI-GUBZILKMSA-N 0.000 description 12
- HCTXJGRYAACKOB-SRVKXCTJSA-N Phe-Asn-Asp Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)N HCTXJGRYAACKOB-SRVKXCTJSA-N 0.000 description 12
- MSHZERMPZKCODG-ACRUOGEOSA-N Phe-Leu-Phe Chemical compound C([C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 MSHZERMPZKCODG-ACRUOGEOSA-N 0.000 description 12
- FGWUALWGCZJQDJ-URLPEUOOSA-N Phe-Thr-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O FGWUALWGCZJQDJ-URLPEUOOSA-N 0.000 description 12
- OOLOTUZJUBOMAX-GUBZILKMSA-N Pro-Ala-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O OOLOTUZJUBOMAX-GUBZILKMSA-N 0.000 description 12
- QUBVFEANYYWBTM-VEVYYDQMSA-N Pro-Thr-Asp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(O)=O QUBVFEANYYWBTM-VEVYYDQMSA-N 0.000 description 12
- GHPQVUYZQQGEDA-BIIVOSGPSA-N Ser-Asp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)O)NC(=O)[C@H](CO)N)C(=O)O GHPQVUYZQQGEDA-BIIVOSGPSA-N 0.000 description 12
- IFPBAGJBHSNYPR-ZKWXMUAHSA-N Ser-Ile-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(O)=O IFPBAGJBHSNYPR-ZKWXMUAHSA-N 0.000 description 12
- NLSNVZAREYQMGR-HJGDQZAQSA-N Thr-Asp-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O NLSNVZAREYQMGR-HJGDQZAQSA-N 0.000 description 12
- DJDSEDOKJTZBAR-ZDLURKLDSA-N Thr-Gly-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O DJDSEDOKJTZBAR-ZDLURKLDSA-N 0.000 description 12
- ADPHPKGWVDHWML-PPCPHDFISA-N Thr-Ile-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)O)NC(=O)[C@H]([C@@H](C)O)N ADPHPKGWVDHWML-PPCPHDFISA-N 0.000 description 12
- HOVLHEKTGVIKAP-WDCWCFNPSA-N Thr-Leu-Gln Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O HOVLHEKTGVIKAP-WDCWCFNPSA-N 0.000 description 12
- ISLDRLHVPXABBC-IEGACIPQSA-N Thr-Leu-Trp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O ISLDRLHVPXABBC-IEGACIPQSA-N 0.000 description 12
- OGOYMQWIWHGTGH-KZVJFYERSA-N Thr-Val-Ala Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O OGOYMQWIWHGTGH-KZVJFYERSA-N 0.000 description 12
- ARPONUQDNWLXOZ-KKUMJFAQSA-N Tyr-Gln-Arg Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O ARPONUQDNWLXOZ-KKUMJFAQSA-N 0.000 description 12
- ISERLACIZUGCDX-ZKWXMUAHSA-N Val-Asp-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](C(C)C)N ISERLACIZUGCDX-ZKWXMUAHSA-N 0.000 description 12
- FEXILLGKGGTLRI-NHCYSSNCSA-N Val-Leu-Asn Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](C(C)C)N FEXILLGKGGTLRI-NHCYSSNCSA-N 0.000 description 12
- KRAHMIJVUPUOTQ-DCAQKATOSA-N Val-Ser-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N KRAHMIJVUPUOTQ-DCAQKATOSA-N 0.000 description 12
- YQYFYUSYEDNLSD-YEPSODPASA-N Val-Thr-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O YQYFYUSYEDNLSD-YEPSODPASA-N 0.000 description 12
- LCHZBEUVGAVMKS-RHYQMDGZSA-N Val-Thr-Leu Chemical compound CC(C)C[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)[C@@H](C)O)C(O)=O LCHZBEUVGAVMKS-RHYQMDGZSA-N 0.000 description 12
- QHSSPPHOHJSTML-HOCLYGCPSA-N Val-Trp-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)NCC(=O)O)N QHSSPPHOHJSTML-HOCLYGCPSA-N 0.000 description 12
- 108010025307 buforin II Proteins 0.000 description 12
- 239000003925 fat Substances 0.000 description 12
- 235000019197 fats Nutrition 0.000 description 12
- 108010049041 glutamylalanine Proteins 0.000 description 12
- 108010028188 glycyl-histidyl-serine Proteins 0.000 description 12
- 108010077515 glycylproline Proteins 0.000 description 12
- 230000002366 lipolytic effect Effects 0.000 description 12
- 102000004169 proteins and genes Human genes 0.000 description 12
- LGFCAXJBAZESCF-ACZMJKKPSA-N Ala-Gln-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O LGFCAXJBAZESCF-ACZMJKKPSA-N 0.000 description 11
- VBRDBGCROKWTPV-XHNCKOQMSA-N Ala-Glu-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N1CCC[C@@H]1C(=O)O)N VBRDBGCROKWTPV-XHNCKOQMSA-N 0.000 description 11
- SIGTYDNEPYEXGK-ZANVPECISA-N Ala-Gly-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)CNC(=O)[C@@H](N)C)C(O)=O)=CNC2=C1 SIGTYDNEPYEXGK-ZANVPECISA-N 0.000 description 11
- GKAZXNDATBWNBI-DCAQKATOSA-N Ala-Met-Lys Chemical compound C[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(=O)O)N GKAZXNDATBWNBI-DCAQKATOSA-N 0.000 description 11
- XHFXZQHTLJVZBN-FXQIFTODSA-N Asn-Arg-Asn Chemical compound C(C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N)CN=C(N)N XHFXZQHTLJVZBN-FXQIFTODSA-N 0.000 description 11
- UMHUHHJMEXNSIV-CIUDSAMLSA-N Asp-Leu-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(O)=O UMHUHHJMEXNSIV-CIUDSAMLSA-N 0.000 description 11
- ITYRYNUZHPNCIK-GUBZILKMSA-N Glu-Ala-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O ITYRYNUZHPNCIK-GUBZILKMSA-N 0.000 description 11
- VIPDPMHGICREIS-GVXVVHGQSA-N Glu-Val-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O VIPDPMHGICREIS-GVXVVHGQSA-N 0.000 description 11
- LQSBBHNVAVNZSX-GHCJXIJMSA-N Ile-Ala-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](CC(=O)N)C(=O)O)N LQSBBHNVAVNZSX-GHCJXIJMSA-N 0.000 description 11
- ODPKZZLRDNXTJZ-WHOFXGATSA-N Ile-Gly-Phe Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N ODPKZZLRDNXTJZ-WHOFXGATSA-N 0.000 description 11
- SEZADXQOJJTXPG-VFAJRCTISA-N Lys-Thr-Trp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](CCCCN)N)O SEZADXQOJJTXPG-VFAJRCTISA-N 0.000 description 11
- FUOGXAQMNJMBFG-WPRPVWTQSA-N Pro-Val-Gly Chemical compound OC(=O)CNC(=O)[C@H](C(C)C)NC(=O)[C@@H]1CCCN1 FUOGXAQMNJMBFG-WPRPVWTQSA-N 0.000 description 11
- NLOAIFSWUUFQFR-CIUDSAMLSA-N Ser-Leu-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O NLOAIFSWUUFQFR-CIUDSAMLSA-N 0.000 description 11
- UGGWCAFQPKANMW-FXQIFTODSA-N Ser-Met-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](C)C(O)=O UGGWCAFQPKANMW-FXQIFTODSA-N 0.000 description 11
- MFEBUIFJVPNZLO-OLHMAJIHSA-N Thr-Asp-Asn Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O MFEBUIFJVPNZLO-OLHMAJIHSA-N 0.000 description 11
- VTMGKRABARCZAX-OSUNSFLBSA-N Thr-Pro-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)[C@@H](C)O VTMGKRABARCZAX-OSUNSFLBSA-N 0.000 description 11
- KLQPIEVIKOQRAW-IZPVPAKOSA-N Tyr-Thr-Thr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N)O KLQPIEVIKOQRAW-IZPVPAKOSA-N 0.000 description 11
- 239000000243 solution Substances 0.000 description 11
- 108020001019 DNA Primers Proteins 0.000 description 10
- 239000003155 DNA primer Substances 0.000 description 10
- CCBIBMKQNXHNIN-ZETCQYMHSA-N Gly-Leu-Gly Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O CCBIBMKQNXHNIN-ZETCQYMHSA-N 0.000 description 10
- BAYQNCWLXIDLHX-ONGXEEELSA-N Gly-Val-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)CN BAYQNCWLXIDLHX-ONGXEEELSA-N 0.000 description 10
- TZCGZYWNIDZZMR-UHFFFAOYSA-N Ile-Arg-Ala Natural products CCC(C)C(N)C(=O)NC(C(=O)NC(C)C(O)=O)CCCN=C(N)N TZCGZYWNIDZZMR-UHFFFAOYSA-N 0.000 description 10
- BQVUABVGYYSDCJ-UHFFFAOYSA-N Nalpha-L-Leucyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)CC(C)C)C(O)=O)=CNC2=C1 BQVUABVGYYSDCJ-UHFFFAOYSA-N 0.000 description 10
- OHAJHDJOCKKJLV-LKXGYXEUSA-N Thr-Asp-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O OHAJHDJOCKKJLV-LKXGYXEUSA-N 0.000 description 10
- UIRPULWLRODAEQ-QEJZJMRPSA-N Trp-Ser-Glu Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(O)=O)=CNC2=C1 UIRPULWLRODAEQ-QEJZJMRPSA-N 0.000 description 10
- DLZKEQQWXODGGZ-KWQFWETISA-N Tyr-Ala-Gly Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 DLZKEQQWXODGGZ-KWQFWETISA-N 0.000 description 10
- UZFNHAXYMICTBU-DZKIICNBSA-N Val-Phe-Gln Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N UZFNHAXYMICTBU-DZKIICNBSA-N 0.000 description 10
- 108010072405 glycyl-aspartyl-glycine Proteins 0.000 description 10
- 108010054666 glycyl-leucyl-glycyl-glycine Proteins 0.000 description 10
- LVZSQWIWCANHPF-UHFFFAOYSA-N p-nitrophenyl palmitate Chemical compound CCCCCCCCCCCCCCCC(=O)OC1=CC=C([N+]([O-])=O)C=C1 LVZSQWIWCANHPF-UHFFFAOYSA-N 0.000 description 10
- 108010084525 phenylalanyl-phenylalanyl-glycine Proteins 0.000 description 10
- 235000018102 proteins Nutrition 0.000 description 10
- 108010072986 threonyl-seryl-lysine Proteins 0.000 description 10
- XBQSLMACWDXWLJ-GHCJXIJMSA-N Asp-Ala-Ile Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O XBQSLMACWDXWLJ-GHCJXIJMSA-N 0.000 description 9
- SPKCGKRUYKMDHP-GUDRVLHUSA-N Asp-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC(=O)O)N SPKCGKRUYKMDHP-GUDRVLHUSA-N 0.000 description 9
- ICYRCNICGBJLGM-HJGDQZAQSA-N Leu-Thr-Asp Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(O)=O ICYRCNICGBJLGM-HJGDQZAQSA-N 0.000 description 9
- JKXVPNCSAMWUEJ-GUBZILKMSA-N Met-Met-Asp Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(O)=O JKXVPNCSAMWUEJ-GUBZILKMSA-N 0.000 description 9
- QARCDOCCDOLJSF-HJPIBITLSA-N Tyr-Ile-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N QARCDOCCDOLJSF-HJPIBITLSA-N 0.000 description 9
- QZKVWWIUSQGWMY-IHRRRGAJSA-N Val-Ser-Phe Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 QZKVWWIUSQGWMY-IHRRRGAJSA-N 0.000 description 9
- 235000019441 ethanol Nutrition 0.000 description 9
- 239000002699 waste material Substances 0.000 description 9
- HXNNRBHASOSVPG-GUBZILKMSA-N Ala-Glu-Leu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O HXNNRBHASOSVPG-GUBZILKMSA-N 0.000 description 8
- OINVDEKBKBCPLX-JXUBOQSCSA-N Ala-Lys-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OINVDEKBKBCPLX-JXUBOQSCSA-N 0.000 description 8
- DCVYRWFAMZFSDA-ZLUOBGJFSA-N Ala-Ser-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O DCVYRWFAMZFSDA-ZLUOBGJFSA-N 0.000 description 8
- VBFJESQBIWCWRL-DCAQKATOSA-N Arg-Ala-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCNC(N)=N VBFJESQBIWCWRL-DCAQKATOSA-N 0.000 description 8
- GMFAGHNRXPSSJS-SRVKXCTJSA-N Arg-Leu-Gln Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O GMFAGHNRXPSSJS-SRVKXCTJSA-N 0.000 description 8
- ZDOQDYFZNGASEY-BIIVOSGPSA-N Asn-Asp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC(=O)N)N)C(=O)O ZDOQDYFZNGASEY-BIIVOSGPSA-N 0.000 description 8
- FTNVLGCFIJEMQT-CIUDSAMLSA-N Asp-Cys-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CC(=O)O)N FTNVLGCFIJEMQT-CIUDSAMLSA-N 0.000 description 8
- 241000588724 Escherichia coli Species 0.000 description 8
- YKBUCXNNBYZYAY-MNXVOIDGSA-N Glu-Lys-Ile Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O YKBUCXNNBYZYAY-MNXVOIDGSA-N 0.000 description 8
- JMQFHZWESBGPFC-WDSKDSINSA-N Gly-Gln-Asp Chemical compound NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O JMQFHZWESBGPFC-WDSKDSINSA-N 0.000 description 8
- LPCKHUXOGVNZRS-YUMQZZPRSA-N Gly-His-Ser Chemical compound [H]NCC(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CO)C(O)=O LPCKHUXOGVNZRS-YUMQZZPRSA-N 0.000 description 8
- FFALDIDGPLUDKV-ZDLURKLDSA-N Gly-Thr-Ser Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O FFALDIDGPLUDKV-ZDLURKLDSA-N 0.000 description 8
- RYAOJUMWLWUGNW-QMMMGPOBSA-N Gly-Val-Gly Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O RYAOJUMWLWUGNW-QMMMGPOBSA-N 0.000 description 8
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 8
- JWTKVPMQCCRPQY-SRVKXCTJSA-N His-Asn-Leu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O JWTKVPMQCCRPQY-SRVKXCTJSA-N 0.000 description 8
- SACHLUOUHCVIKI-GMOBBJLQSA-N Ile-Arg-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)N SACHLUOUHCVIKI-GMOBBJLQSA-N 0.000 description 8
- JXMSHKFPDIUYGS-SIUGBPQLSA-N Ile-Glu-Tyr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N JXMSHKFPDIUYGS-SIUGBPQLSA-N 0.000 description 8
- UIEZQYNXCYHMQS-BJDJZHNGSA-N Ile-Lys-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)O)N UIEZQYNXCYHMQS-BJDJZHNGSA-N 0.000 description 8
- SAVXZJYTTQQQDD-QEWYBTABSA-N Ile-Phe-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N SAVXZJYTTQQQDD-QEWYBTABSA-N 0.000 description 8
- HYIFFZAQXPUEAU-QWRGUYRKSA-N Leu-Gly-Leu Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC(C)C HYIFFZAQXPUEAU-QWRGUYRKSA-N 0.000 description 8
- LKDXINHHSWFFJC-SRVKXCTJSA-N Lys-Ser-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CCCCN)N LKDXINHHSWFFJC-SRVKXCTJSA-N 0.000 description 8
- RMKJOQSYLQQRFN-KKUMJFAQSA-N Lys-Tyr-Asp Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(O)=O)C(O)=O RMKJOQSYLQQRFN-KKUMJFAQSA-N 0.000 description 8
- HAQLBBVZAGMESV-IHRRRGAJSA-N Met-Lys-Lys Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(O)=O HAQLBBVZAGMESV-IHRRRGAJSA-N 0.000 description 8
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 8
- AUEJLPRZGVVDNU-UHFFFAOYSA-N N-L-tyrosyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CC1=CC=C(O)C=C1 AUEJLPRZGVVDNU-UHFFFAOYSA-N 0.000 description 8
- FPTXMUIBLMGTQH-ONGXEEELSA-N Phe-Ala-Gly Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=CC=C1 FPTXMUIBLMGTQH-ONGXEEELSA-N 0.000 description 8
- JXQVYPWVGUOIDV-MXAVVETBSA-N Phe-Ser-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JXQVYPWVGUOIDV-MXAVVETBSA-N 0.000 description 8
- MLKVIVZCFYRTIR-KKUMJFAQSA-N Pro-Phe-Gln Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(N)=O)C(O)=O MLKVIVZCFYRTIR-KKUMJFAQSA-N 0.000 description 8
- FIODMZKLZFLYQP-GUBZILKMSA-N Pro-Val-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O FIODMZKLZFLYQP-GUBZILKMSA-N 0.000 description 8
- VQBCMLMPEWPUTB-ACZMJKKPSA-N Ser-Glu-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O VQBCMLMPEWPUTB-ACZMJKKPSA-N 0.000 description 8
- OWCVUSJMEBGMOK-YUMQZZPRSA-N Ser-Lys-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O OWCVUSJMEBGMOK-YUMQZZPRSA-N 0.000 description 8
- LRZLZIUXQBIWTB-KATARQTJSA-N Ser-Lys-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LRZLZIUXQBIWTB-KATARQTJSA-N 0.000 description 8
- LPSKHZWBQONOQJ-XIRDDKMYSA-N Ser-Lys-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)N LPSKHZWBQONOQJ-XIRDDKMYSA-N 0.000 description 8
- FLONGDPORFIVQW-XGEHTFHBSA-N Ser-Pro-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CO FLONGDPORFIVQW-XGEHTFHBSA-N 0.000 description 8
- XFTYVCHLARBHBQ-FOHZUACHSA-N Thr-Gly-Asn Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O XFTYVCHLARBHBQ-FOHZUACHSA-N 0.000 description 8
- AKLNEFNQWLHIGY-QWRGUYRKSA-N Tyr-Gly-Asp Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)NCC(=O)N[C@@H](CC(=O)O)C(=O)O)N)O AKLNEFNQWLHIGY-QWRGUYRKSA-N 0.000 description 8
- QWCZXKIFPWPQHR-JYJNAYRXSA-N Val-Pro-Tyr Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 QWCZXKIFPWPQHR-JYJNAYRXSA-N 0.000 description 8
- 108010070643 prolylglutamic acid Proteins 0.000 description 8
- 108010090894 prolylleucine Proteins 0.000 description 8
- 108010078580 tyrosylleucine Proteins 0.000 description 8
- FRBAHXABMQXSJQ-FXQIFTODSA-N Arg-Ser-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O FRBAHXABMQXSJQ-FXQIFTODSA-N 0.000 description 7
- LEFKSBYHUGUWLP-ACZMJKKPSA-N Asn-Ala-Glu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O LEFKSBYHUGUWLP-ACZMJKKPSA-N 0.000 description 7
- TVVYVAUGRHNTGT-UGYAYLCHSA-N Asp-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC(O)=O TVVYVAUGRHNTGT-UGYAYLCHSA-N 0.000 description 7
- RUDRIZRGOLQSMX-IUCAKERBSA-N Gly-Met-Met Chemical compound [H]NCC(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCSC)C(O)=O RUDRIZRGOLQSMX-IUCAKERBSA-N 0.000 description 7
- PZAJPILZRFPYJJ-SRVKXCTJSA-N His-Ser-Leu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O PZAJPILZRFPYJJ-SRVKXCTJSA-N 0.000 description 7
- ZNOBVZFCHNHKHA-KBIXCLLPSA-N Ile-Ser-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N ZNOBVZFCHNHKHA-KBIXCLLPSA-N 0.000 description 7
- FQZPTCNSNPWHLJ-AVGNSLFASA-N Leu-Gln-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(O)=O FQZPTCNSNPWHLJ-AVGNSLFASA-N 0.000 description 7
- OVZLLFONXILPDZ-VOAKCMCISA-N Leu-Lys-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OVZLLFONXILPDZ-VOAKCMCISA-N 0.000 description 7
- ONPDTSFZAIWMDI-AVGNSLFASA-N Lys-Leu-Gln Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O ONPDTSFZAIWMDI-AVGNSLFASA-N 0.000 description 7
- VTFXTWDFPTWNJY-RHYQMDGZSA-N Pro-Leu-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O VTFXTWDFPTWNJY-RHYQMDGZSA-N 0.000 description 7
- 108010093581 aspartyl-proline Proteins 0.000 description 7
- 108010005942 methionylglycine Proteins 0.000 description 7
- 108010071207 serylmethionine Proteins 0.000 description 7
- GJLXVWOMRRWCIB-MERZOTPQSA-N (2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-acetamido-5-(diaminomethylideneamino)pentanoyl]amino]-3-(4-hydroxyphenyl)propanoyl]amino]-3-(4-hydroxyphenyl)propanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]amino]-3-(1H-indol-3-yl)propanoyl]amino]-6-aminohexanoyl]amino]-6-aminohexanoyl]amino]-6-aminohexanoyl]amino]-6-aminohexanoyl]amino]-6-aminohexanoyl]amino]-6-aminohexanoyl]amino]-6-aminohexanamide Chemical compound C([C@H](NC(=O)[C@H](CCCN=C(N)N)NC(=O)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(N)=O)C1=CC=C(O)C=C1 GJLXVWOMRRWCIB-MERZOTPQSA-N 0.000 description 6
- MMLHRUJLOUSRJX-CIUDSAMLSA-N Ala-Ser-Lys Chemical compound C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCCN MMLHRUJLOUSRJX-CIUDSAMLSA-N 0.000 description 6
- NZGRHTKZFSVPAN-BIIVOSGPSA-N Ala-Ser-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CO)C(=O)N1CCC[C@@H]1C(=O)O)N NZGRHTKZFSVPAN-BIIVOSGPSA-N 0.000 description 6
- PGNNQOJOEGFAOR-KWQFWETISA-N Ala-Tyr-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](N)C)CC1=CC=C(O)C=C1 PGNNQOJOEGFAOR-KWQFWETISA-N 0.000 description 6
- OLGCWMNDJTWQAG-GUBZILKMSA-N Asn-Glu-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC(N)=O OLGCWMNDJTWQAG-GUBZILKMSA-N 0.000 description 6
- VHQSGALUSWIYOD-QXEWZRGKSA-N Asn-Pro-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O VHQSGALUSWIYOD-QXEWZRGKSA-N 0.000 description 6
- AKPLMZMNJGNUKT-ZLUOBGJFSA-N Asp-Asp-Cys Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CS)C(O)=O AKPLMZMNJGNUKT-ZLUOBGJFSA-N 0.000 description 6
- PYXXJFRXIYAESU-PCBIJLKTSA-N Asp-Ile-Phe Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O PYXXJFRXIYAESU-PCBIJLKTSA-N 0.000 description 6
- CZECQDPEMSVPDH-MNXVOIDGSA-N Asp-Leu-Val-Ser Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O CZECQDPEMSVPDH-MNXVOIDGSA-N 0.000 description 6
- ZVGRHIRJLWBWGJ-ACZMJKKPSA-N Asp-Ser-Gln Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(O)=O ZVGRHIRJLWBWGJ-ACZMJKKPSA-N 0.000 description 6
- PLNJUJGNLDSFOP-UWJYBYFXSA-N Asp-Tyr-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C)C(O)=O PLNJUJGNLDSFOP-UWJYBYFXSA-N 0.000 description 6
- VSXBYIJUAXPAAL-WDSKDSINSA-N Gln-Gly-Ala Chemical compound OC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H](N)CCC(N)=O VSXBYIJUAXPAAL-WDSKDSINSA-N 0.000 description 6
- MTCXQQINVAFZKW-MNXVOIDGSA-N Gln-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)N)N MTCXQQINVAFZKW-MNXVOIDGSA-N 0.000 description 6
- YKLNMGJYMNPBCP-ACZMJKKPSA-N Glu-Asn-Asp Chemical compound C(CC(=O)O)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)N YKLNMGJYMNPBCP-ACZMJKKPSA-N 0.000 description 6
- HILMIYALTUQTRC-XVKPBYJWSA-N Glu-Gly-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O HILMIYALTUQTRC-XVKPBYJWSA-N 0.000 description 6
- PDUHNKAFQXQNLH-ZETCQYMHSA-N Gly-Lys-Gly Chemical compound NCCCC[C@H](NC(=O)CN)C(=O)NCC(O)=O PDUHNKAFQXQNLH-ZETCQYMHSA-N 0.000 description 6
- CVFOYJJOZYYEPE-KBPBESRZSA-N Gly-Lys-Tyr Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O CVFOYJJOZYYEPE-KBPBESRZSA-N 0.000 description 6
- UOYGZBIPZYKGSH-SRVKXCTJSA-N His-Ser-Lys Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)O)N UOYGZBIPZYKGSH-SRVKXCTJSA-N 0.000 description 6
- REXAUQBGSGDEJY-IGISWZIWSA-N Ile-Tyr-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N REXAUQBGSGDEJY-IGISWZIWSA-N 0.000 description 6
- PWWVAXIEGOYWEE-UHFFFAOYSA-N Isophenergan Chemical compound C1=CC=C2N(CC(C)N(C)C)C3=CC=CC=C3SC2=C1 PWWVAXIEGOYWEE-UHFFFAOYSA-N 0.000 description 6
- VWHGTYCRDRBSFI-ZETCQYMHSA-N Leu-Gly-Gly Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)NCC(O)=O VWHGTYCRDRBSFI-ZETCQYMHSA-N 0.000 description 6
- HGFGEMSVBMCFKK-MNXVOIDGSA-N Leu-Ile-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(O)=O HGFGEMSVBMCFKK-MNXVOIDGSA-N 0.000 description 6
- HRTRLSRYZZKPCO-BJDJZHNGSA-N Leu-Ile-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O HRTRLSRYZZKPCO-BJDJZHNGSA-N 0.000 description 6
- WBRJVRXEGQIDRK-XIRDDKMYSA-N Leu-Trp-Ser Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](N)CC(C)C)C(=O)N[C@@H](CO)C(O)=O)=CNC2=C1 WBRJVRXEGQIDRK-XIRDDKMYSA-N 0.000 description 6
- GQZMPWBZQALKJO-UWVGGRQHSA-N Lys-Gly-Arg Chemical compound [H]N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O GQZMPWBZQALKJO-UWVGGRQHSA-N 0.000 description 6
- LSXGADJXBDFXQU-DLOVCJGASA-N Phe-Ala-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=CC=C1 LSXGADJXBDFXQU-DLOVCJGASA-N 0.000 description 6
- ZLGQEBCCANLYRA-RYUDHWBXSA-N Phe-Gly-Glu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O ZLGQEBCCANLYRA-RYUDHWBXSA-N 0.000 description 6
- IWZRODDWOSIXPZ-IRXDYDNUSA-N Phe-Phe-Gly Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)NCC(O)=O)C1=CC=CC=C1 IWZRODDWOSIXPZ-IRXDYDNUSA-N 0.000 description 6
- MHHQQZIFLWFZGR-DCAQKATOSA-N Pro-Lys-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O MHHQQZIFLWFZGR-DCAQKATOSA-N 0.000 description 6
- ZUZINZIJHJFJRN-UBHSHLNASA-N Pro-Phe-Ala Chemical compound C([C@@H](C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@H]1NCCC1)C1=CC=CC=C1 ZUZINZIJHJFJRN-UBHSHLNASA-N 0.000 description 6
- GFHXZNVJIKMAGO-IHRRRGAJSA-N Pro-Phe-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O GFHXZNVJIKMAGO-IHRRRGAJSA-N 0.000 description 6
- YQHZVYJAGWMHES-ZLUOBGJFSA-N Ser-Ala-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O YQHZVYJAGWMHES-ZLUOBGJFSA-N 0.000 description 6
- WPAKPLPGQNUXGN-OSUNSFLBSA-N Thr-Ile-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O WPAKPLPGQNUXGN-OSUNSFLBSA-N 0.000 description 6
- SIEZEMFJLYRUMK-YTWAJWBKSA-N Thr-Met-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCSC)C(=O)N1CCC[C@@H]1C(=O)O)N)O SIEZEMFJLYRUMK-YTWAJWBKSA-N 0.000 description 6
- IQPWNQRRAJHOKV-KATARQTJSA-N Thr-Ser-Lys Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCCN IQPWNQRRAJHOKV-KATARQTJSA-N 0.000 description 6
- YIKDYZDNRCNFQB-KKUMJFAQSA-N Tyr-His-Asn Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)O YIKDYZDNRCNFQB-KKUMJFAQSA-N 0.000 description 6
- XFEMMSGONWQACR-KJEVXHAQSA-N Tyr-Thr-Met Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCSC)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N)O XFEMMSGONWQACR-KJEVXHAQSA-N 0.000 description 6
- CELJCNRXKZPTCX-XPUUQOCRSA-N Val-Gly-Ala Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O CELJCNRXKZPTCX-XPUUQOCRSA-N 0.000 description 6
- XXROXFHCMVXETG-UWVGGRQHSA-N Val-Gly-Val Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O XXROXFHCMVXETG-UWVGGRQHSA-N 0.000 description 6
- 108010077245 asparaginyl-proline Proteins 0.000 description 6
- 108010047857 aspartylglycine Proteins 0.000 description 6
- 230000004071 biological effect Effects 0.000 description 6
- 238000010586 diagram Methods 0.000 description 6
- 108010026364 glycyl-glycyl-leucine Proteins 0.000 description 6
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 108010012581 phenylalanylglutamate Proteins 0.000 description 6
- 230000009257 reactivity Effects 0.000 description 6
- 108010048818 seryl-histidine Proteins 0.000 description 6
- 238000005809 transesterification reaction Methods 0.000 description 6
- NLOMBWNGESDVJU-GUBZILKMSA-N Ala-Met-Arg Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O NLOMBWNGESDVJU-GUBZILKMSA-N 0.000 description 5
- RZSLYUUFFVHFRQ-FXQIFTODSA-N Gln-Ala-Glu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O RZSLYUUFFVHFRQ-FXQIFTODSA-N 0.000 description 5
- RDPOETHPAQEGDP-ACZMJKKPSA-N Glu-Asp-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O RDPOETHPAQEGDP-ACZMJKKPSA-N 0.000 description 5
- FHQRLHFYVZAQHU-IUCAKERBSA-N Gly-Lys-Gln Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(O)=O FHQRLHFYVZAQHU-IUCAKERBSA-N 0.000 description 5
- QAMMIGULQSIRCD-IRXDYDNUSA-N Gly-Phe-Tyr Chemical compound C([C@H](NC(=O)C[NH3+])C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C([O-])=O)C1=CC=CC=C1 QAMMIGULQSIRCD-IRXDYDNUSA-N 0.000 description 5
- MDOBWSFNSNPENN-PMVVWTBXSA-N His-Thr-Gly Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O MDOBWSFNSNPENN-PMVVWTBXSA-N 0.000 description 5
- RQZFWBLDTBDEOF-RNJOBUHISA-N Ile-Val-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N RQZFWBLDTBDEOF-RNJOBUHISA-N 0.000 description 5
- JZBVBOKASHNXAD-NAKRPEOUSA-N Ile-Val-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(=O)O)N JZBVBOKASHNXAD-NAKRPEOUSA-N 0.000 description 5
- HVAUKHLDSDDROB-KKUMJFAQSA-N Lys-Lys-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O HVAUKHLDSDDROB-KKUMJFAQSA-N 0.000 description 5
- LECIJRIRMVOFMH-ULQDDVLXSA-N Lys-Pro-Phe Chemical compound NCCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 LECIJRIRMVOFMH-ULQDDVLXSA-N 0.000 description 5
- VHGIWFGJIHTASW-FXQIFTODSA-N Met-Ala-Asp Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O VHGIWFGJIHTASW-FXQIFTODSA-N 0.000 description 5
- SITLTJHOQZFJGG-UHFFFAOYSA-N N-L-alpha-glutamyl-L-valine Natural products CC(C)C(C(O)=O)NC(=O)C(N)CCC(O)=O SITLTJHOQZFJGG-UHFFFAOYSA-N 0.000 description 5
- 101100342977 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) leu-1 gene Proteins 0.000 description 5
- YYKZDTVQHTUKDW-RYUDHWBXSA-N Phe-Gly-Gln Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)NCC(=O)N[C@@H](CCC(=O)N)C(=O)O)N YYKZDTVQHTUKDW-RYUDHWBXSA-N 0.000 description 5
- VLMIUSLQONKLDV-HEIBUPTGSA-N Ser-Thr-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O VLMIUSLQONKLDV-HEIBUPTGSA-N 0.000 description 5
- NDZYTIMDOZMECO-SHGPDSBTSA-N Thr-Thr-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O NDZYTIMDOZMECO-SHGPDSBTSA-N 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 238000000354 decomposition reaction Methods 0.000 description 5
- 235000021588 free fatty acids Nutrition 0.000 description 5
- 108010089804 glycyl-threonine Proteins 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 239000002245 particle Substances 0.000 description 5
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 5
- 108010061238 threonyl-glycine Proteins 0.000 description 5
- PKOHVHWNGUHYRE-ZFWWWQNUSA-N (2s)-1-[2-[[(2s)-2-amino-3-(1h-indol-3-yl)propanoyl]amino]acetyl]pyrrolidine-2-carboxylic acid Chemical compound O=C([C@H](CC=1C2=CC=CC=C2NC=1)N)NCC(=O)N1CCC[C@H]1C(O)=O PKOHVHWNGUHYRE-ZFWWWQNUSA-N 0.000 description 4
- JBGSZRYCXBPWGX-BQBZGAKWSA-N Ala-Arg-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](N)C)CCCN=C(N)N JBGSZRYCXBPWGX-BQBZGAKWSA-N 0.000 description 4
- FXKNPWNXPQZLES-ZLUOBGJFSA-N Ala-Asn-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O FXKNPWNXPQZLES-ZLUOBGJFSA-N 0.000 description 4
- NIZKGBJVCMRDKO-KWQFWETISA-N Ala-Gly-Tyr Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 NIZKGBJVCMRDKO-KWQFWETISA-N 0.000 description 4
- NYDBKUNVSALYPX-NAKRPEOUSA-N Ala-Ile-Arg Chemical compound C[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CCCN=C(N)N NYDBKUNVSALYPX-NAKRPEOUSA-N 0.000 description 4
- AWZKCUCQJNTBAD-SRVKXCTJSA-N Ala-Leu-Lys Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCCN AWZKCUCQJNTBAD-SRVKXCTJSA-N 0.000 description 4
- VHAQSYHSDKERBS-XPUUQOCRSA-N Ala-Val-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O VHAQSYHSDKERBS-XPUUQOCRSA-N 0.000 description 4
- DCGLNNVKIZXQOJ-FXQIFTODSA-N Arg-Asn-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CCCN=C(N)N)N DCGLNNVKIZXQOJ-FXQIFTODSA-N 0.000 description 4
- KXEGPPNPXOKKHK-ZLUOBGJFSA-N Asn-Asp-Ala Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O KXEGPPNPXOKKHK-ZLUOBGJFSA-N 0.000 description 4
- PTSDPWIHOYMRGR-UGYAYLCHSA-N Asn-Ile-Asn Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(O)=O PTSDPWIHOYMRGR-UGYAYLCHSA-N 0.000 description 4
- DATSKXOXPUAOLK-KKUMJFAQSA-N Asn-Tyr-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(O)=O DATSKXOXPUAOLK-KKUMJFAQSA-N 0.000 description 4
- LDGUZSIPGSPBJP-XVYDVKMFSA-N Asp-His-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)[C@H](CC(=O)O)N LDGUZSIPGSPBJP-XVYDVKMFSA-N 0.000 description 4
- KLYPOCBLKMPBIQ-GHCJXIJMSA-N Asp-Ile-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CC(=O)O)N KLYPOCBLKMPBIQ-GHCJXIJMSA-N 0.000 description 4
- LIQNMKIBMPEOOP-IHRRRGAJSA-N Asp-Phe-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)[C@H](CC(=O)O)N LIQNMKIBMPEOOP-IHRRRGAJSA-N 0.000 description 4
- JSHWXQIZOCVWIA-ZKWXMUAHSA-N Asp-Ser-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O JSHWXQIZOCVWIA-ZKWXMUAHSA-N 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 4
- OZEQPCDLCDRCGY-SOUVJXGZSA-N Gln-Phe-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=CC=C2)NC(=O)[C@H](CCC(=O)N)N)C(=O)O OZEQPCDLCDRCGY-SOUVJXGZSA-N 0.000 description 4
- OCQUNKSFDYDXBG-QXEWZRGKSA-N Gly-Arg-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CCCN=C(N)N OCQUNKSFDYDXBG-QXEWZRGKSA-N 0.000 description 4
- ZQIMMEYPEXIYBB-IUCAKERBSA-N Gly-Glu-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)CN ZQIMMEYPEXIYBB-IUCAKERBSA-N 0.000 description 4
- OQQKUTVULYLCDG-ONGXEEELSA-N Gly-Lys-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CCCCN)NC(=O)CN)C(O)=O OQQKUTVULYLCDG-ONGXEEELSA-N 0.000 description 4
- IRJWAYCXIYUHQE-WHFBIAKZSA-N Gly-Ser-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)CN IRJWAYCXIYUHQE-WHFBIAKZSA-N 0.000 description 4
- LCRDMSSAKLTKBU-ZDLURKLDSA-N Gly-Ser-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)CN LCRDMSSAKLTKBU-ZDLURKLDSA-N 0.000 description 4
- 241000282414 Homo sapiens Species 0.000 description 4
- JHCVYQKVKOLAIU-NAKRPEOUSA-N Ile-Cys-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(=O)O)N JHCVYQKVKOLAIU-NAKRPEOUSA-N 0.000 description 4
- MQFGXJNSUJTXDT-QSFUFRPTSA-N Ile-Gly-Ile Chemical compound N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)O MQFGXJNSUJTXDT-QSFUFRPTSA-N 0.000 description 4
- YGDWPQCLFJNMOL-MNXVOIDGSA-N Ile-Leu-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N YGDWPQCLFJNMOL-MNXVOIDGSA-N 0.000 description 4
- YKZAMJXNJUWFIK-JBDRJPRFSA-N Ile-Ser-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(=O)O)N YKZAMJXNJUWFIK-JBDRJPRFSA-N 0.000 description 4
- KBDIBHQICWDGDL-PPCPHDFISA-N Ile-Thr-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)O)N KBDIBHQICWDGDL-PPCPHDFISA-N 0.000 description 4
- WSGXUIQTEZDVHJ-GARJFASQSA-N Leu-Ala-Pro Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@@H]1C(O)=O WSGXUIQTEZDVHJ-GARJFASQSA-N 0.000 description 4
- OXKYZSRZKBTVEY-ZPFDUUQYSA-N Leu-Asn-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O OXKYZSRZKBTVEY-ZPFDUUQYSA-N 0.000 description 4
- YOKVEHGYYQEQOP-QWRGUYRKSA-N Leu-Leu-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O YOKVEHGYYQEQOP-QWRGUYRKSA-N 0.000 description 4
- PTRKPHUGYULXPU-KKUMJFAQSA-N Leu-Phe-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O PTRKPHUGYULXPU-KKUMJFAQSA-N 0.000 description 4
- YWFZWQKWNDOWPA-XIRDDKMYSA-N Leu-Trp-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(N)=O)C(O)=O YWFZWQKWNDOWPA-XIRDDKMYSA-N 0.000 description 4
- AAKRWBIIGKPOKQ-ONGXEEELSA-N Leu-Val-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O AAKRWBIIGKPOKQ-ONGXEEELSA-N 0.000 description 4
- WQWZXKWOEVSGQM-DCAQKATOSA-N Lys-Ala-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN WQWZXKWOEVSGQM-DCAQKATOSA-N 0.000 description 4
- MQASRXPTQJJNFM-JYJNAYRXSA-N Met-Pro-Phe Chemical compound CSCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 MQASRXPTQJJNFM-JYJNAYRXSA-N 0.000 description 4
- 235000019482 Palm oil Nutrition 0.000 description 4
- DLZBBDSPTJBOOD-BPNCWPANSA-N Pro-Tyr-Ala Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C)C(O)=O DLZBBDSPTJBOOD-BPNCWPANSA-N 0.000 description 4
- XWCYBVBLJRWOFR-WDSKDSINSA-N Ser-Gln-Gly Chemical compound OC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O XWCYBVBLJRWOFR-WDSKDSINSA-N 0.000 description 4
- CICQXRWZNVXFCU-SRVKXCTJSA-N Ser-His-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(C)C)C(O)=O CICQXRWZNVXFCU-SRVKXCTJSA-N 0.000 description 4
- WUXCHQZLUHBSDJ-LKXGYXEUSA-N Ser-Thr-Asp Chemical compound OC[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CC(O)=O)C(O)=O WUXCHQZLUHBSDJ-LKXGYXEUSA-N 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- VXMHQKHDKCATDV-VEVYYDQMSA-N Thr-Asp-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O VXMHQKHDKCATDV-VEVYYDQMSA-N 0.000 description 4
- UYTYTDMCDBPDSC-URLPEUOOSA-N Thr-Ile-Phe Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H]([C@@H](C)O)N UYTYTDMCDBPDSC-URLPEUOOSA-N 0.000 description 4
- VEIKMWOMUYMMMK-FCLVOEFKSA-N Thr-Phe-Phe Chemical compound C([C@H](NC(=O)[C@@H](N)[C@H](O)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 VEIKMWOMUYMMMK-FCLVOEFKSA-N 0.000 description 4
- QUILOGWWLXMSAT-IHRRRGAJSA-N Tyr-Gln-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O QUILOGWWLXMSAT-IHRRRGAJSA-N 0.000 description 4
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 4
- JZWZACGUZVCQPS-RNJOBUHISA-N Val-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](C(C)C)N JZWZACGUZVCQPS-RNJOBUHISA-N 0.000 description 4
- LTTQCQRTSHJPPL-ZKWXMUAHSA-N Val-Ser-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(=O)O)C(=O)O)N LTTQCQRTSHJPPL-ZKWXMUAHSA-N 0.000 description 4
- BGTDGENDNWGMDQ-KJEVXHAQSA-N Val-Tyr-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](C(C)C)N)O BGTDGENDNWGMDQ-KJEVXHAQSA-N 0.000 description 4
- 230000002255 enzymatic effect Effects 0.000 description 4
- 108010087823 glycyltyrosine Proteins 0.000 description 4
- 230000002209 hydrophobic effect Effects 0.000 description 4
- 125000001165 hydrophobic group Chemical group 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 239000002540 palm oil Substances 0.000 description 4
- 108010053725 prolylvaline Proteins 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- 238000004809 thin layer chromatography Methods 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- GORKKVHIBWAQHM-GCJQMDKQSA-N Ala-Asn-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GORKKVHIBWAQHM-GCJQMDKQSA-N 0.000 description 3
- MKJBPDLENBUHQU-CIUDSAMLSA-N Asn-Ser-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O MKJBPDLENBUHQU-CIUDSAMLSA-N 0.000 description 3
- BUVNWKQBMZLCDW-UGYAYLCHSA-N Asp-Asn-Ile Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O BUVNWKQBMZLCDW-UGYAYLCHSA-N 0.000 description 3
- KYQNAIMCTRZLNP-QSFUFRPTSA-N Asp-Ile-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(O)=O KYQNAIMCTRZLNP-QSFUFRPTSA-N 0.000 description 3
- CLUMZOKVGUWUFD-CIUDSAMLSA-N Asp-Leu-Asn Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O CLUMZOKVGUWUFD-CIUDSAMLSA-N 0.000 description 3
- GIKOVDMXBAFXDF-NHCYSSNCSA-N Asp-Val-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O GIKOVDMXBAFXDF-NHCYSSNCSA-N 0.000 description 3
- 241000699800 Cricetinae Species 0.000 description 3
- 108010074860 Factor Xa Proteins 0.000 description 3
- PAWIVEIWWYGBAM-YUMQZZPRSA-N Gly-Leu-Ala Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O PAWIVEIWWYGBAM-YUMQZZPRSA-N 0.000 description 3
- RGPWUJOMKFYFSR-QWRGUYRKSA-N His-Gly-Leu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)NCC(=O)N[C@@H](CC(C)C)C(O)=O RGPWUJOMKFYFSR-QWRGUYRKSA-N 0.000 description 3
- JUWJEAPUNARGCF-DCAQKATOSA-N Leu-Arg-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(O)=O JUWJEAPUNARGCF-DCAQKATOSA-N 0.000 description 3
- PPGBXYKMUMHFBF-KATARQTJSA-N Leu-Ser-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PPGBXYKMUMHFBF-KATARQTJSA-N 0.000 description 3
- VHTIZYYHIUHMCA-JYJNAYRXSA-N Leu-Tyr-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(O)=O VHTIZYYHIUHMCA-JYJNAYRXSA-N 0.000 description 3
- WSXTWLJHTLRFLW-SRVKXCTJSA-N Lys-Ala-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O WSXTWLJHTLRFLW-SRVKXCTJSA-N 0.000 description 3
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 3
- WYBVBIHNJWOLCJ-UHFFFAOYSA-N N-L-arginyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCCN=C(N)N WYBVBIHNJWOLCJ-UHFFFAOYSA-N 0.000 description 3
- CGOMLCQJEMWMCE-STQMWFEESA-N Phe-Arg-Gly Chemical compound NC(N)=NCCC[C@@H](C(=O)NCC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 CGOMLCQJEMWMCE-STQMWFEESA-N 0.000 description 3
- VXCHGLYSIOOZIS-GUBZILKMSA-N Pro-Ala-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1 VXCHGLYSIOOZIS-GUBZILKMSA-N 0.000 description 3
- BODDREDDDRZUCF-QTKMDUPCSA-N Pro-His-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)[C@@H]2CCCN2)O BODDREDDDRZUCF-QTKMDUPCSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- VDUJEEQMRQCLHB-YTQUADARSA-N Trp-Lys-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCCN)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)N)C(=O)O VDUJEEQMRQCLHB-YTQUADARSA-N 0.000 description 3
- ZZDFLJFVSNQINX-HWHUXHBOSA-N Trp-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)N)O ZZDFLJFVSNQINX-HWHUXHBOSA-N 0.000 description 3
- UUZYQOUJTORBQO-ZVZYQTTQSA-N Trp-Val-Gln Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O)=CNC2=C1 UUZYQOUJTORBQO-ZVZYQTTQSA-N 0.000 description 3
- JLFKWDAZBRYCGX-ZKWXMUAHSA-N Val-Asn-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CO)C(=O)O)N JLFKWDAZBRYCGX-ZKWXMUAHSA-N 0.000 description 3
- JTWIMNMUYLQNPI-WPRPVWTQSA-N Val-Gly-Arg Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCNC(N)=N JTWIMNMUYLQNPI-WPRPVWTQSA-N 0.000 description 3
- UMPVMAYCLYMYGA-ONGXEEELSA-N Val-Leu-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O UMPVMAYCLYMYGA-ONGXEEELSA-N 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 108010044940 alanylglutamine Proteins 0.000 description 3
- BTANRVKWQNVYAZ-UHFFFAOYSA-N butan-2-ol Chemical compound CCC(C)O BTANRVKWQNVYAZ-UHFFFAOYSA-N 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- 239000003054 catalyst Substances 0.000 description 3
- 230000003197 catalytic effect Effects 0.000 description 3
- 238000006731 degradation reaction Methods 0.000 description 3
- 235000014113 dietary fatty acids Nutrition 0.000 description 3
- 230000001747 exhibiting effect Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 229930195729 fatty acid Natural products 0.000 description 3
- 239000000194 fatty acid Substances 0.000 description 3
- 150000004665 fatty acids Chemical class 0.000 description 3
- 238000004817 gas chromatography Methods 0.000 description 3
- 235000011187 glycerol Nutrition 0.000 description 3
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 3
- 210000003000 inclusion body Anatomy 0.000 description 3
- 210000003292 kidney cell Anatomy 0.000 description 3
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 230000036962 time dependent Effects 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- RLMISHABBKUNFO-WHFBIAKZSA-N Ala-Ala-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O RLMISHABBKUNFO-WHFBIAKZSA-N 0.000 description 2
- MKZCBYZBCINNJN-DLOVCJGASA-N Ala-Asp-Phe Chemical compound C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 MKZCBYZBCINNJN-DLOVCJGASA-N 0.000 description 2
- BTYTYHBSJKQBQA-GCJQMDKQSA-N Ala-Asp-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](C)N)O BTYTYHBSJKQBQA-GCJQMDKQSA-N 0.000 description 2
- KUDREHRZRIVKHS-UWJYBYFXSA-N Ala-Asp-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O KUDREHRZRIVKHS-UWJYBYFXSA-N 0.000 description 2
- ZVFVBBGVOILKPO-WHFBIAKZSA-N Ala-Gly-Ala Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O ZVFVBBGVOILKPO-WHFBIAKZSA-N 0.000 description 2
- BLIMFWGRQKRCGT-YUMQZZPRSA-N Ala-Gly-Lys Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCCN BLIMFWGRQKRCGT-YUMQZZPRSA-N 0.000 description 2
- MQIGTEQXYCRLGK-BQBZGAKWSA-N Ala-Gly-Pro Chemical compound C[C@H](N)C(=O)NCC(=O)N1CCC[C@H]1C(O)=O MQIGTEQXYCRLGK-BQBZGAKWSA-N 0.000 description 2
- GRPHQEMIFDPKOE-HGNGGELXSA-N Ala-His-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(O)=O)C(O)=O GRPHQEMIFDPKOE-HGNGGELXSA-N 0.000 description 2
- PNALXAODQKTNLV-JBDRJPRFSA-N Ala-Ile-Ala Chemical compound C[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O PNALXAODQKTNLV-JBDRJPRFSA-N 0.000 description 2
- LBYMZCVBOKYZNS-CIUDSAMLSA-N Ala-Leu-Asp Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O LBYMZCVBOKYZNS-CIUDSAMLSA-N 0.000 description 2
- SOBIAADAMRHGKH-CIUDSAMLSA-N Ala-Leu-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O SOBIAADAMRHGKH-CIUDSAMLSA-N 0.000 description 2
- AJBVYEYZVYPFCF-CIUDSAMLSA-N Ala-Lys-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O AJBVYEYZVYPFCF-CIUDSAMLSA-N 0.000 description 2
- WQKAQKZRDIZYNV-VZFHVOOUSA-N Ala-Ser-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O WQKAQKZRDIZYNV-VZFHVOOUSA-N 0.000 description 2
- OEVCHROQUIVQFZ-YTLHQDLWSA-N Ala-Thr-Ala Chemical compound C[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](C)C(O)=O OEVCHROQUIVQFZ-YTLHQDLWSA-N 0.000 description 2
- AENHOIXXHKNIQL-AUTRQRHGSA-N Ala-Tyr-Ala Chemical compound [O-]C(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@@H]([NH3+])C)CC1=CC=C(O)C=C1 AENHOIXXHKNIQL-AUTRQRHGSA-N 0.000 description 2
- CLOMBHBBUKAUBP-LSJOCFKGSA-N Ala-Val-His Chemical compound C[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N CLOMBHBBUKAUBP-LSJOCFKGSA-N 0.000 description 2
- PQWTZSNVWSOFFK-FXQIFTODSA-N Arg-Asp-Asn Chemical compound C(C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)CN=C(N)N PQWTZSNVWSOFFK-FXQIFTODSA-N 0.000 description 2
- PHHRSPBBQUFULD-UWVGGRQHSA-N Arg-Gly-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CCCN=C(N)N)N PHHRSPBBQUFULD-UWVGGRQHSA-N 0.000 description 2
- RZVVKNIACROXRM-ZLUOBGJFSA-N Asn-Ala-Asp Chemical compound C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CC(=O)N)N RZVVKNIACROXRM-ZLUOBGJFSA-N 0.000 description 2
- XQQVCUIBGYFKDC-OLHMAJIHSA-N Asn-Asp-Thr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XQQVCUIBGYFKDC-OLHMAJIHSA-N 0.000 description 2
- JQSWHKKUZMTOIH-QWRGUYRKSA-N Asn-Gly-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CC(=O)N)N JQSWHKKUZMTOIH-QWRGUYRKSA-N 0.000 description 2
- SPCONPVIDFMDJI-QSFUFRPTSA-N Asn-Ile-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(O)=O SPCONPVIDFMDJI-QSFUFRPTSA-N 0.000 description 2
- BXUHCIXDSWRSBS-CIUDSAMLSA-N Asn-Leu-Asp Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O BXUHCIXDSWRSBS-CIUDSAMLSA-N 0.000 description 2
- WIDVAWAQBRAKTI-YUMQZZPRSA-N Asn-Leu-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O WIDVAWAQBRAKTI-YUMQZZPRSA-N 0.000 description 2
- AYOAHKWVQLNPDM-HJGDQZAQSA-N Asn-Lys-Thr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O AYOAHKWVQLNPDM-HJGDQZAQSA-N 0.000 description 2
- PIABYSIYPGLLDQ-XVSYOHENSA-N Asn-Thr-Phe Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O PIABYSIYPGLLDQ-XVSYOHENSA-N 0.000 description 2
- KGAJCJXBEWLQDZ-UBHSHLNASA-N Asp-Asp-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC(=O)O)N KGAJCJXBEWLQDZ-UBHSHLNASA-N 0.000 description 2
- SMZCLQGDQMGESY-ACZMJKKPSA-N Asp-Gln-Cys Chemical compound C(CC(=O)N)[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC(=O)O)N SMZCLQGDQMGESY-ACZMJKKPSA-N 0.000 description 2
- RQYMKRMRZWJGHC-BQBZGAKWSA-N Asp-Gly-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CC(=O)O)N RQYMKRMRZWJGHC-BQBZGAKWSA-N 0.000 description 2
- SWTQDYFZVOJVLL-KKUMJFAQSA-N Asp-His-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CC2=CN=CN2)NC(=O)[C@H](CC(=O)O)N)O SWTQDYFZVOJVLL-KKUMJFAQSA-N 0.000 description 2
- SCQIQCWLOMOEFP-DCAQKATOSA-N Asp-Leu-Arg Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O SCQIQCWLOMOEFP-DCAQKATOSA-N 0.000 description 2
- MYLZFUMPZCPJCJ-NHCYSSNCSA-N Asp-Lys-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O MYLZFUMPZCPJCJ-NHCYSSNCSA-N 0.000 description 2
- DINOVZWPTMGSRF-QXEWZRGKSA-N Asp-Pro-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O DINOVZWPTMGSRF-QXEWZRGKSA-N 0.000 description 2
- KBJVTFWQWXCYCQ-IUKAMOBKSA-N Asp-Thr-Ile Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O KBJVTFWQWXCYCQ-IUKAMOBKSA-N 0.000 description 2
- 108091026890 Coding region Proteins 0.000 description 2
- 108020004705 Codon Proteins 0.000 description 2
- IZUNQDRIAOLWCN-YUMQZZPRSA-N Cys-Leu-Gly Chemical compound CC(C)C[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CS)N IZUNQDRIAOLWCN-YUMQZZPRSA-N 0.000 description 2
- 229920002307 Dextran Polymers 0.000 description 2
- WMOMPXKOKASNBK-PEFMBERDSA-N Gln-Asn-Ile Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WMOMPXKOKASNBK-PEFMBERDSA-N 0.000 description 2
- BTSPOOHJBYJRKO-CIUDSAMLSA-N Gln-Asp-Arg Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O BTSPOOHJBYJRKO-CIUDSAMLSA-N 0.000 description 2
- ORYMMTRPKVTGSJ-XVKPBYJWSA-N Gln-Gly-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCC(N)=O ORYMMTRPKVTGSJ-XVKPBYJWSA-N 0.000 description 2
- ATRHMOJQJWPVBQ-DRZSPHRISA-N Glu-Ala-Phe Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O ATRHMOJQJWPVBQ-DRZSPHRISA-N 0.000 description 2
- MXOODARRORARSU-ACZMJKKPSA-N Glu-Ala-Ser Chemical compound C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCC(=O)O)N MXOODARRORARSU-ACZMJKKPSA-N 0.000 description 2
- GYCPQVFKCPPRQB-GUBZILKMSA-N Glu-Gln-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCC(=O)O)N GYCPQVFKCPPRQB-GUBZILKMSA-N 0.000 description 2
- OGNJZUXUTPQVBR-BQBZGAKWSA-N Glu-Gly-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O OGNJZUXUTPQVBR-BQBZGAKWSA-N 0.000 description 2
- VGBSZQSKQRMLHD-MNXVOIDGSA-N Glu-Leu-Ile Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O VGBSZQSKQRMLHD-MNXVOIDGSA-N 0.000 description 2
- PAZQYODKOZHXGA-SRVKXCTJSA-N Glu-Pro-His Chemical compound N[C@@H](CCC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O PAZQYODKOZHXGA-SRVKXCTJSA-N 0.000 description 2
- UUTGYDAKPISJAO-JYJNAYRXSA-N Glu-Tyr-Leu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CC(C)C)C(O)=O)CC1=CC=C(O)C=C1 UUTGYDAKPISJAO-JYJNAYRXSA-N 0.000 description 2
- SOYWRINXUSUWEQ-DLOVCJGASA-N Glu-Val-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCC(O)=O SOYWRINXUSUWEQ-DLOVCJGASA-N 0.000 description 2
- FMNHBTKMRFVGRO-FOHZUACHSA-N Gly-Asn-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)CN FMNHBTKMRFVGRO-FOHZUACHSA-N 0.000 description 2
- FZQLXNIMCPJVJE-YUMQZZPRSA-N Gly-Asp-Leu Chemical compound [H]NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O FZQLXNIMCPJVJE-YUMQZZPRSA-N 0.000 description 2
- HPAIKDPJURGQLN-KBPBESRZSA-N Gly-His-Phe Chemical compound C([C@H](NC(=O)CN)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CNC=N1 HPAIKDPJURGQLN-KBPBESRZSA-N 0.000 description 2
- KGVHCTWYMPWEGN-FSPLSTOPSA-N Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)CN KGVHCTWYMPWEGN-FSPLSTOPSA-N 0.000 description 2
- TWTPDFFBLQEBOE-IUCAKERBSA-N Gly-Leu-Gln Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O TWTPDFFBLQEBOE-IUCAKERBSA-N 0.000 description 2
- UHPAZODVFFYEEL-QWRGUYRKSA-N Gly-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)CN UHPAZODVFFYEEL-QWRGUYRKSA-N 0.000 description 2
- VEPBEGNDJYANCF-QWRGUYRKSA-N Gly-Lys-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CCCCN VEPBEGNDJYANCF-QWRGUYRKSA-N 0.000 description 2
- FXLVSYVJDPCIHH-STQMWFEESA-N Gly-Phe-Arg Chemical compound [H]NCC(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O FXLVSYVJDPCIHH-STQMWFEESA-N 0.000 description 2
- OHUKZZYSJBKFRR-WHFBIAKZSA-N Gly-Ser-Asp Chemical compound [H]NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O OHUKZZYSJBKFRR-WHFBIAKZSA-N 0.000 description 2
- FKESCSGWBPUTPN-FOHZUACHSA-N Gly-Thr-Asn Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(O)=O FKESCSGWBPUTPN-FOHZUACHSA-N 0.000 description 2
- UMBDRSMLCUYIRI-DVJZZOLTSA-N Gly-Trp-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)CN)O UMBDRSMLCUYIRI-DVJZZOLTSA-N 0.000 description 2
- UVTSZKIATYSKIR-RYUDHWBXSA-N Gly-Tyr-Glu Chemical compound [H]NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O UVTSZKIATYSKIR-RYUDHWBXSA-N 0.000 description 2
- HQSKKSLNLSTONK-JTQLQIEISA-N Gly-Tyr-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)CN)CC1=CC=C(O)C=C1 HQSKKSLNLSTONK-JTQLQIEISA-N 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 2
- BDHUXUFYNUOUIT-SRVKXCTJSA-N His-Asp-Lys Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N BDHUXUFYNUOUIT-SRVKXCTJSA-N 0.000 description 2
- BSVLMPMIXPQNKC-KBPBESRZSA-N His-Phe-Gly Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)NCC(O)=O BSVLMPMIXPQNKC-KBPBESRZSA-N 0.000 description 2
- LKACSKJPTFSBHR-MNXVOIDGSA-N Ile-Gln-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CCCCN)C(=O)O)N LKACSKJPTFSBHR-MNXVOIDGSA-N 0.000 description 2
- MTFVYKQRLXYAQN-LAEOZQHASA-N Ile-Glu-Gly Chemical compound [H]N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O MTFVYKQRLXYAQN-LAEOZQHASA-N 0.000 description 2
- KFVUBLZRFSVDGO-BYULHYEWSA-N Ile-Gly-Asp Chemical compound CC[C@H](C)[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC(O)=O KFVUBLZRFSVDGO-BYULHYEWSA-N 0.000 description 2
- YNMQUIVKEFRCPH-QSFUFRPTSA-N Ile-Ile-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)O)N YNMQUIVKEFRCPH-QSFUFRPTSA-N 0.000 description 2
- GAZGFPOZOLEYAJ-YTFOTSKYSA-N Ile-Leu-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N GAZGFPOZOLEYAJ-YTFOTSKYSA-N 0.000 description 2
- CIJLNXXMDUOFPH-HJWJTTGWSA-N Ile-Pro-Phe Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 CIJLNXXMDUOFPH-HJWJTTGWSA-N 0.000 description 2
- YJRSIJZUIUANHO-NAKRPEOUSA-N Ile-Val-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(=O)O)N YJRSIJZUIUANHO-NAKRPEOUSA-N 0.000 description 2
- 108010065920 Insulin Lispro Proteins 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 2
- RCFDOSNHHZGBOY-UHFFFAOYSA-N L-isoleucyl-L-alanine Natural products CCC(C)C(N)C(=O)NC(C)C(O)=O RCFDOSNHHZGBOY-UHFFFAOYSA-N 0.000 description 2
- KFKWRHQBZQICHA-STQMWFEESA-N L-leucyl-L-phenylalanine Natural products CC(C)C[C@H](N)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 KFKWRHQBZQICHA-STQMWFEESA-N 0.000 description 2
- LZDNBBYBDGBADK-UHFFFAOYSA-N L-valyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)C(C)C)C(O)=O)=CNC2=C1 LZDNBBYBDGBADK-UHFFFAOYSA-N 0.000 description 2
- NTRAGDHVSGKUSF-AVGNSLFASA-N Leu-Arg-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O NTRAGDHVSGKUSF-AVGNSLFASA-N 0.000 description 2
- KKXDHFKZWKLYGB-GUBZILKMSA-N Leu-Asn-Glu Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N KKXDHFKZWKLYGB-GUBZILKMSA-N 0.000 description 2
- KTFHTMHHKXUYPW-ZPFDUUQYSA-N Leu-Asp-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O KTFHTMHHKXUYPW-ZPFDUUQYSA-N 0.000 description 2
- NEEOBPIXKWSBRF-IUCAKERBSA-N Leu-Glu-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O NEEOBPIXKWSBRF-IUCAKERBSA-N 0.000 description 2
- WQWSMEOYXJTFRU-GUBZILKMSA-N Leu-Glu-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O WQWSMEOYXJTFRU-GUBZILKMSA-N 0.000 description 2
- APFJUBGRZGMQFF-QWRGUYRKSA-N Leu-Gly-Lys Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCCN APFJUBGRZGMQFF-QWRGUYRKSA-N 0.000 description 2
- POZULHZYLPGXMR-ONGXEEELSA-N Leu-Gly-Val Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O POZULHZYLPGXMR-ONGXEEELSA-N 0.000 description 2
- AUBMZAMQCOYSIC-MNXVOIDGSA-N Leu-Ile-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(N)=O)C(O)=O AUBMZAMQCOYSIC-MNXVOIDGSA-N 0.000 description 2
- QNBVTHNJGCOVFA-AVGNSLFASA-N Leu-Leu-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCC(O)=O QNBVTHNJGCOVFA-AVGNSLFASA-N 0.000 description 2
- LVTJJOJKDCVZGP-QWRGUYRKSA-N Leu-Lys-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O LVTJJOJKDCVZGP-QWRGUYRKSA-N 0.000 description 2
- FKQPWMZLIIATBA-AJNGGQMLSA-N Leu-Lys-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O FKQPWMZLIIATBA-AJNGGQMLSA-N 0.000 description 2
- RTIRBWJPYJYTLO-MELADBBJSA-N Leu-Lys-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N1CCC[C@@H]1C(=O)O)N RTIRBWJPYJYTLO-MELADBBJSA-N 0.000 description 2
- FLNPJLDPGMLWAU-UWVGGRQHSA-N Leu-Met-Gly Chemical compound OC(=O)CNC(=O)[C@H](CCSC)NC(=O)[C@@H](N)CC(C)C FLNPJLDPGMLWAU-UWVGGRQHSA-N 0.000 description 2
- DDVHDMSBLRAKNV-IHRRRGAJSA-N Leu-Met-Leu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(C)C)C(O)=O DDVHDMSBLRAKNV-IHRRRGAJSA-N 0.000 description 2
- BIZNDKMFQHDOIE-KKUMJFAQSA-N Leu-Phe-Asn Chemical compound CC(C)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CC(N)=O)C(O)=O)CC1=CC=CC=C1 BIZNDKMFQHDOIE-KKUMJFAQSA-N 0.000 description 2
- XXXXOVFBXRERQL-ULQDDVLXSA-N Leu-Pro-Phe Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 XXXXOVFBXRERQL-ULQDDVLXSA-N 0.000 description 2
- BRTVHXHCUSXYRI-CIUDSAMLSA-N Leu-Ser-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O BRTVHXHCUSXYRI-CIUDSAMLSA-N 0.000 description 2
- PNPYKQFJGRFYJE-GUBZILKMSA-N Lys-Ala-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O PNPYKQFJGRFYJE-GUBZILKMSA-N 0.000 description 2
- IRNSXVOWSXSULE-DCAQKATOSA-N Lys-Ala-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN IRNSXVOWSXSULE-DCAQKATOSA-N 0.000 description 2
- DGAAQRAUOFHBFJ-CIUDSAMLSA-N Lys-Asn-Ala Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(O)=O DGAAQRAUOFHBFJ-CIUDSAMLSA-N 0.000 description 2
- HKCCVDWHHTVVPN-CIUDSAMLSA-N Lys-Asp-Ala Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O HKCCVDWHHTVVPN-CIUDSAMLSA-N 0.000 description 2
- WGCKDDHUFPQSMZ-ZPFDUUQYSA-N Lys-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CCCCN WGCKDDHUFPQSMZ-ZPFDUUQYSA-N 0.000 description 2
- VSRXPEHZMHSFKU-IUCAKERBSA-N Lys-Gln-Gly Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O VSRXPEHZMHSFKU-IUCAKERBSA-N 0.000 description 2
- YVMQJGWLHRWMDF-MNXVOIDGSA-N Lys-Gln-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCCCN)N YVMQJGWLHRWMDF-MNXVOIDGSA-N 0.000 description 2
- ITWQLSZTLBKWJM-YUMQZZPRSA-N Lys-Gly-Ala Chemical compound OC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H](N)CCCCN ITWQLSZTLBKWJM-YUMQZZPRSA-N 0.000 description 2
- ISHNZELVUVPCHY-ZETCQYMHSA-N Lys-Gly-Gly Chemical compound NCCCC[C@H](N)C(=O)NCC(=O)NCC(O)=O ISHNZELVUVPCHY-ZETCQYMHSA-N 0.000 description 2
- IVFUVMSKSFSFBT-NHCYSSNCSA-N Lys-Ile-Gly Chemical compound OC(=O)CNC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CCCCN IVFUVMSKSFSFBT-NHCYSSNCSA-N 0.000 description 2
- PRSBSVAVOQOAMI-BJDJZHNGSA-N Lys-Ile-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CCCCN PRSBSVAVOQOAMI-BJDJZHNGSA-N 0.000 description 2
- WRODMZBHNNPRLN-SRVKXCTJSA-N Lys-Leu-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O WRODMZBHNNPRLN-SRVKXCTJSA-N 0.000 description 2
- RIJCHEVHFWMDKD-SRVKXCTJSA-N Lys-Lys-Asn Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O RIJCHEVHFWMDKD-SRVKXCTJSA-N 0.000 description 2
- BEGQVWUZFXLNHZ-IHPCNDPISA-N Lys-Lys-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CCCCN)C(O)=O)=CNC2=C1 BEGQVWUZFXLNHZ-IHPCNDPISA-N 0.000 description 2
- LUTDBHBIHHREDC-IHRRRGAJSA-N Lys-Pro-Lys Chemical compound NCCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(O)=O LUTDBHBIHHREDC-IHRRRGAJSA-N 0.000 description 2
- JOSAKOKSPXROGQ-BJDJZHNGSA-N Lys-Ser-Ile Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JOSAKOKSPXROGQ-BJDJZHNGSA-N 0.000 description 2
- RPWTZTBIFGENIA-VOAKCMCISA-N Lys-Thr-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O RPWTZTBIFGENIA-VOAKCMCISA-N 0.000 description 2
- CNXOBMMOYZPPGS-NUTKFTJISA-N Lys-Trp-Ala Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](C)C(O)=O CNXOBMMOYZPPGS-NUTKFTJISA-N 0.000 description 2
- WDTLNWHPIPCMMP-AVGNSLFASA-N Met-Arg-Leu Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O WDTLNWHPIPCMMP-AVGNSLFASA-N 0.000 description 2
- WPTHAGXMYDRPFD-SRVKXCTJSA-N Met-Lys-Glu Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O WPTHAGXMYDRPFD-SRVKXCTJSA-N 0.000 description 2
- HSJIGJRZYUADSS-IHRRRGAJSA-N Met-Lys-Leu Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O HSJIGJRZYUADSS-IHRRRGAJSA-N 0.000 description 2
- QTMIXEQWGNIPBL-JYJNAYRXSA-N Met-Met-Tyr Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N QTMIXEQWGNIPBL-JYJNAYRXSA-N 0.000 description 2
- MUDYEFAKNSTFAI-JYJNAYRXSA-N Met-Tyr-Val Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C(C)C)C(O)=O MUDYEFAKNSTFAI-JYJNAYRXSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- WUGMRIBZSVSJNP-UHFFFAOYSA-N N-L-alanyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)C)C(O)=O)=CNC2=C1 WUGMRIBZSVSJNP-UHFFFAOYSA-N 0.000 description 2
- AFCARXCZXQIEQB-UHFFFAOYSA-N N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CCNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 AFCARXCZXQIEQB-UHFFFAOYSA-N 0.000 description 2
- AJHCSUXXECOXOY-UHFFFAOYSA-N N-glycyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)CN)C(O)=O)=CNC2=C1 AJHCSUXXECOXOY-UHFFFAOYSA-N 0.000 description 2
- 108010066427 N-valyltryptophan Proteins 0.000 description 2
- 101100068676 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) gln-1 gene Proteins 0.000 description 2
- 108700026244 Open Reading Frames Proteins 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- DPUOLKQSMYLRDR-UBHSHLNASA-N Phe-Arg-Ala Chemical compound NC(N)=NCCC[C@@H](C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 DPUOLKQSMYLRDR-UBHSHLNASA-N 0.000 description 2
- ZENDEDYRYVHBEG-SRVKXCTJSA-N Phe-Asp-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 ZENDEDYRYVHBEG-SRVKXCTJSA-N 0.000 description 2
- UNLYPPYNDXHGDG-IHRRRGAJSA-N Phe-Gln-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 UNLYPPYNDXHGDG-IHRRRGAJSA-N 0.000 description 2
- MPFGIYLYWUCSJG-AVGNSLFASA-N Phe-Glu-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 MPFGIYLYWUCSJG-AVGNSLFASA-N 0.000 description 2
- JJHVFCUWLSKADD-ONGXEEELSA-N Phe-Gly-Ala Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H](C)C(O)=O JJHVFCUWLSKADD-ONGXEEELSA-N 0.000 description 2
- GXDPQJUBLBZKDY-IAVJCBSLSA-N Phe-Ile-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O GXDPQJUBLBZKDY-IAVJCBSLSA-N 0.000 description 2
- KBVJZCVLQWCJQN-KKUMJFAQSA-N Phe-Leu-Asn Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O KBVJZCVLQWCJQN-KKUMJFAQSA-N 0.000 description 2
- ZLAKUZDMKVKFAI-JYJNAYRXSA-N Phe-Pro-Val Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O ZLAKUZDMKVKFAI-JYJNAYRXSA-N 0.000 description 2
- XNMYNGDKJNOKHH-BZSNNMDCSA-N Phe-Ser-Tyr Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O XNMYNGDKJNOKHH-BZSNNMDCSA-N 0.000 description 2
- 206010035226 Plasma cell myeloma Diseases 0.000 description 2
- HFZNNDWPHBRNPV-KZVJFYERSA-N Pro-Ala-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O HFZNNDWPHBRNPV-KZVJFYERSA-N 0.000 description 2
- IHCXPSYCHXFXKT-DCAQKATOSA-N Pro-Arg-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O IHCXPSYCHXFXKT-DCAQKATOSA-N 0.000 description 2
- NMELOOXSGDRBRU-YUMQZZPRSA-N Pro-Glu-Gly Chemical compound OC(=O)CNC(=O)[C@H](CCC(=O)O)NC(=O)[C@@H]1CCCN1 NMELOOXSGDRBRU-YUMQZZPRSA-N 0.000 description 2
- VYWNORHENYEQDW-YUMQZZPRSA-N Pro-Gly-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H]1CCCN1 VYWNORHENYEQDW-YUMQZZPRSA-N 0.000 description 2
- VZKBJNBZMZHKRC-XUXIUFHCSA-N Pro-Ile-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O VZKBJNBZMZHKRC-XUXIUFHCSA-N 0.000 description 2
- UREQLMJCKFLLHM-NAKRPEOUSA-N Pro-Ile-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O UREQLMJCKFLLHM-NAKRPEOUSA-N 0.000 description 2
- OFGUOWQVEGTVNU-DCAQKATOSA-N Pro-Lys-Ala Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O OFGUOWQVEGTVNU-DCAQKATOSA-N 0.000 description 2
- POQFNPILEQEODH-FXQIFTODSA-N Pro-Ser-Ala Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O POQFNPILEQEODH-FXQIFTODSA-N 0.000 description 2
- HRIXMVRZRGFKNQ-HJGDQZAQSA-N Pro-Thr-Gln Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(O)=O HRIXMVRZRGFKNQ-HJGDQZAQSA-N 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 2
- BTKUIVBNGBFTTP-WHFBIAKZSA-N Ser-Ala-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)NCC(O)=O BTKUIVBNGBFTTP-WHFBIAKZSA-N 0.000 description 2
- HRNQLKCLPVKZNE-CIUDSAMLSA-N Ser-Ala-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O HRNQLKCLPVKZNE-CIUDSAMLSA-N 0.000 description 2
- IYCBDVBJWDXQRR-FXQIFTODSA-N Ser-Ala-Met Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CCSC)C(O)=O IYCBDVBJWDXQRR-FXQIFTODSA-N 0.000 description 2
- PZZJMBYSYAKYPK-UWJYBYFXSA-N Ser-Ala-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O PZZJMBYSYAKYPK-UWJYBYFXSA-N 0.000 description 2
- NJSPTZXVPZDRCU-UBHSHLNASA-N Ser-Asp-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CO)N NJSPTZXVPZDRCU-UBHSHLNASA-N 0.000 description 2
- QKQDTEYDEIJPNK-GUBZILKMSA-N Ser-Glu-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CO QKQDTEYDEIJPNK-GUBZILKMSA-N 0.000 description 2
- UGHCUDLCCVVIJR-VGDYDELISA-N Ser-His-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)[C@H](CO)N UGHCUDLCCVVIJR-VGDYDELISA-N 0.000 description 2
- SFTZTYBXIXLRGQ-JBDRJPRFSA-N Ser-Ile-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O SFTZTYBXIXLRGQ-JBDRJPRFSA-N 0.000 description 2
- HEUVHBXOVZONPU-BJDJZHNGSA-N Ser-Leu-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O HEUVHBXOVZONPU-BJDJZHNGSA-N 0.000 description 2
- GDUZTEQRAOXYJS-SRVKXCTJSA-N Ser-Phe-Asn Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CO)N GDUZTEQRAOXYJS-SRVKXCTJSA-N 0.000 description 2
- NADLKBTYNKUJEP-KATARQTJSA-N Ser-Thr-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O NADLKBTYNKUJEP-KATARQTJSA-N 0.000 description 2
- SNXUIBACCONSOH-BWBBJGPYSA-N Ser-Thr-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CO)C(O)=O SNXUIBACCONSOH-BWBBJGPYSA-N 0.000 description 2
- FHXGMDRKJHKLKW-QWRGUYRKSA-N Ser-Tyr-Gly Chemical compound OC[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CC1=CC=C(O)C=C1 FHXGMDRKJHKLKW-QWRGUYRKSA-N 0.000 description 2
- BEBVVQPDSHHWQL-NRPADANISA-N Ser-Val-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O BEBVVQPDSHHWQL-NRPADANISA-N 0.000 description 2
- TYVAWPFQYFPSBR-BFHQHQDPSA-N Thr-Ala-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)NCC(O)=O TYVAWPFQYFPSBR-BFHQHQDPSA-N 0.000 description 2
- GLQFKOVWXPPFTP-VEVYYDQMSA-N Thr-Arg-Asp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(O)=O GLQFKOVWXPPFTP-VEVYYDQMSA-N 0.000 description 2
- MQBTXMPQNCGSSZ-OSUNSFLBSA-N Thr-Arg-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)[C@@H](C)O)CCCN=C(N)N MQBTXMPQNCGSSZ-OSUNSFLBSA-N 0.000 description 2
- JNQZPAWOPBZGIX-RCWTZXSCSA-N Thr-Arg-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)[C@@H](C)O)CCCN=C(N)N JNQZPAWOPBZGIX-RCWTZXSCSA-N 0.000 description 2
- LXWZOMSOUAMOIA-JIOCBJNQSA-N Thr-Asn-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N1CCC[C@@H]1C(=O)O)N)O LXWZOMSOUAMOIA-JIOCBJNQSA-N 0.000 description 2
- VGYBYGQXZJDZJU-XQXXSGGOSA-N Thr-Glu-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O VGYBYGQXZJDZJU-XQXXSGGOSA-N 0.000 description 2
- YZUWGFXVVZQJEI-PMVVWTBXSA-N Thr-Gly-His Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N)O YZUWGFXVVZQJEI-PMVVWTBXSA-N 0.000 description 2
- SXAGUVRFGJSFKC-ZEILLAHLSA-N Thr-His-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SXAGUVRFGJSFKC-ZEILLAHLSA-N 0.000 description 2
- YJCVECXVYHZOBK-KNZXXDILSA-N Thr-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H]([C@@H](C)O)N YJCVECXVYHZOBK-KNZXXDILSA-N 0.000 description 2
- KZURUCDWKDEAFZ-XVSYOHENSA-N Thr-Phe-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)O KZURUCDWKDEAFZ-XVSYOHENSA-N 0.000 description 2
- MROIJTGJGIDEEJ-RCWTZXSCSA-N Thr-Pro-Pro Chemical compound C[C@@H](O)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 MROIJTGJGIDEEJ-RCWTZXSCSA-N 0.000 description 2
- SGAOHNPSEPVAFP-ZDLURKLDSA-N Thr-Ser-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)NCC(O)=O SGAOHNPSEPVAFP-ZDLURKLDSA-N 0.000 description 2
- MYNYCUXMIIWUNW-IEGACIPQSA-N Thr-Trp-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CCCCN)C(=O)O)N)O MYNYCUXMIIWUNW-IEGACIPQSA-N 0.000 description 2
- ABCLYRRGTZNIFU-BWAGICSOSA-N Thr-Tyr-His Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N)O ABCLYRRGTZNIFU-BWAGICSOSA-N 0.000 description 2
- BXKWZPXTTSCOMX-AQZXSJQPSA-N Trp-Asn-Thr Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O BXKWZPXTTSCOMX-AQZXSJQPSA-N 0.000 description 2
- BEIGSKUPTIFYRZ-SRVKXCTJSA-N Tyr-Asp-Asp Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC(=O)O)C(=O)O)N)O BEIGSKUPTIFYRZ-SRVKXCTJSA-N 0.000 description 2
- CKHQKYHIZCRTAP-SOUVJXGZSA-N Tyr-Gln-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC2=CC=C(C=C2)O)N)C(=O)O CKHQKYHIZCRTAP-SOUVJXGZSA-N 0.000 description 2
- LOOCQRRBKZTPKO-AVGNSLFASA-N Tyr-Glu-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 LOOCQRRBKZTPKO-AVGNSLFASA-N 0.000 description 2
- FNWGDMZVYBVAGJ-XEGUGMAKSA-N Tyr-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CC1=CC=C(C=C1)O)N FNWGDMZVYBVAGJ-XEGUGMAKSA-N 0.000 description 2
- BYAKMYBZADCNMN-JYJNAYRXSA-N Tyr-Lys-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(O)=O BYAKMYBZADCNMN-JYJNAYRXSA-N 0.000 description 2
- LMKKMCGTDANZTR-BZSNNMDCSA-N Tyr-Phe-Asp Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC(O)=O)C(O)=O)C1=CC=C(O)C=C1 LMKKMCGTDANZTR-BZSNNMDCSA-N 0.000 description 2
- QKXAEWMHAAVVGS-KKUMJFAQSA-N Tyr-Pro-Glu Chemical compound N[C@@H](Cc1ccc(O)cc1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O QKXAEWMHAAVVGS-KKUMJFAQSA-N 0.000 description 2
- GAKBTSMAPGLQFA-JNPHEJMOSA-N Tyr-Thr-Tyr Chemical compound C([C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 GAKBTSMAPGLQFA-JNPHEJMOSA-N 0.000 description 2
- VKYDVKAKGDNZED-STECZYCISA-N Tyr-Val-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC1=CC=C(C=C1)O)N VKYDVKAKGDNZED-STECZYCISA-N 0.000 description 2
- FZSPNKUFROZBSG-ZKWXMUAHSA-N Val-Ala-Asp Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O FZSPNKUFROZBSG-ZKWXMUAHSA-N 0.000 description 2
- CFSSLXZJEMERJY-NRPADANISA-N Val-Gln-Ala Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O CFSSLXZJEMERJY-NRPADANISA-N 0.000 description 2
- URIRWLJVWHYLET-ONGXEEELSA-N Val-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)C(C)C URIRWLJVWHYLET-ONGXEEELSA-N 0.000 description 2
- ZHQWPWQNVRCXAX-XQQFMLRXSA-N Val-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](C(C)C)N ZHQWPWQNVRCXAX-XQQFMLRXSA-N 0.000 description 2
- BTWMICVCQLKKNR-DCAQKATOSA-N Val-Leu-Ser Chemical compound CC(C)[C@H]([NH3+])C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C([O-])=O BTWMICVCQLKKNR-DCAQKATOSA-N 0.000 description 2
- DEGUERSKQBRZMZ-FXQIFTODSA-N Val-Ser-Ala Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O DEGUERSKQBRZMZ-FXQIFTODSA-N 0.000 description 2
- UGFMVXRXULGLNO-XPUUQOCRSA-N Val-Ser-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O UGFMVXRXULGLNO-XPUUQOCRSA-N 0.000 description 2
- PGQUDQYHWICSAB-NAKRPEOUSA-N Val-Ser-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](C(C)C)N PGQUDQYHWICSAB-NAKRPEOUSA-N 0.000 description 2
- 108010070944 alanylhistidine Proteins 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 239000004202 carbamide Substances 0.000 description 2
- 125000004432 carbon atom Chemical group C* 0.000 description 2
- 239000013592 cell lysate Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 235000013351 cheese Nutrition 0.000 description 2
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 2
- 239000006071 cream Substances 0.000 description 2
- 239000012228 culture supernatant Substances 0.000 description 2
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 2
- 235000018417 cysteine Nutrition 0.000 description 2
- 239000003599 detergent Substances 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 238000003912 environmental pollution Methods 0.000 description 2
- 108010063718 gamma-glutamylaspartic acid Proteins 0.000 description 2
- 108010082286 glycyl-seryl-alanine Proteins 0.000 description 2
- 108010010147 glycylglutamine Proteins 0.000 description 2
- 108010037850 glycylvaline Proteins 0.000 description 2
- 108010018006 histidylserine Proteins 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 230000002779 inactivation Effects 0.000 description 2
- ZXEKIIBDNHEJCQ-UHFFFAOYSA-N isobutanol Chemical compound CC(C)CO ZXEKIIBDNHEJCQ-UHFFFAOYSA-N 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 108010044374 isoleucyl-tyrosine Proteins 0.000 description 2
- 108010044056 leucyl-phenylalanine Proteins 0.000 description 2
- 108010045397 lysyl-tyrosyl-lysine Proteins 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 108010085203 methionylmethionine Proteins 0.000 description 2
- 201000000050 myeloid neoplasm Diseases 0.000 description 2
- 108010073101 phenylalanylleucine Proteins 0.000 description 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 235000019419 proteases Nutrition 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 108091008146 restriction endonucleases Proteins 0.000 description 2
- 229920002477 rna polymer Polymers 0.000 description 2
- 108010026333 seryl-proline Proteins 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000004094 surface-active agent Substances 0.000 description 2
- 238000001890 transfection Methods 0.000 description 2
- 150000003626 triacylglycerols Chemical class 0.000 description 2
- 108010084932 tryptophyl-proline Proteins 0.000 description 2
- 108010035534 tyrosyl-leucyl-alanine Proteins 0.000 description 2
- 108010051110 tyrosyl-lysine Proteins 0.000 description 2
- 108010003137 tyrosyltyrosine Proteins 0.000 description 2
- 108010073969 valyllysine Proteins 0.000 description 2
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 1
- VZSRBBMJRBPUNF-UHFFFAOYSA-N 2-(2,3-dihydro-1H-inden-2-ylamino)-N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]pyrimidine-5-carboxamide Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C(=O)NCCC(N1CC2=C(CC1)NN=N2)=O VZSRBBMJRBPUNF-UHFFFAOYSA-N 0.000 description 1
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- YLZOPXRUQYQQID-UHFFFAOYSA-N 3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)-1-[4-[2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidin-5-yl]piperazin-1-yl]propan-1-one Chemical compound N1N=NC=2CN(CCC=21)CCC(=O)N1CCN(CC1)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F YLZOPXRUQYQQID-UHFFFAOYSA-N 0.000 description 1
- 244000215068 Acacia senegal Species 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 239000004382 Amylase Substances 0.000 description 1
- 102000013142 Amylases Human genes 0.000 description 1
- 108010065511 Amylases Proteins 0.000 description 1
- 108700042778 Antimicrobial Peptides Proteins 0.000 description 1
- 102000044503 Antimicrobial Peptides Human genes 0.000 description 1
- 101001073212 Arabidopsis thaliana Peroxidase 33 Proteins 0.000 description 1
- UBCPNBUIQNMDNH-NAKRPEOUSA-N Arg-Ile-Ala Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O UBCPNBUIQNMDNH-NAKRPEOUSA-N 0.000 description 1
- KPSHWSWFPUDEGF-FXQIFTODSA-N Asp-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CC(O)=O KPSHWSWFPUDEGF-FXQIFTODSA-N 0.000 description 1
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 1
- 238000009010 Bradford assay Methods 0.000 description 1
- 101100505161 Caenorhabditis elegans mel-32 gene Proteins 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 108010031797 Candida antarctica lipase B Proteins 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 238000001712 DNA sequencing Methods 0.000 description 1
- 241001198387 Escherichia coli BL21(DE3) Species 0.000 description 1
- SHERTACNJPYHAR-ACZMJKKPSA-N Gln-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCC(N)=O SHERTACNJPYHAR-ACZMJKKPSA-N 0.000 description 1
- SSHIXEILTLPAQT-WHFBIAKZSA-N Gln-Asp Chemical compound NC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(O)=O SSHIXEILTLPAQT-WHFBIAKZSA-N 0.000 description 1
- RJIVPOXLQFJRTG-LURJTMIESA-N Gly-Arg-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)CN)CCCN=C(N)N RJIVPOXLQFJRTG-LURJTMIESA-N 0.000 description 1
- OVSKVOOUFAKODB-UWVGGRQHSA-N Gly-Arg-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CCCN=C(N)N OVSKVOOUFAKODB-UWVGGRQHSA-N 0.000 description 1
- NNCSJUBVFBDDLC-YUMQZZPRSA-N Gly-Leu-Ser Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O NNCSJUBVFBDDLC-YUMQZZPRSA-N 0.000 description 1
- LOEANKRDMMVOGZ-YUMQZZPRSA-N Gly-Lys-Asp Chemical compound NCCCC[C@H](NC(=O)CN)C(=O)N[C@@H](CC(O)=O)C(O)=O LOEANKRDMMVOGZ-YUMQZZPRSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 229920000209 Hexadimethrine bromide Polymers 0.000 description 1
- 101001123325 Homo sapiens Peroxisome proliferator-activated receptor gamma coactivator 1-beta Proteins 0.000 description 1
- JHNJNTMTZHEDLJ-NAKRPEOUSA-N Ile-Ser-Arg Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O JHNJNTMTZHEDLJ-NAKRPEOUSA-N 0.000 description 1
- VGSPNSSCMOHRRR-BJDJZHNGSA-N Ile-Ser-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)O)N VGSPNSSCMOHRRR-BJDJZHNGSA-N 0.000 description 1
- 241000235058 Komagataella pastoris Species 0.000 description 1
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 1
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 1
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 1
- SENJXOPIZNYLHU-UHFFFAOYSA-N L-leucyl-L-arginine Natural products CC(C)CC(N)C(=O)NC(C(O)=O)CCCN=C(N)N SENJXOPIZNYLHU-UHFFFAOYSA-N 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- IZPVWNSAVUQBGP-CIUDSAMLSA-N Leu-Ser-Asp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O IZPVWNSAVUQBGP-CIUDSAMLSA-N 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 241001661345 Moesziomyces antarcticus Species 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- NIPNSKYNPDTRPC-UHFFFAOYSA-N N-[2-oxo-2-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)ethyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 NIPNSKYNPDTRPC-UHFFFAOYSA-N 0.000 description 1
- 125000001429 N-terminal alpha-amino-acid group Chemical group 0.000 description 1
- 206010029719 Nonspecific reaction Diseases 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 102100028961 Peroxisome proliferator-activated receptor gamma coactivator 1-beta Human genes 0.000 description 1
- 239000004372 Polyvinyl alcohol Substances 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 235000019484 Rapeseed oil Nutrition 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 240000000528 Ricinus communis Species 0.000 description 1
- 235000004443 Ricinus communis Nutrition 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 241000293869 Salmonella enterica subsp. enterica serovar Typhimurium Species 0.000 description 1
- GXXTUIUYTWGPMV-FXQIFTODSA-N Ser-Arg-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(O)=O GXXTUIUYTWGPMV-FXQIFTODSA-N 0.000 description 1
- WSTIOCFMWXNOCX-YUMQZZPRSA-N Ser-Gly-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CO)N WSTIOCFMWXNOCX-YUMQZZPRSA-N 0.000 description 1
- BXLYSRPHVMCOPS-ACZMJKKPSA-N Ser-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](N)CO BXLYSRPHVMCOPS-ACZMJKKPSA-N 0.000 description 1
- MQQBBLVOUUJKLH-HJPIBITLSA-N Ser-Ile-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O MQQBBLVOUUJKLH-HJPIBITLSA-N 0.000 description 1
- XZKQVQKUZMAADP-IMJSIDKUSA-N Ser-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(O)=O XZKQVQKUZMAADP-IMJSIDKUSA-N 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- 241000256248 Spodoptera Species 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- 241000187747 Streptomyces Species 0.000 description 1
- DKGAVHZHDRPRBM-UHFFFAOYSA-N Tert-Butanol Chemical compound CC(C)(C)O DKGAVHZHDRPRBM-UHFFFAOYSA-N 0.000 description 1
- CEXFELBFVHLYDZ-XGEHTFHBSA-N Thr-Arg-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(O)=O CEXFELBFVHLYDZ-XGEHTFHBSA-N 0.000 description 1
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 1
- 239000004473 Threonine Substances 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 244000098338 Triticum aestivum Species 0.000 description 1
- RRVUOLRWIZXBRQ-IHPCNDPISA-N Trp-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N RRVUOLRWIZXBRQ-IHPCNDPISA-N 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- CXWJFWAZIVWBOS-XQQFMLRXSA-N Val-Lys-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N1CCC[C@@H]1C(=O)O)N CXWJFWAZIVWBOS-XQQFMLRXSA-N 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- 238000005411 Van der Waals force Methods 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 239000000205 acacia gum Substances 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 125000003172 aldehyde group Chemical group 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- 235000019418 amylase Nutrition 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 235000009582 asparagine Nutrition 0.000 description 1
- 229960001230 asparagine Drugs 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000001588 bifunctional effect Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 235000014121 butter Nutrition 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- HJMZMZRCABDKKV-UHFFFAOYSA-N carbonocyanidic acid Chemical compound OC(=O)C#N HJMZMZRCABDKKV-UHFFFAOYSA-N 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000006037 cell lysis Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000000975 co-precipitation Methods 0.000 description 1
- 229940110456 cocoa butter Drugs 0.000 description 1
- 235000019868 cocoa butter Nutrition 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000008162 cooking oil Substances 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 102000038379 digestive enzymes Human genes 0.000 description 1
- 108091007734 digestive enzymes Proteins 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 230000009881 electrostatic interaction Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 125000001153 fluoro group Chemical group F* 0.000 description 1
- 125000002485 formyl group Chemical class [H]C(*)=O 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 238000001641 gel filtration chromatography Methods 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 235000004554 glutamine Nutrition 0.000 description 1
- 125000005456 glyceride group Chemical group 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 230000005661 hydrophobic surface Effects 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 229960000310 isoleucine Drugs 0.000 description 1
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 1
- FZWBNHMXJMCXLU-BLAUPYHCSA-N isomaltotriose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O)O1 FZWBNHMXJMCXLU-BLAUPYHCSA-N 0.000 description 1
- 229960004592 isopropanol Drugs 0.000 description 1
- 108010000761 leucylarginine Proteins 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- 108010003700 lysyl aspartic acid Proteins 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 201000001441 melanoma Diseases 0.000 description 1
- 238000011034 membrane dialysis Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 150000004702 methyl esters Chemical class 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 235000021243 milk fat Nutrition 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 210000001819 pancreatic juice Anatomy 0.000 description 1
- 239000010893 paper waste Substances 0.000 description 1
- NFEQUGKCQWAGLY-UAAVROCESA-N parasin i Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CCCCN)NC(=O)CNC(=O)CNC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCCCN)NC(=O)CNC(=O)[C@H](CCCNC(N)=N)NC(=O)CNC(=O)[C@@H](N)CCCCN NFEQUGKCQWAGLY-UAAVROCESA-N 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 235000015277 pork Nutrition 0.000 description 1
- 210000001236 prokaryotic cell Anatomy 0.000 description 1
- 108010015796 prolylisoleucine Proteins 0.000 description 1
- 230000006920 protein precipitation Effects 0.000 description 1
- 239000012460 protein solution Substances 0.000 description 1
- 210000001938 protoplast Anatomy 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000003362 replicative effect Effects 0.000 description 1
- 230000002207 retinal effect Effects 0.000 description 1
- 230000001177 retroviral effect Effects 0.000 description 1
- 230000005070 ripening Effects 0.000 description 1
- 235000004400 serine Nutrition 0.000 description 1
- 238000002741 site-directed mutagenesis Methods 0.000 description 1
- 239000000344 soap Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 239000003549 soybean oil Substances 0.000 description 1
- 235000012424 soybean oil Nutrition 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 239000003760 tallow Substances 0.000 description 1
- 235000008521 threonine Nutrition 0.000 description 1
- 238000010361 transduction Methods 0.000 description 1
- 230000026683 transduction Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 235000002374 tyrosine Nutrition 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 229920002554 vinyl polymer Polymers 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 239000011534 wash buffer Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000002351 wastewater Substances 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
- 210000005253 yeast cell Anatomy 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/001—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof by chemical synthesis
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/463—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from amphibians
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4723—Cationic antimicrobial peptides, e.g. defensins
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/52—Genes encoding for enzymes or proenzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/62—DNA sequences coding for fusion proteins
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
- C12N9/18—Carboxylic ester hydrolases (3.1.1)
- C12N9/20—Triglyceride splitting, e.g. by means of lipase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6436—Fatty acid esters
- C12P7/6445—Glycerides
- C12P7/6458—Glycerides by transesterification, e.g. interesterification, ester interchange, alcoholysis or acidolysis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6436—Fatty acid esters
- C12P7/649—Biodiesel, i.e. fatty acid alkyl esters
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y301/00—Hydrolases acting on ester bonds (3.1)
- C12Y301/01—Carboxylic ester hydrolases (3.1.1)
- C12Y301/01003—Triacylglycerol lipase (3.1.1.3)
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
- C07K2319/21—Fusion polypeptide containing a tag with affinity for a non-protein ligand containing a His-tag
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- Biophysics (AREA)
- General Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Gastroenterology & Hepatology (AREA)
- Biomedical Technology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Toxicology (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Abstract
본 발명은 리파아제 활성이 증대된 양친매성 펩타이드(amphipathic peptide) 및 리파아제가 연결된 양친매성 펩타이드-리파아제 결합체, 상기 결합체를 코딩하는 폴리뉴클레오티드, 상기 폴리뉴클레오티드를 포함하는 발현벡터, 상기 발현벡터가 도입된 형질전환체, 상기 결합체의 제조방법, 상기 결합체를 이용한 지질의 분해방법 및 상기 리파아제를 이용한 바이오디젤의 생산방법에 관한 것이다.
Description
본 발명은 리파아제 활성이 증대된 양친매성 펩타이드(amphipathic peptide) 및 리파아제가 연결된 양친매성 펩타이드-리파아제 결합체, 상기 결합체를 코딩하는 폴리뉴클레오티드, 상기 폴리뉴클레오티드를 포함하는 발현벡터, 상기 발현벡터가 도입된 형질전환체, 상기 결합체의 제조방법, 상기 결합체를 이용한 지질의 분해방법 및 상기 리파아제를 이용한 바이오디젤의 생산방법에 관한 것이다.
리파아제(lipase; triacylglycerol acylhydrolase, EC3.1.1.3)는 트리글리세라이드를 지방산과 글리세라이드 혹은 글리세롤로 가수분해하는 효소로서, 아밀라제(amylase) 및 프로테아제(protease)와 함께 3대 소화효소의 주축을 이루고, 동물, 식물 및 미생물 등의 자연계에 광범위하게 분포한다.
상기 리파아제는 생체 내에서 지방대사의 중요한 역할을 할 뿐 아니라, 치즈 풍미 증진, 야채 발효시 유리지방산 증가, 육류 숙성시 향기 증진 및 어류 지방 분해에도 작용하고, 고가의 순수광학 이성질체 합성에도 사용될 수 있는 등의 폭넓은 활용범위를 갖고 있는 것으로 알려져 있다. 또한, 산업적으로도 널리 활용되고 있는데, 구체적으로, 낙농산업 분야에서는 유지방 분해를 통한 치즈의 제조, 버터나 크림의 지방분해를 위해 사용되고, 세제산업 분야에서는 세탁 또는 세척세제의 제조, 환경오염 폐수 감소, 반응온도 감소 등에 사용되며, 유화학산업 분야에서는 불포화 지방산 생산, 비누제조, 싼 팜유를 이용한 코코아 버터의 생산 등에 활용되고, 제지산업 분야에서는 나무의 송진 또는 수지 제거, 폐지의 잉크제거 등에 활용되며, 제약산업 분야에서는 R, S- 광학이성질체 구분 합성, 라세믹 혼합물의 분리, 약제조 등에 이용되고, 화장품산업 분야에서는 왁스를 비롯한 피부용품, 선탠크림, 목욕용품 등의 제조에 이용되며, 에너지산업 분야에서는 식물성 기름으로부터 바이오디젤의 생산에 활용되고 있다.
특히, 신재생 에너지로 평가받고 있는 바이오디젤은 현재 국내외에서 화학촉매를 이용한 방법으로 생산되고 있지만, 다량의 유기 용매 사용, 환경처리 비용, 고온의 반응 조건에 따른 에너지 과다사용 등의 문제로 인한 높은 생산 단가 때문에 상업화가 어려워 효소인 리파아제를 촉매로 이용하는 공정이 주목받고 있다. 상기 공정을 이용할 경우, 낮은 반응온도에 따른 에너지 절약과 부산물인 글리세롤을 이용한 추가 수익 발생을 통하여, 생산비용을 낮출 수 있고, 환경오염 문제가 거의 없다는 장점이 있는 반면, 효소의 안정성이 약하고, 비용대비 효율이 낮다는 단점이 있었다. 상기 단점을 해결하기 위하여 효소를 고정화하는 방법이 개발되었으나, 이는 효소의 특징을 개선한다는 직접적인 연구 방법이기 보다 간접적인 해결책을 제시한 것이기 때문에, 직접적인 효소의 활성을 증대시켜 산업적 이용가치를 증가시키는 방법을 개발하려는 연구가 활발히 진행되었다.
예를 들어, 리파아제의 효율적 활용을 위하여 저렴하면서도 강력한 활성의 리파아제를 개발하기보다는 기존의 리파아제의 반응조건을 바꾸거나, 효소 고정화와 같은 간접적 방법을 통해서 문제를 해결하거나 또는 메타제노믹스를 통해 새로운 리파아제를 찾으려는 연구가 수행되었으나, 별다른 성과를 얻지 못하였다. 또한, 다양한 리파아제의 X-ray 구조를 분석하고, 효소 작용부위와 그 주변의 아미노산 치환하여 효소활성, 기질 특이성, 내열성, 프로테아제 안정성을 증가시키는 연구, 특히 위치지정 돌연변이 방법(site-directed mutagenesis)과 같은 단백질 공학을 사용하여 리파아제를 개량하려는 연구가 많이 진행되어 왔으나, 리파아제의 활성이 부분적으로 증가되는 결과만을 얻을 수 있었다.
이러한 배경 하에서, 본 발명자들은 리파아제의 기질이 불용성이기 때문에 상기 기질에 대한 리파아제의 접근성이 낮다는 점에 착안하여, 기존의 리파아제에 친수기와 소수기를 가지는 양친매성(amphiphacity)의 펩타이드를 융합시켜, 양친매성 펩타이드 및 리파아제를 포함하는 융합 폴리펩타이드를 제작하였고, 상기 융합 폴리펩타이드를 사용할 경우, 기질에 대한 접근성, 결합성 및 반응성이 향상됨을 확인하고, 본 발명을 완성하였다.
본 발명의 하나의 목적은 양친매성 펩타이드(amphipathic peptide) 및 리파아제가 연결된 양친매성 펩타이드-리파아제 결합체를 제공하는 것이다.
본 발명의 다른 목적은 상기 결합체를 코딩하는 폴리뉴클레오티드, 상기 폴리뉴클레오티드를 포함하는 발현벡터 및 상기 발현벡터가 도입된 형질전환체를 제공하는 것이다.
본 발명의 또 다른 목적은 상기 양친매성 펩타이드-리파아제 결합체의 제조방법을 제공하는 것이다.
본 발명의 또 다른 목적은 상기 양친매성 펩타이드-리파아제 결합체를 이용한 지질의 분해방법을 제공하는 것이다.
본 발명의 또 다른 목적은 상기 양친매성 펩타이드-리파아제 결합체를 이용한 바이오디젤의 생산방법을 제공하는 것이다.
상기의 목적을 달성하기 위한 하나의 양태로서, 본 발명은 양친매성 펩타이드(amphipathic peptide) 및 리파아제가 연결된 양친매성 펩타이드-리파아제 결합체를 제공한다.
본 발명에서 용어, "양친매성 펩타이드"란 극성 아미노산과 비극성 아미노산을 포함하고, 상기 극성 아미노산과 비극성 아미노산이 부분적으로 서로 밀집되어 극성 부분과 비극성 부분을 나타내는 펩타이드를 의미하는데, 상기 극성 아미노산으로는 시스테인, 글루타민, 트레오닌, 타이로신, 세린 및 아스파라긴을 들 수 있고, 상기 비극성 아미노산으로는 페닐알라닌, 트립토판, 메티오닌, 프롤린, 발린, 이소류신, 류신, 글리신 및 알라닌을 들 수 있다. 본 발명에서 상기 양친매성 펩타이드는 리파아제와 그의 기질인 지방성분 간의 결합 및 반응성을 증가시키는 매체로서 사용될 수 있는데, 아미노산 서열상 극성 아미노산과 비극성 아미노산이 서로 밀집된 형태일 수 있고, 아미노산 서열상 극성 아미노산과 비극성 아미노산이 서로 밀집되어 있지는 않으나, 상기 아미노산을 포함하는 펩타이드가 3차원 구조를 형성할 때, 상기 3차원 구조에 의하여 극성 아미노산과 비극성 아미노산이 서로 밀집되는 형태일 수도 있으나, 특별히 이에 제한되지는 않는다.
본 발명의 목적상 상기 양친매성 펩타이드는 리파아제와 결합하여 리파아제의 기질 접근성 또는 반응성을 향상시킬 수 있는 펩타이드라면 제한없이 포함하며, 그 예로 부포린 IIb(Buforin IIb, 서열번호 1), B0(서열번호 2), 파라신 I(Paracin I, 서열번호 3), NKC(서열번호 4), NRC(서열번호 5)일 수 있으나, 이에 제한되지 않는다. 또한, 상기 양친매성 펩타이드는 리파아제와 결합하여 리파아제의 기질 접근성 또는 반응성을 향상시킬 수 있는 펩타이드라면 특별히 그 길이에 제한되지 않는다. 본 발명의 일 실시예에서는 부포린 IIb, B0, 파라신 I, NKC 및 NRC를 대표적인 양친매성 펩타이드로 사용하였다.
상기 양친매성 펩타이드는 서열번호 1 내지 5로 구성되는 군으로부터 선택되는 아미노산 서열을 가지는 펩타이드뿐만 아니라, 상기 서열과 70% 이상, 바람직하게는 80% 이상, 더욱 바람직하게는 90% 이상, 보다 더욱 바람직하게는 95% 이상, 가장 바람직하게는 98% 이상의 유사성을 나타내는 아미노산 서열로서 실질적으로 리파아제의 활성을 증진시킬 수 있는 생물학적 활성을 갖는 단백질이라면 모두 포함한다. 또한, 이러한 유사성을 갖는 서열로서 실질적으로 상기 양친매성 펩타이드와 동일하거나 상응하는 생물학적 활성을 갖는 아미노산 서열이라면, 일부 서열이 결실, 변형, 치환 또는 부가된 아미노산 서열을 갖는 펩타이드 변이체도 본 발명의 범위 내에 포함됨은 자명하다.
본 발명에서 용어, "리파아제(lipase)"란 지방을 분해하는 효소를 의미하는데, 구체적으로 중성지방에 작용하여 트라이아실글리세리드에서 다이아실글리세리드와 모노아실글리세리드를 거쳐서 지방산과 글리세린으로 분해하는 효소를 의미한다. 대체로 동물의 췌장액에 풍부하게 포함되어 있고, 식물에서는 밀, 아주까리, 콩 등의 종자에 포함되어 있다. 상기 리파아제는 지방을 분해하는 생물학적 활성을 지닌 효소라면 그 종류가 특별히 제한되지 않으나, 그 예로 슈도모나스 플루오레슨스(Pseudomonas fluorescens) 유래의 리파아제 TliA(thermostable lipase) 또는 포토박테리움 리포리티쿰(Photobacterium lipolyticum) 유래의 리파아제 M37일 수 있으나, 이에 제한되지 않는다. 상기 리파아제에 관한 정보는 미국 국립보건원 GenBank와 같은 공지의 데이터베이스로부터 얻을 수 있으며, 그 예로 Accession Number가 AAD09856인 TliA 또는 AAS78630인 M37일 수 있으나, 이에 제한되지 않는다. 상기 TliA 리파아제는 서열번호 6으로 기재된 아미노산 서열로 정의되는 리파아제이거나, M37은 서열번호 7로 기재된 아미노산 서열로 정의되는 리파아제일 수 있으나, 이에 제한되지 않는다. 또한, 상기 리파아제는 서열번호 6 또는 서열번호 7로 기재된 아미노산 서열로 정의되는 리파아제뿐만 아니라, 상기 서열과 70% 이상, 바람직하게는 80% 이상, 더욱 바람직하게는 90% 이상, 보다 더욱 바람직하게는 95% 이상, 가장 바람직하게는 98% 이상의 유사성을 나타내는 아미노산 서열로서 실질적으로 지질을 분해하는 생물학적 활성을 갖는 단백질이라면 모두 포함한다. 또한, 이러한 유사성을 갖는 서열로서 실질적으로 상기 리파아제와 동일하거나 상응하는 생물학적 활성을 갖는 아미노산 서열이라면, 일부 서열이 결실, 변형, 치환 또는 부가된 아미노산 서열을 갖는 단백질 변이체도 본 발명의 범위 내에 포함됨은 자명하다.
본 발명에서 용어, "양친매성 펩타이드-리파아제 결합체"는 양친매성 펩타이드가 리파아제의 생물학적 활성을 저해시키지 않도록 리파아제에 연결된 형태를 의미한다. 상기 양친매성 펩타이드-리파아제 결합체는 화학적으로 연결되거나, 두 종류 이상의 폴리펩타이드가 효소작용에 의하여 연결되거나, 또는 유전자의 조작을 통해 두 종류 이상의 상이한 폴리펩타이드가 하나의 폴리펩타이드로 발현된 형태로 연결될 수 있으나, 이에 제한되지 않는다.
상기 양친매성 펩타이드와 리파아제는 양친매성 펩타이드와 리파아제가 직접 연결된 형태일 수도 있고, 링커(linker)에 의해 연결된 형태일 수 있으나, 이에 제한되지 않는다.
본 발명에서 용어, "링커(linker)"란 기본적으로는 두 개의 서로 다른 융합 파트너(예를 들어, 생물학적 고분자 등)를 수소결합, 정전기적 상호작용, 반데르발스력, 이황화 결합, 염 브릿지, 소수성 상호작용, 공유결합 등을 이용하여 연결할 수 있는 연결체를 의미한다. 바람직하게는 생리학적 조건 또는 다른 표준 펩타이드 조건(예를 들면, 펩타이드 정제 조건, 펩타이드 저장 조건)하에서 적어도 하나의 이황화 결합에 참여할 수 있는 적어도 하나의 시스테인을 가질 수 있으며, 단순히 각각의 융합 파트너를 연결하는 역할 이외에도, 융합 파트너 사이에 일정한 크기의 간격을 부여하는 역할을 수행하거나 또는 융합체에 유연성 또는 강직성을 제공하는 힌지(hinge)의 역할을 수행할 수도 있다. 상기 링커는 비펩타이드 링커 또는 펩타이드 링커일 수 있으며, 펩티드 결합, 이황화 결합 등에 의해서 직접 연결되는 것도 포함된다.
본 발명에서 용어, "비펩타이드 링커"는 반복 단위가 2개 이상 결합된 생체적합성 링커를 의미하며, 상기 반복 단위들은 펩타이드 결합이 아닌 임의의 공유결합을 통해 서로 연결된다.
본 발명에 사용가능한 비펩타이드 링커는 폴리에틸렌 글리콜(polyethylene glycol; PEG) 단독 중합체, 폴리프로필렌 글리콜 단독 중합체, 에틸렌 글리콜-프로필렌 글리콜 공중합체, 폴리옥시 에틸화 폴리올, 폴리비닐 알콜, 폴리사카라이드, 덱스트란, 폴리비닐 에틸 에테르, 생분해성 고분자, 지질 중합체, 키틴류, 히아루론산 또는 이들의 조합일 수 있다. 바람직하게는 폴리에틸렌 글리콜 단독 중합체이다. 당해 분야에 이미 알려진 이들의 유도체 및 당해 분야의 기술 수준에서 용이하게 제조할 수 있는 유도체들도 본 발명의 범위에 포함된다. 더욱 바람직하게는 분자량 1 내지 5 kDa의 분자량인 폴리에틸렌 글리콜 단독 중합체이며, 가장 바람직하게는 3.4 kDa 정도이며, 양 말단에 양기능성 알데히드(bifunctional aldehyde) 형태로 폴리펩타이드 아미노 말단의 아민그룹과 결합하여, 양친매성 펩타이드와 리파아제를 서로 연결시킬 수 있다. 특히, 양 말단에 반응 알데히드 그룹의 반응기를 갖는 경우, 비특이적 반응을 최소화하는 데 효과적이다.
또한, 상기 양친매성 펩타이드와 리파아제의 결합 형태는 특별히 이에 제한되지 않으나, 리파아제의 N-말단에 양친매성 펩타이드가 결합한 형태가 될 수 있고, 상기 리파아제의 C-말단에 양친매성 펩타이드가 결합한 형태가 될 수도 있으며, 상기 리파아제의 N-말단과 C-말단에 양친매성 펩타이드가 각각 결합한 형태가 될 수도 있다. 예를 들어, 리파아제 TliA의 N-말단에 부포린 IIb, B0, 파라신 I, NKC 또는 NRC가 융합된 형태(서열번호 8 내지 12), 리파아제 M37의 N-말단에 부포린 IIb, B0, 파라신 I, NKC 또는 NRC가 융합된 형태(서열번호 13 내지 17), 리파아제 TliA의 C-말단에 부포린 IIb, B0, 파라신 I, NKC 또는 NRC가 융합된 형태(서열번호 18 내지 22), 리파아제 M37의 C-말단에 부포린 IIb, B0, 파라신 I, NKC 또는 NRC가 융합된 형태(서열번호 23 내지 27), 리파아제 TliA의 N-말단과 C-말단에 부포린 IIb, B0, 파라신 I, NKC 또는 NRC가 융합된 형태(서열번호 28 내지 32), 리파아제 M37의 N-말단과 C-말단에 부포린 IIb, B0, 파라신 I, NKC 또는 NRC가 융합된 형태(서열번호 33 내지 37) 등이 될 수 있다.
본 발명의 일 실시예에서는, 부포린 IIb, B0, 파라신 I, NKC 또는 NRC를 양친매성 펩타이드로 사용하였고, 리파아제로 TliA 또는 M37을 사용하였다. 상기 양친매성 펩타이드를 리파아제의 N-말단에 연결시킨 양친매성 펩타이드-리파아제 결합체를 제조하였고, 상기 양친매성 펩타이드-리파아제가 리파아제의 지질 분해 활성을 최대 10배까지 증대시킴을 확인하여(도 4 내지 7), 본 발명의 양친매성 펩타이드를 연결시킨 리파아제가 뛰어난 지질 분해 활성을 나타냄을 확인하여, 지질 분해가 필요한 다양한 분야에 사용될 수 있는 경제적이면서도 뛰어난 리파아제임을 확인하였다. 또한, 양친매성 펩타이드를 리파아제에 결합시킨 경우, 지질 입자에 대한 접근성 또한 향상되는 결과를 보였다(도 8). 아울러, 본 발명의 대표적인 양친매성 펩타이드-리파아제 결합체인 NKC-M37 리파아제의 경우, 바이오디젤의 생산에 사용되는 메탄올의 존재하에서도 야생형 리파아제인 M37에 비하여 뛰어난 안정성 및 지질분해활성을 보임으로써(도 9 및 10), 본 발명의 양친매성 펩타이드-리파아제 결합체가 바이오디젤의 생산에 유용하게 사용될 수 있음을 시사하였다.
또 하나의 양태로서 본 발명은 상기 결합체를 코딩하는 폴리뉴클레오티드, 상기 폴리뉴클레오티드를 포함하는 발현벡터 및 상기 발현벡터가 도입된 형질전환체를 제공한다.
본 발명에서 용어, "폴리뉴클레오티드"는 뉴클레오티드 단위체(monomer)가 공유결합에 의해 길게 사슬모양으로 이어진 뉴클레오티드의 중합체(polymer)로 일정한 길이 이상의 DNA(deoxyribonucleic acid) 또는 RNA(ribonucleic acid)가닥으로서, 상기 결합체를 코딩하는 폴리뉴클레오티드이다.
본 발명에서 제공하는 폴리뉴클레오티드는 특별히 이에 제한되지 않으나, 리파아제 TliA의 N-말단에 부포린 IIb, B0, 파라신 I, NKC 또는 NRC가 융합된 형태의 융합 폴리펩타이드를 암호화하는 폴리뉴클레오티드(서열번호 45 내지 49), 리파아제 M37의 N-말단에 부포린 IIb, B0, 파라신 I, NKC 또는 NRC가 융합된 형태의 융합 폴리펩타이드를 암호화하는 폴리뉴클레오티드(서열번호 50 내지 54), 리파아제 TliA의 C-말단에 부포린 IIb, B0, 파라신 I, NKC 또는 NRC가 융합된 형태의 융합 폴리펩타이드를 암호화하는 폴리뉴클레오티드(서열번호 55 내지 59), 리파아제 M37의 C-말단에 부포린 IIb, B0, 파라신 I, NKC 또는 NRC가 융합된 형태의 융합 폴리펩타이드를 암호화하는 폴리뉴클레오티드(서열번호 60 내지 64), 리파아제 TliA의 N-말단과 C-말단에 부포린 IIb, B0, 파라신 I, NKC 또는 NRC가 융합된 형태의 융합 폴리펩타이드를 암호화하는 폴리뉴클레오티드(서열번호 65 내지 69), 리파아제 M37의 N-말단과 C-말단에 부포린 IIb, B0, 파라신 I, NKC 또는 NRC가 융합된 형태의 융합 폴리펩타이드를 암호화하는 폴리뉴클레오티드(서열번호 70 내지 74)일 수 있다.
본 발명의 결합체를 코딩하는 폴리뉴클레오타이드는 코돈의 축퇴성(degeneracy)으로 인하여 또는 상기 결합체를 발현시키고자 하는 생물에서 선호되는 코돈을 고려하여, 코딩영역으로부터 발현되는 결합체의 아미노산 서열을 변화시키지 않는 범위 내에서 코딩영역에 다양한 변형이 이루어질 수 있고, 코딩영역을 제외한 부분에서도 유전자의 발현에 영향을 미치지 않는 범위 내에서 다양한 변형 또는 수식이 이루어질 수 있으며, 그러한 변형 유전자 역시 본 발명의 범위에 포함됨을 당업자는 잘 이해할 수 있을 것이다. 즉, 본 발명의 폴리뉴클레오티드는 이와 동등한 활성을 갖는 단백질을 코딩하는 한, 하나 이상의 핵산 염기가 치환, 결실, 삽입 또는 이들의 조합에 의해 변이될 수 있으며, 이들 또한 본 발명의 범위에 포함된다.
본 발명에서 제공하는 상기 결합체를 코딩하는 폴리뉴클레오티드를 포함하는 발현벡터는 특별히 이에 제한되지 않으나, 포유류 세포(예를 들어, 사람, 원숭이, 토끼, 래트, 햄스터, 마우스 세포 등), 식물 세포, 효모 세포, 곤충 세포 또는 박테리아 세포(예를 들어, 대장균 등)를 포함하는 진핵 또는 원핵세포에서 상기 폴리뉴클레오티드를 복제 및/또는 발현할 수 있는 벡터가 될 수 있고, 바람직하게는 숙주세포에서 상기 뉴클레오티드가 발현될 수 있도록 적절한 프로모터에 작동가능하도록 연결되며, 적어도 하나의 선별마커를 포함하는 벡터가 될 수 있다. 그 예로 파아지, 플라스미드, 코스미드, 미니-염색체, 바이러스 또는 레트로바이러스벡터 등에 상기 폴리뉴클레오티드가 도입된 형태가 될 수 있다.
본 발명에서 제공하는 상기 발현 벡터가 도입된 형질전환체는 특별히 이에 제한되지 않으나, 상기 발현벡터가 도입되어 형질전환된 대장균, 스트렙토미세스, 살모넬라 티피뮤리움 등의 박테리아 세포; 효모 세포; 피치아 파스토리스 등의 균류세포; 드로조필라, 스포도프테라 Sf9 세포 등의 곤충 세포; CHO(중국 햄스터 난소 세포, chinese hamster ovary cells), SP2/0(마우스 골수종), 인간 림프아구(human lymphoblastoid), COS, NSO(마우스 골수종), 293T, 보우 멜라노마 세포, HT-1080, BHK(베이비 햄스터 신장세포, baby hamster kidney cells), HEK(인간 배아신장 세포, human embryonic kidney cells), PERC.6(인간망막세포) 등의 동물 세포; 또는 식물 세포가 될 수 있다. 본 발명의 일 실시예에 따르면 대장균 세포를 숙주세포로 이용하였다(실시예 2).
본 발명에서 용어, "도입"은 상기 결합체를 코딩하는 폴리뉴클레오티드를 포함하는 벡터를 숙주세포에 전달하는 방법을 의미한다. 이와 같은 도입은 칼슘 포스페이트-DNA 공침전법, DEAE-덱스트란-매개 트랜스펙션법, 폴리브렌-매개 형질감염법, 전기충격법, 미세주사법, 리포좀 융합법, 리포펙타민 및 원형질체 융합법 등의 당 분야에 공지된 여러 방법에 의해 수행될 수 있다. 또한, 형질도입은 감염(infection)을 수단으로 하여 바이러스 입자를 사용하여 목적물을 세포 내로 전달시키는 것을 의미한다. 아울러, 유전자 밤바드먼트 등에 의해 벡터를 숙주세포 내로 도입할 수 있다. 본 발명에서 도입은 형질전환과 혼용되어 사용될 수 있다.
또 하나의 양태로서, 본 발명은 상기 양친매성 펩타이드-리파아제 결합체의 제조방법을 제공한다.
상기 제조방법은 바람직하게는 (a) 상기 형질전환체를 배양하는 단계; 및 (b) 상기 배양된 형질전환체 또는 이의 배양액으로부터 양친매성 펩타이드-리파아제 결합체를 회수하는 단계를 포함하나, 이에 제한되지 않는다.
상기 결합체를 회수하는 단계는 상기 배양물로부터 수득한 배양균체 또는 배양 상등액을 균체파쇄, 추출, 친화성크로마토그래피, 이온교환크로마토그래피, 겔여과 크로마토그래피, 소수성 크로마토그래피, 단백질 침전, 투석 등의 공지된 정제방법을 단독으로 또는 적당히 조합함으로써 수행될 수 있고, 회수된 목적 단백질의 확인은 SDS-PAGE, 웨스턴 블럿 등의 공지된 통상의 방법을 이용하여 수행될 수 있다.
본 발명의 일 실시예에서는 양친매성 펩타이드와 리파아제가 연결된 결합체인 파라신 I-TliA 리파아제, 부포린 IIb-TliA 리파아제, B0-TliA 리파아제, 파라신 I-M37 리파아제, 부포린 IIb-M37 리파아제, B0-M37 리파아제, NKC-M37 리파아제 및 NRC-M37 리파아제의 유전자를 각각 제작하고, 이를 포함하는 발현벡터(pET-Par-TliA, pET-Buf-TliA, pET-B0-TliA, pET-Par-M37, pET-Buf-M37, pET-B0-M37, pET-NKC-M37 및 pET-NRC-M37)를 각각 수득하였다. 상기 수득한 각 발현벡터를 대장균에 도입하여 각각의 형질전환체를 제작한 다음, 상기 제작된 각각의 형질전환체를 배양하고, 각각의 배양물로부터 각각의 융합 폴리펩타이드를 회수하여, 양친매성 펩타이드와 리파아제가 연결된 양친매성 펩타이드-리파아제 결합체를 제조하였다(실시예 1 및 2, 도 2 및 3).
또 하나의 양태로서, 본 발명은 상기 양친매성 펩타이드-리파아제 결합체와 지질을 반응시키는 단계를 포함하는, 상기 양친매성 펩타이드-리파아제 결합체를 이용한 지질의 분해방법을 제공한다.
상기 양친매성 펩타이드, 리파아제 및 양친매성 펩타이드-리파아제 결합체에 대해서는 상기에서 설명한 바와 같다. 상기 양친매성 펩타이드-리파아제 결합체는 양친매성 펩타이드로 인하여 지질 기질에 대한 향상된 접근성 및 반응성을 나타내므로(도 4 내지 8), 지질의 분해에 있어 효율적으로 사용될 수 있다. 또한, 양친매성 펩타이드가 결합되어 별도의 계면활성제의 사용이 요구되지 않으며, 리파아제 사용량이 적게 요구되므로, 경제적인 지질의 분해에 이용될 수 있다.
상기 지질은 본 발명의 리파아제에 분해될 수 있는 지질이라면 그 종류가 특별히 제한되지 않는다.
본 발명의 일 실시예에서는 상기 결합체의 리파아제 활성을 양친매성 펩타이드가 결합되지 않은 야생형 리파아제 활성과 비교하였다. 이때, 기질로는 파라니트로페닐 팔미테이트 및 올리브유를 사용하였다. 그 결과, 파라니트로페닐 팔미테이트를 기질로 사용할 경우, 리파아제 TliA를 포함하는 결합체는 야생형 리파아제 TliA에 비하여 리파아제 활성이 약 10% 정도 증가되었고(도 4), 리파아제 M37을 포함하는 결합체는 야생형 리파아제 M37에 비하여 리파아제 활성이 약 1.2 내지 4.2배 증가되는 결과를 보였다(도 6). 또한, 올리브유를 기질로 사용할 경우, 리파아제 TliA를 포함하는 결합체는 리파아제 TliA에 비하여 리파아제 활성이 약 35.5 내지 78배 증가되었고(도 5), 리파아제 M37을 포함하는 융합단백질은 리파아제 M37에 비하여 리파아제 활성이 약 2.2 내지 10.2배 증가되는 결과를 나타내었다(도 7). 상기와 같은 결과들은 본 발명의 결합체가 지질의 분해에 있어 효율적으로 사용될 수 있음을 뒷받침하는 결과이다.
또 하나의 양태로서, 본 발명은 상기 양친매성 펩타이드-리파아제 결합체를 이용한 바이오디젤의 생산방법을 제공한다.
상기 바이오디젤의 생산방법은 바람직하게는 상기 양친매성 펩타이드-리파아제 결합체와 유지 및 알코올을 반응시키는 단계를 포함하나, 이에 제한되지 않는다.
또한, 상기 바이오디젤의 생산방법은 (a) 상기 발현벡터를 숙주세포에 도입하여 형질전환된 형질전환체를 수득하는 단계; (b) 상기 수득한 형질전환체를 배양하여 배양물 또는 배양 상등액으로부터 리파아제를 회수하는 단계; 및 (c) 상기 회수된 리파아제를 사용하여 유지와 알코올로부터 바이오디젤을 생산하는 단계를 포함할 수 있으나, 이에 제한되지 않는다.
본 발명에서 바이오디젤의 생산에 사용되는 리파아제는 이에 제한되지 않으나, 자유로운 형태 또는 고정화된 형태로 사용할 수 있다. 또한, 상기 리파아제의 고정화는 당업계에서 통상적으로 사용하는 다양한 방법을 사용할 수 있으며, 이에 제한되지 않는 다양한 응용이 가능하다. 예를 들어, 흡착법, 포괄법 등의 물질적인 방법 및 공유결합법, 가교연결방법 등의 화학적인 방법을 사용할 수 있다.
상기 유지는 이에 제한되지는 않으나, 바람직하게는 천연유지, 가공유지 및폐유지 중에서 선택된 것을 사용할 수 있고, 보다 바람직하게는 대두유, 채종유, 팜유 등을 사용할 수 있다. 또한, 상기 알코올은 이에 제한되지는 않으나, 바람직하게는 탄소 수가 2개 내지 8개인 알코올을 사용할 수 있고, 보다 바람직하게는 탄소 수가 2개 내지 4개인 알코올을 사용할 수 있으며, 구체적으로 에탄올, 메탄올, 1-프로판올, 이소-프로판올, 1-부탄올, 2-부탄올, 이소-부탄올 또는 테르-부탄올 등을 사용할 수 있다. 상기 알코올은 당업계에 공지된 다양한 공급 방식을 이용하여 공급할 수 있으며, 여러 단계에 걸쳐서 공급하거나 연속적으로 공급하는 방식을 이용할 수 있다.
본 발명의 일 실시예에서는 유지로서, 올리브오일 또는 오일 폐기물을 사용하였으며, 알코올로서 메탄올을 사용하였다(실시예 5). 또한, 높은 농도의 메탄올 배지에서 불안정한 일반적인 리파아제와 달리 본 발명의 리파아제는 CalB 리파아제에 비하여 3.3%, 5% 및 10%에서도 높은 활성을 유지하였다(도 9). 또한, 3 단계 메탄올 공급 방법(1 당량)을 사용하여 본 발명의 대표적인 결합체인 NKC-M37 리파아제의 올리브오일의 바이오디젤로의 생산 효율을 확인한 결과, 야생형 M37 리파아제에 비하여 21시간이나 빨리 95%의 전환 수율에 도달하였다(도 10 a). 또한, 3 단계 메탄올 공급 방법에 비하여 보다 경제적인 2 단계 메탄올 공급 방법을 사용하였을 때, 야생형 M37 리파아제에 비하여 27시간 빨리 최대 전환 수율에 도달하는 결과를 보였으며(도 10 b), 1 단계 메탄올 공급 방법을 사용하였을 때도 야생형 M37 리파아제 비하여 훨씬 빠르게 최대 전환 수율에 도달하는 결과를 나타내었다(도 10 c). 또한, 공급 원료가 훨씬 저렴하나 불순물, 유리 지방산 및 함수량 등을 포함하여 리파아제의 활성에 영향을 미쳐 바이오디젤의 생산에 사용이 어려운 것으로 알려진 오일 폐기물을 기질로서 사용하였다. 그 결과, 오일 폐기물에 본 발명의 대표적인 양친매성 펩타이드-리파아제 결합체인 NKC-M37 리파아제를 사용하였을 때, 올리브유 및 팜유와 비슷한 정도의 바이오디젤이 생산되는 결과를 나타내어, 본 발명의 결합체가 바이오디젤 생산에 있어 경제적 가치를 지니는 결합체임을 확인하였다(실시예 5).
본 발명의 융합 폴리펩타이드는 종래의 리파아제에 기질의 반응성을 증가시킬 수 있는 양친매성 펩타이드가 융합된 형태로서, 리파아제의 효소활성을 현저하게 증가시키므로 바이오디젤 생산과 같은 리파아제가 필요한 분야에 있어 소량을 사용하여도 충분한 지질 전환 수율을 보일 수 있을 뿐만 아니라, 리파아제의 활성을 증가시키기 위한 별도의 계면활성제를 사용하지 않을 수 있으므로, 리파아제의 경제적인 사용에 널리 활용될 수 있을 것이다.
도 1은, 양친매성 펩타이드(amphipathic peptide)을 Schiffer-Edmundson wheel projection으로 나타낸 도로서, 1번은 펩타이드의 1번 잔기를 나타내며, 검은 동그라미(filled circle)은 소수성 아미노산을, 흰 동그라미는 친수성 아미노산을 나타낸다.
도 2는, 활성이 증대된 리파아제를 발현하기 위한 벡터의 유전자 구조도이다.
도 3은, 본 발명에서 발현된 펩타이드-리파아제 TliA 발현양상을 나타내는 SDS-PAGE 사진이다.
도 4는, 본 발명에 따라 제조된 TliA 리파아제의 지질분해활성을, 리파아제 반응시 발색하는 파라니트로페닐 팔미테이트(para-Nitrophenyl palmitate)를 이용하여 측정한 결과를 나타내는 그래프이다.
도 5는, 본 발명에 따라 제조된 TliA 리파아제의 지질분해활성 증대를 pH-STAT으로 측정한 결과를 나타내는 그래프이다.
도 6은, 본 발명에 따라 제조된 M37 리파아제의 지질분해활성을, 리파아제 반응 시 발색하는 파라니트로페닐 팔미테이트를 이용하여 측정한 결과를 나타내는 그래프이다.
도 7은 본 발명에 따라 제조된 M37 리파아제의 지질분해활성 증대를 pH-STAT으로 측정한 결과를 나타내는 그래프이다.
도 8은, NKC-M37 리파아제의 지질 입자에 대한 위치를 형광 현미경으로 관찰한 결과를 나타내는 도이다.
도 9는, 칸디다 안탁티카(Candida antarctica) 유래의 CalB 리파아제(a), 야생형 M37 리파아제(b) 및 NKC-M37 리파아제(c)의 효소 활성을 나타낸 도로서, 40℃에서 12시간 내의 시간에서 반응시킨 결과를 나타낸 것이다. CalB 리파아제, 야생형 M37 리파아제 및 NKC-M37 리파아제의 가수분해활성을 0%, 3.3%, 5% 및 10%의 메탄올-수용액에서 측정하였다.
도 10은, 야생형 M37 리파아제 및 NKC-M37 리파아제를 이용하여 올리브 오일을 사용하여 바이오디젤 생산을 확인한 도이다. 바이오디젤의 양은 가스 크로마토그래피 및 박층 크로마토그래피를 이용하여 정량적으로 분석하였다. 도 10 a는 3 단계 에스테르교환반응을 시간 의존적으로 수행한 결과를 나타낸 것으로, 야생형 M37 리파아제는 흰 동그라미(open circle)로, NKC-M37 리파아제는 검은색 동그라미(closed circle)로 나타내었다. 또한, 야생형 M37 리파아제(왼쪽) 및 NKC-M37 리파아제(오른쪽)을 사용한 에스테르 교환반응에 대한 반응 혼합물의 TLC 분석을 나타내었다. 도 10 b는 야생형 M37 리파아제 및 NKC-M37 리파아제를 사용하여 올리브 오일의 2 단계 에스테르 교환반응을 시간 의존적으로 수행한 결과를 나타내는 것이다. 도 10 c는 야생형 M37 리파아제 및 NKC-M37 리파아제를 사용하여 올리브 오일의 1 단계 에스테르 교환반응을 시간 의존적으로 수행한 결과를 나타내는 것이다.
도 2는, 활성이 증대된 리파아제를 발현하기 위한 벡터의 유전자 구조도이다.
도 3은, 본 발명에서 발현된 펩타이드-리파아제 TliA 발현양상을 나타내는 SDS-PAGE 사진이다.
도 4는, 본 발명에 따라 제조된 TliA 리파아제의 지질분해활성을, 리파아제 반응시 발색하는 파라니트로페닐 팔미테이트(para-Nitrophenyl palmitate)를 이용하여 측정한 결과를 나타내는 그래프이다.
도 5는, 본 발명에 따라 제조된 TliA 리파아제의 지질분해활성 증대를 pH-STAT으로 측정한 결과를 나타내는 그래프이다.
도 6은, 본 발명에 따라 제조된 M37 리파아제의 지질분해활성을, 리파아제 반응 시 발색하는 파라니트로페닐 팔미테이트를 이용하여 측정한 결과를 나타내는 그래프이다.
도 7은 본 발명에 따라 제조된 M37 리파아제의 지질분해활성 증대를 pH-STAT으로 측정한 결과를 나타내는 그래프이다.
도 8은, NKC-M37 리파아제의 지질 입자에 대한 위치를 형광 현미경으로 관찰한 결과를 나타내는 도이다.
도 9는, 칸디다 안탁티카(Candida antarctica) 유래의 CalB 리파아제(a), 야생형 M37 리파아제(b) 및 NKC-M37 리파아제(c)의 효소 활성을 나타낸 도로서, 40℃에서 12시간 내의 시간에서 반응시킨 결과를 나타낸 것이다. CalB 리파아제, 야생형 M37 리파아제 및 NKC-M37 리파아제의 가수분해활성을 0%, 3.3%, 5% 및 10%의 메탄올-수용액에서 측정하였다.
도 10은, 야생형 M37 리파아제 및 NKC-M37 리파아제를 이용하여 올리브 오일을 사용하여 바이오디젤 생산을 확인한 도이다. 바이오디젤의 양은 가스 크로마토그래피 및 박층 크로마토그래피를 이용하여 정량적으로 분석하였다. 도 10 a는 3 단계 에스테르교환반응을 시간 의존적으로 수행한 결과를 나타낸 것으로, 야생형 M37 리파아제는 흰 동그라미(open circle)로, NKC-M37 리파아제는 검은색 동그라미(closed circle)로 나타내었다. 또한, 야생형 M37 리파아제(왼쪽) 및 NKC-M37 리파아제(오른쪽)을 사용한 에스테르 교환반응에 대한 반응 혼합물의 TLC 분석을 나타내었다. 도 10 b는 야생형 M37 리파아제 및 NKC-M37 리파아제를 사용하여 올리브 오일의 2 단계 에스테르 교환반응을 시간 의존적으로 수행한 결과를 나타내는 것이다. 도 10 c는 야생형 M37 리파아제 및 NKC-M37 리파아제를 사용하여 올리브 오일의 1 단계 에스테르 교환반응을 시간 의존적으로 수행한 결과를 나타내는 것이다.
이하, 본 발명을 하기 실시예에서 보다 구체적으로 설명한다. 그러나 이들 실시예는 본 발명의 이해를 돕기 위한 것일 뿐, 이들에 의해 본 발명이 한정되는 것은 아니다.
실시예 1: 다양한 양친매성 펩타이드 유도체와 리파아제가 융합된 리파아제 단백질의 개발
실시예
1-1:
펩타이드
유도체 제작
여러 항균 펩타이드들은 강력한 항균활성을 가지고 있으면서 동시에 소수기와 친수기를 가지는 양친매성의 α-helix 구조를 이루고 있다. 본 발명자들은 그 중 특히 우수한 양친매성을 나타내는, 서열번호 1의 아미노산 서열로 기재된 부포린 IIb(대한민국 등록특허 제10-314721호), 상기 부포린 IIb를 변형시킨 서열번호 2의 아미노산 서열로 기재된 펩타이드 B0, 서열번호 3의 아미노산 서열로 기재된 파라신 I(대한민국 등록특허 제10-330136호), 서열번호 4의 아미노산 서열로 기재된 NKC 및 서열번호 5의 아미노산 서열로 기재된 NRC(대한민국 등록특허 제10-836596호)를 제작하였고(표 1), 이를 이용하여 지질분해 활성이 증대된 융합 폴리펩타이드를 제작하였다.
본 발명에서 사용된 양친매성 펩타이드 | ||
명칭 | 아미노산 서열(N-C) | 서열번호 |
부포린 IIb | RAGLQFPVGRLLRRLLRRLLR | 1 |
B0 | RAGLQFPVG | 2 |
파라신 I | KGRGKQGGKVRAKAKTRSS | 3 |
NKC | APKAMKLLKKLLKLQKKGI | 4 |
NRC | APKAMRLLRRLLRLQKKGI | 5 |
상기 5가지 양친매성 펩타이드의 α-helix 구조는 Schiffer-Edmundson wheel projection을 통하여 시각화할 수 있으며, 이를 도 1에 나타내었다. 도 1에 나타낸 Wheel에서 연이은 아미노산(consecutive amino acids)은 싸이클(cycle) 당 3.6 아미노산을 가지도록, 서로 100°가량 떨어져 있다.
실시예
1-2:
펩타이드와
리파아제가 융합된 유전자의 제작
효소의 활성이 강하며 바이오디젤 생산 시 전환효율이 뛰어난 슈도모나스 플루오레슨스(Pseudomonas fluorescens) 유래의 리파아제 TliA와 메탄올 내성이 강한 포토박테리움 리포리티쿰(Photobacterium lipolyticum) 유래의 리파아제 M37의 유전자를 포함하고 있는 플라스미드 pHOPE(Eom, GT, et al., Applied Environ. Microbiol., 71:3468-3474, 2005)와 pEML37(Yang KS, et al., J. Biosci. Bioeng., 107:599-604, 2009)를 주형으로 이용하여 재조합 PCR(Polymerase Chain Reaction)을 통해 상기 리파아제 유전자와 선별한 펩타이드 유전자를 융합시켰다. 구체적인 실험 방법은 하기와 같으며, 본 실험에서 사용한 프라이머를 하기 표 2에 나타내었다.
프라이머 | 서열(5'->3') | 서열번호 |
pET16-L | 5'-cgtagaggatcgagatctcgatcc-3' | 서열번호 75 |
pET-R(Par) | 5'-gccttcgcacgcaccttgcctccctgtttgcctcttcctttacgaccttcgatatggccg-3' | 서열번호 76 |
TliA-rev(Par) | 5'-gggaggcaaggtgcgtgcgaaggcaaagacacgttcatccggtgtatttgactacaagaacc-3' | 서열번호 77 |
NdeI-TliA-for | 5'-cttaaggcatatgtcaactgatcagcacaccctcg-3' | 서열번호 78 |
pET-R(Buf) | 5'-cgacgcagcagacgaccaaccgggaactgcagaccagcacgacgaccttcgatatggccg-3' | 서열번호 79 |
TliA-rev(Buf) | 5'-cccggttggtcgtctgctgcgtcgtctgctgcgtcgtctgctgcgtggtgtatttgactacaagaacc-3' | 서열번호 80 |
pET-R(B0) | 5'-caaccgggaactgcagaccagcacgacgaccttcgatatggccg-3' | 서열번호 81 |
TliA-rev(B0) | 5'-gctggtctgcagttcccggttggtggtgtatttgactacaagaacc-3' | 서열번호 82 |
pET-R(con) | 5'-acgaccttcgatatggccg-3' | 서열번호 83 |
TliA-rev(con) | 5'-cggccatatcgaaggtcgtggtgtatttgactacaagaacc-3' | 서열번호 84 |
M37-rev(Buf) | 5'-cccggttggtcgtctgctgcgtcgtctgctgcgtcgtctgctgcgtgcatctccacgcgccaatgatg-3' | 서열번호 85 |
Nde-M37-for | 5'-cttaaggcatatgttataacaaacccgcgatcgca-3' | 서열번호 86 |
M37-rev(Par) | 5'-gggaggcaaggtgcgtgcgaaggcaaagacacgttcatccgcatctccacgcgccaatgatg-3' | 서열번호 87 |
M37-rev(B0) | 5'-gctggtctgcagttcccggttggtgcatctccacgcgccaatgatg-3' | 서열번호 88 |
M37-rev(NKC) | 5'-actgttgaagaaattgctgaaattacagaaaaaaggcattgcatctccacgcgccaatgatg-3' | 서열번호 89 |
M37-rev(NRC) | 5'-tctgttgcgtcgcttgctgcgtttacagaaaaaaggcattgcatctccacgcgccaatgatg-3' | 서열번호 90 |
M37-rev(con) | 5'-gcagcggccatatcgaaggtcgtgcatctccacgcgccaatg-3' | 서열번호 91 |
실시예
1-2-1:
펩타이드
파라신 I과 리파아제
TliA가
융합된 유전자의 제작
파라신 I과 리파아제 유전자 TliA가 융합된 유전자를 만들기 위하여 pET16b 벡터(Merck biosciences)를 주형으로 하고, DNA 프라이머 pET16-L과 pET-R(Par)를 이용하여 T7 프로모터-리보좀 결합부위-His tag와 연결된 펩타이드 유전자 단편을 PCR 증폭하였다.
또한, pHOPE 플라스미드를 주형으로 하고, 프라이머 TliA-rev(Par)와 Nde-TliA-for를 이용하여, 펩타이드 유전자 일부와 리파아제 유전자가 결합된 DNA 단편을 PCR을 이용하여 합성하였다.
상기 증폭된 두 개의 DNA 단편을 주형으로 하여 프라이머 pET16-L과 Nde-TliA-for를 이용해 T7 프로모터-리보좀 결합부위-His tag-파라신 I 펩타이드-리파아제 유전자가 결합된 DNA 단편을 재조합 PCR 증폭하였다.
실시예 1-2-2: 펩타이드 부포린 IIb와 리파아제 TliA가 융합된 유전자의 제작
부포린 IIb와 리파아제 유전자 TliA가 융합된 유전자를 만들기 위하여 pET16b 벡터를 주형으로 하고, DNA 프라이머 pET16-L과 pET-R(Buf)를 이용하여 T7 프로모터-리보좀 결합부위-His tag과 연결된 펩타이드 유전자 단편을 PCR 증폭하였다.
또한, pHOPE 플라스미드를 주형으로 하고, 프라이머 TliA-rev(Buf)와 Nde-TliA-for를 이용하여 펩타이드 유전자 일부와 리파아제 유전자가 결합된 DNA 단편을 PCR을 이용하여 합성하였다.
상기 증폭된 두 개의 DNA 단편을 주형으로 하여 프라이머 pET16-L과 Nde-TliA-for를 이용해 T7 프로모터-리보좀 결합부위-His tag-부포린 IIb 펩타이드-리파아제 유전자가 결합된 DNA 단편을 재조합 PCR 증폭하였다.
실시예
1-2-3:
펩타이드
B0와 리파아제
TliA가
융합된 유전자의 제작
B0와 리파아제 유전자 TliA가 융합된 유전자를 만들기 위하여 pET16b 벡터를 주형으로 하고, DNA 프라이머 pET16-L과 pET-R(B0)를 이용하여 T7 프로모터-리보좀 결합부위-His tag과 연결된 펩타이드 유전자 단편을 PCR 증폭하였다.
또한, pHOPE 플라스미드를 주형으로 하고, 프라이머 TliA-rev(B0)와 Nde-TliA-for를 이용하여 펩타이드 유전자 일부와 리파아제 유전자가 결합된 DNA 단편을 PCR을 이용하여 합성하였다.
상기 증폭된 두 개의 DNA 단편을 주형으로 하여 프라이머 pET16-L과 Nde-TliA-for를 이용해 T7 프로모터-리보좀 결합부위-His tag-B0 펩타이드-리파아제 유전자가 결합된 DNA 단편을 재조합 PCR 증폭하였다.
실시예
1-2-4: 리파아제
TliA를
포함하는 대조군 유전자의 제작
펩타이드가 붙지 않은 대조군(T7 프로모터-리보좀 결합부위-His tag-리파아제 TliA) DNA를 증폭시키기 위하여, pET16b 벡터를 주형으로 하고, DNA 프라이머 pET16-L과 pET-R(con)를 이용하여 T7 프로모터-리보좀 결합부위-His tag이 연결된 유전자 단편을 PCR 증폭하였다.
또한, pHOPE 플라스미드를 주형으로 하고, 프라이머 TliA-rev(con)와 Nde-TliA-for를 이용하여 리파아제 유전자를 포함하는 DNA 단편을 PCR을 이용하여 합성하였다.
상기 증폭된 두 개의 DNA 단편을 주형으로 하여 프라이머 pET16-L과 Nde-TliA-for를 이용해 T7 프로모터-리보좀 결합부위-His tag-리파아제 유전자가 결합된 DNA 단편을 재조합 PCR 증폭하였다.
실시예
1-2-5:
펩타이드
부포린
IIb와
리파아제 M37이 융합된 유전자의 제작
부포린 IIb와 M37 리파아제가 융합된 유전자를 만들기 위하여, pET16b 벡터를 주형으로 DNA 프라이머 pET16-L과 pET-R(Buf)를 이용하여 T7 프로모터-리보좀 결합부위-His tag과 연결된 펩타이드 유전자 단편을, pEML37 플라스미드를 주형으로 프라이머 M37-rev(Buf)와 Nde-M37-for를 이용하여 펩타이드 유전자 일부와 리파아제 유전자가 결합된 DNA 단편을 각각 PCR 증폭한 뒤, 증폭한 두 개의 DNA 단편을 주형으로 하여 프라이머 pET16-L과 Nde-M37-for를 이용해 T7 프로모터-리보좀 결합부위-His tag-부포린 IIb-리파아제 유전자가 결합된 DNA 단편을 재조합 PCR 증폭하였다.
실시예
1-2-6:
펩타이드
파라신 I과 리파아제 M37이 융합된 유전자의 제작
파라신 I과 M37 리파아제가 융합된 유전자를 만들기 위하여, pET16b 벡터를 주형으로 DNA 프라이머 pET16-L과 pET-R(Par)를 이용하여 T7 프로모터-리보좀 결합부위-His tag과 연결된 펩타이드 유전자 단편을, pEML37 플라스미드를 주형으로 프라이머 M37-rev(Par)와 Nde-M37-for를 이용하여 펩타이드 유전자 일부와 리파아제 유전자가 결합된 DNA 단편을 각각 PCR 증폭한 뒤, 증폭한 두 개의 DNA 단편을 주형으로 하여 프라이머 pET16-L과 Nde-M37-for를 이용해 T7 프로모터-리보좀 결합부위-His tag-파라신 I-리파아제 유전자가 결합된 DNA 단편을 재조합 PCR 증폭하였다.
실시예
1-2-7:
펩타이드
B0와 리파아제 M37이 융합된 유전자의 제작
펩타이드 B0와 M37 리파아제가 융합된 유전자를 만들기 위하여, pET16b 벡터를 주형으로 DNA 프라이머 pET16-L과 pET-R(B0)를 이용하여 T7 프로모터-리보좀 결합부위-His tag과 연결된 펩타이드 유전자 단편을, pEML37 플라스미드를 주형으로 프라이머 M37-rev(B0)와 Nde-M37-for를 이용하여 펩타이드 유전자 일부와 리파아제 유전자가 결합된 DNA 단편을 각각 PCR 증폭한 뒤, 증폭한 두 개의 DNA 단편을 주형으로 하여 프라이머 pET16-L과 Nde-M37-for를 이용해 T7 프로모터-리보좀 결합부위-His tag-B0-리파아제 유전자가 결합된 DNA 단편을 재조합 PCR 증폭하였다.
실시예
1-2-8:
펩타이드
NKC와
리파아제 M37이 융합된 유전자의 제작
NKC와 M37 리파아제가 융합된 유전자를 만들기 위하여, pET16b 벡터를 주형으로 DNA 프라이머 pET16-L과 pET-R(NKC)를 이용하여 T7 프로모터-리보좀 결합부위-His tag과 연결된 펩타이드 유전자 단편을, pEML37 플라스미드를 주형으로 프라이머 M37-rev(NKC)와 Nde-M37-for를 이용하여 펩타이드 유전자 일부와 리파아제 유전자가 결합된 DNA 단편을 각각 PCR 증폭한 뒤, 증폭한 두 개의 DNA 단편을 주형으로 하여 프라이머 pET16-L과 Nde-M37-for를 이용해 T7 프로모터-리보좀 결합부위-His tag-NKC-리파아제 유전자가 결합된 DNA 단편을 재조합 PCR 증폭하였다.
실시예
1-2-9:
펩타이드
NRC와 리파아제 M37이 융합된 유전자의 제작
NRC와 M37 리파아제가 융합된 유전자를 만들기 위하여, pET16b 벡터를 주형으로 DNA 프라이머 pET16-L과 pET-R(NRC)를 이용하여 T7 프로모터-리보좀 결합부위-His tag과 연결된 펩타이드 유전자 단편을, pEML37 플라스미드를 주형으로 프라이머 M37-rev(NRC)와 Nde-M37-for를 이용하여 펩타이드 유전자 일부와 리파아제 유전자가 결합된 DNA 단편을 각각 PCR 증폭한 뒤, 증폭한 두 개의 DNA 단편을 주형으로 하여 프라이머 pET16-L과 Nde-M37-for를 이용해 T7 프로모터-리보좀 결합부위-His tag-NRC-리파아제 유전자가 결합된 DNA 단편을 재조합 PCR 증폭하였다.
실시예
1-2-10: 리파아제 M37을 포함하는 대조군 유전자의 제작
펩타이드가 붙지 않은 대조군(T7 프로모터-리보좀 결합부위-His tag-리파아제 M37) DNA를 증폭시키기 위하여, pET16b 벡터를 주형으로 DNA 프라이머 pET16-L과 pET-R(NRC)를 이용하여 T7 프로모터-리보좀 결합부위-His tag과 연결된 펩타이드 유전자 단편을 증폭하고, pEML37 플라스미드를 주형으로 프라이머 M37-rev(con)와 Nde-M37-for를 이용하여 리파아제 유전자를 PCR 증폭한 뒤, 증폭한 두 개의 DNA 단편을 주형으로 하여 프라이머 pET16-L과 Nde-M37-for를 이용해 T7 프로모터-리보좀 결합부위-His tag- M37 리파아제 유전자가 결합된 DNA 단편을 PCR 증폭하였다.
실시예
1-3:
펩타이드와
리파아제 융합 단백질의 발현 플라스미드 제작
상기 실시예 1-2에서 제작된 각 T7 프로모터-리보좀 결합부위-His tag-펩타이드-리파아제 결합 DNA를 제한효소 BglII와 NdeI을 이용하여 자르고, BglII와 NdeI을 이용하여 잘린 pET16b 벡터에 클로닝하였다(도 2). 도 2는 활성이 증대된 리파아제를 발현하기 위한 벡터의 유전자 구조도로서, 리파아제 유전자의 아미노말단(N-terminus)에 조건발현 프로모터(conditional promoter) 하위에 양친매성 펩타이드 유전자를 융합시키고 발현된 단백질의 순수분리를 위해서 펩타이드의 앞쪽에 히스티딘 태그(histidine-tag)를 융합시키고 발현 및 순수분리 이후 히스티딘 태그를 제거하기 위해 히스티딘 태그와 펩타이드 유전자 사이에 단백질 분해효소 Factor Xa 인식부위를 삽입하여, 니켈 칼럼(Ni-column)에 결합하여 분리한 히스티딘 태그-펩타이드-리파아제 구조의 단백질에 Factor Xa 효소를 처리하여 양친매성 펩타이드-리파아제 구조의 원하는 단백질을 순수 분리 하도록 하였다. 그리고 이러한 유전자를 조건발현용 프로모터 하위에 둬, 우리가 원하는 조건에서 양친매성 펩타이드와 융합되어 활성이 증대된 리파아제가 발현되도록 하였다.
상기 클로닝된 플라스미드를 대장균 XL1-Blue(invitrogen 사에서 구매)에 도입하여 형질전환시키고, 항생제 암피실린을 포함하는 LB 평판배지에서 선별하였다. 정확한 플라스미드 제작을 확인하기 위하여 평판배지 상에서 자라는 콜로니에서 플라스미드를 분리한 다음 제한효소 BglII와 NdeI을 이용하여 DNA 단편의 크기를 확인하고 DNA 염기서열 분석을 통하여 정확하게 검증하였다.
이에 따라, 각각의 TliA 리파아제(대조군), 파라신 I-TliA 리파아제, 부포린 IIb-TliA 리파아제, B0-TliA 리파아제, M37 리파아제(대조군), 파라신 I-M37 리파아제, 부포린 IIb-M37 리파아제, B0-M37 리파아제, NKC-M37 리파아제, NRC-M37 리파아제를 발현하기 위하여 완성된 플라스미드를 각각 pET-TliA, pET-Par-TliA, pET-Buf-TliA, pET-B0-TliA, pET-M37, pET-Par-M37, pET-Buf-M37, pET-B0-M37, pET-NKC-M37 및 pET-NRC-M37로 명명하였다.
실시예
2:
펩타이드
-리파아제 융합 단백질의 분리 및 효소 활성 분석
실시예
2-1:
펩타이드
-리파아제 융합 단백질의 발현
펩타이드-리파아제 융합 단백질을 발현하기 위해 앞서 제조한 플라스미드를 전기천공법(electroporation)을 통해 E. coli BL21(DE3)(Merck biosciences)에 도입하고, 도말하여 단일 콜로니를 선별하였다. 37℃ LB 액체 배지에 접종한 후, 600nm의 파장에서 흡광도가 0.4일 때까지 키우고, 1mM IPTG(isopropyl-1-thio-β-D-galactopyranoside)를 넣어 펩타이드가 결합된 리파아제의 발현을 유도하였다. 4시간 후 대장균을 수확한 다음, 10% SDS-PAGE로 발현 여부를 확인하였다. 그 다음, Bradford 분석법을 이용하여 단백질의 농도를 측정하였다.
그 결과, 펩타이드-리파아제 융합 단백질은 대장균에서 봉입체(inclusion body)의 형태로 발현되는 것을 확인하였다(도 3). 도 3은 본 발명에서 발현된 펩타이드-리파아제 TliA 융합단백질 발현양상을 나타내는 SDS-PAGE 사진으로서, M번 레인은 단백질 크기 마커(size marker)를 나타내고, 1 내지 4번 레인은 각각 IPTG로 유도되지 않은 세포의 파쇄액, IPTG로 유도된 세포의 파쇄액, IPTG로 유도된 세포의 용해 상층액 부분 및 IPTG로 유도된 세포의 봉입체 부분을 나타낸다. 여기서 Par은 파라신 I을, Buf는 부포린 IIb를 나타내는 것이다.
실시예
2-2:
펩타이드
-리파아제 융합 단백질의 분리
재조합 단백질을 분리하기 위하여 배양한 대장균을 용해 완충용액(50 mM NaH2PO4, 300mM NaCl, pH 8.0)에 현탁시키고 초음파로 분쇄하였다(520초, 0.5싸이클, 50% amplitude). 초음파로 분쇄한 파쇄액을 4℃, 12,000rpm에서 20분간 원심분리하여 용해 상층액(soluble supernatant)과 봉입체(inclusion body)를 포함하는 비용해 부분으로 나눴다. 비용해 부분에 존재하는 펩타이드-리파아제 융합 단백질을 세정완충용액(100mM NaH2PO4, 10mM Tris-Cl, 8M urea, pH 6.3)에 녹이고, 원심분리하여 세포 찌꺼기를 제거했다. 원심분리 후, NTA(Ni2 +nitriloacetic acid) 아가로스 컬럼(Qiagen, Hilden, Germany)을 이용하여 상층액 속의 펩타이드-리파아제 융합 단백질을 분리하고, 세포막 투석(dialysis)을 통하여 남아있는 요소를 제거했다. 융합 단백질 분리에 사용된 His Tag을 제거하기 위해서 Factor Xa 효소(New England Biolab, USA)를 이용하였고, 순수 분리된 단백질을 10% SDS-PAGE에서 확인하고 N-말단 아미노산 염기서열을 분석하여 확인하였다.
실시예
2-3:
펩타이드
-리파아제 융합 단백질의 리파아제 활성 측정
실시예
2-3-1:
파라니트로페닐
팔미테이트를
이용한 리파아제의 활성 측정
리파아제의 활성은 일차적으로 파라니트로페닐 팔미테이트(p-nitrophenyl palmitate)의 발색반응을 이용하여 측정하였다. 10mM의 농도로 아세토나이트릴(acetonitrile)에 녹인 파라니트로페닐 팔미테이트 용액과 에탄올, 50mM Tris-HCl 용액(pH 8.5)을 1:4:95의 비율로 섞어 사용하였다. 파라니트로페닐 팔미테이트를 포함하는 용액 0.8㎖에 0.2㎖의 융합 단백질 용액을 혼합한 뒤, 45℃에서 10분간 반응하고 405nm에서 흡광도를 측정한 다음, 그 결과를 도 4 및 도 6에 각각 나타내었다. 도 4는, 본 발명의 양친매성 펩타이드를 융합시킨 TliA 리파아제의 지질분해활성을 리파아제 반응 시 발색하는 파라니트로페닐 팔미테이트를 이용하여 측정한 결과를 나타내는 그래프이고, 도 6은 본 발명의 양친매성 펩타이드를 융합시킨 M37 리파아제의 지질분해활성을 리파아제 반응시 발색하는 파라니트로페닐 팔미테이트를 이용하여 측정한 결과를 나타내는 그래프이다.
그 결과, 파라신 I이 융합된 파라신-TliA 융합 리파아제(Par-lip)는 상온에서도 반응 최적온도인 45℃에서와 비슷한 활성을 나타냄을 확인하였으며, 파라신 I과 B0가 각각 융합된 TliA의 경우 대조군에 비하여, 45℃에서 활성이 약 10% 증가함을 확인하였다(도 4). 또한, 도 6에서 보듯이, B0가 융합된 M37 리파아제(B0-M37), 부포린 IIb가 융합된 M37 리파아제(Buf-M37), NKC가 융합된 M37 리파아제(NKC-M37), NRC가 융합된 M37 리파아제(NRC-M37) 및 파라신 I이 융합된 M37 리파아제(Par-M37) 각각은 대조군에 비하여 4.2배, 2.7배, 6.3배, 1.3배 및 1.2배 증가함을 나타내었다. 상기와 같은 결과들은, 양친매성 펩타이드를 융합시킨 본 발명의 리파아제는 양친매성 펩타이드를 융합시키지 않은 리파아제에 비하여 뛰어난 리파아제 활성을 나타낼 수 있음을 뒷받침하는 것이다.
실시예
2-3-2: 올리브유 가수분해를 이용한 리파아제의 활성 측정
리파아제의 활성을 보다 정량적으로 측정하기 위해서 리파아제에 의한 올리브유의 가수분해 후 유리되는 지방산의 양을 측정하였다. 올리브유 에멀션(emulsion)은 20mM NaCl, 1mM CaCl2, 0.5%(w/v) 아라비아 고무(gum arabic) 용액 450㎖에 5㎖의 올리브유를 넣고 워어링 혼합기(Waring blender)에서 최고 속도로 2분간 유탁시켜 준비하고, 10mM NaOH 용액을 이용하여 pH를 8.0이 되도록 맞추고, 상기 올리브유 에멀션(emulsion)에 리파아제를 넣어 유리되는 지방산의 양을 pH 적정기(718 Stat Titrino, Metrohm)를 이용하여 50℃에서 5분간 측정하였고 그 결과를 각각 도 5 및 도 7에 나타내었다. 도 5는 본 발명에 따라 제조된 TliA 리파아제의 지질분해활성 증대를 pH-STAT으로 측정한 결과를 나타내는 그래프이고, 도 7은 본 발명에 따라 제조된 M37 리파아제의 지질분해활성 증대를 pH-STAT으로 측정한 결과를 나타내는 그래프이다.
그 결과, 도 5에 나타낸 바와 같이, B0-TliA, Par-TliA, Buf-TliA 융합 단백질의 경우, TliA에 비해서 그 활성이 각각 78±11.3배, 68.5±2.1배, 35.5±19.1배 증가함을 알 수 있었다. 또한, 도 7에서 보듯이, B0-M37, Buf-M37, NKC-M37, NRC-M37, Par-M37 융합 단백질의 경우 M37에 비해 그 활성이 각각 2.8배, 4.2배, 10.2배, 4.1배, 2.2배 증가함을 알 수 있었다.
이러한 결과는 양친매성 펩타이드를 융합시킨 본 발명의 리파아제는 양친매성 펩타이드를 융합시키지 않은 리파아제에 비해 뛰어난 지질 분해 활성을 나타낼 수 있음을 뒷받침하는 것이다.
실시예 3: 펩타이드 NKC가 융합된 M37 리파아제( NKC -M37)의 지질 기질(lipid substrates)에 대한 접근성 증대 확인
M37 리파아제의 활성 부위(active site)는 리드 나선(Lid helix, α3)에 가려져 있으며, M37 리파아제의 리드 부위 주변에 위치한 대부분의 소수성 잔기(Ile97, Trp100, Leu101, and Phe102)는 숨겨져 있으며, 활성 부위 주변에 위치해있다. 이러한 사실들은 기질이 리파아제에 결합하게 되면 상기와 같은 소수성 잔기들이 노출되어 넓은 소수성 표면(wide hydrophobic surface)을 형성할 것이라는 것을 시사하는 것으로 소수성 기질들이 M37 리파아제의 활성 부위로 접근하기 위해서는 어느정도의 구조 변화(conformational change)가 필요하다. 본 발명에서는 양친매성 펩타이드인 NKC를 M37 리파아제에 접합(conjugation)시켰으므로, 소수성 기질들이 상기 리파아제의 활성 부위로 접근하기 용이한 상태가 되며, 상기 효소와 기질 간의 친화도(affinity)도 증대될 수 있다. 상기와 같은 점을 입증하기 위하여, 야생형 M37 리파아제와 NKC가 융합된 M37 리파아제(NKC-M37 리파아제) 각각에 GFP를 융합시킨 GFP 하이브리드(GFP hybrid)를 제작하였고, 상기 리파아제의 지질 입자 내의 위치를 확인하였다(도 8).
형광 현미경 관찰 결과, NKC-M37 리파아제의 경우, 야생형 M37 리파아제에 비하여 지질 입자 내에 위치한 정도가 훨씬 컸다. 이러한 결과는, NKC와 같은 양친매성 펩타이드가 리파아제의 기능적인 형태를 형성하는 데 있어 필수적인 것임을 나타내는 결과이다.
실시예 4: 메탄올에 대한 M37 리파아제의 안정성에 미치는 양친매성 펩타이 드의 효과 확인
메탄올은 바이오디젤 생산 과정에 있어 효소 기질일 뿐만 아니라, 용매로서도 기능한다. 그러나 일반적인 리파아제(general lipase)들은 높은 농도의 메탄올 배지에서 불안정하기 때문에, 바이오디젤의 생산량은 상대적으로 제한되어 있다. 트리글리세라이드 1 당량(1 molar equivalent)을 그에 해당되는 메틸 에스터로 완전히 전화시키기 위해서는 적어도 3 당량의 메탄올이 필요하다. 그러나, 일반적인 리파아제들은 오일에 대해 1 당량의 메탄올(3.3% v/v) 이상을 포함하는 배지에서는 불활성화된다. 따라서, 바이오디젤 생산 과정은 리파아제의 불활성화를 방지하기 위하여 메탄올을 여러 단계로 추가하며 수행된다.
M37 리파아제는 높은 농도의 메탄올에서도 높은 안정도를 보이는 것으로 보고된 바 있다. 1 단계 메탄올 공급 방법(1-stepwise methanol feeding method, 3 molar equivalents)을 사용하였을 때, M37 리파아제는 70%의 전환 수율을 보인 반면, 칸디다 안탁티카(Candida antarctica) 리파아제 B인 CalB 리파아제는 매우 극 소량의 오일만을 바이오디젤로 전환시켰다.
이와 같이 높은 농도의 메탄올에서도 높은 안정도를 나타내는 M37 리파아제에 양친매성 펩타이드를 결합시키는 경우, 그 수율이 보다 증진될 수 있는지 확인하기 위하여 하기와 같이 실험을 수행하였다.
양친매성 펩타이드 중 하나인 NKC 펩타이드를 융합시킨 NKC-M37 리파아제를 사용하여 높은 농도의 메탄올에서도 안정도를 나타내는지 확인하였다. 상기 안정도 확인은 메탄올 처리후 남아있는 활성을 측정함으로써 결정하였다. CalB 리파아제, 야생형 M37 리파아제 및 NKC-M37 리파아제의 안정도를 4℃ 및 40℃에서 0%, 3.3%, 5% 및 10% 메탄올 용액에서 12시간 동안 반응시킨 후 측정하였다.
그 결과, 야생형 M37 리파아제 및 NKC-M37 리파아제는 4℃의 온도에서 3.3%, 5% 및 10% 메탄올 용액에서도 그 활성을 유지하였다. 에스테르 교환반응에서 사용되는 실제 온도인 40℃에서 메탄올에 대한 리파아제의 안정도를 측정하였을 때, NKC-M37 리파아제는 야생형 M37 리파아제에 비하여 10%의 메탄올 용액에서 안정도가 약간 감소되었으나, CalB 리파아제의 경우 빠르게 불활성화되는 결과를 나타내었다(도 9). 이러한 결과는 본 발명의 NKC-M37 리파아제는 야생형 M37 리파아제와 같이 메탄올에서 매우 안정함을 나타내는 결과로서, 본 발명의 양친매성 펩타이드-리파아제 결합체가 바이오디젤의 생산에 효율적으로 사용될 수 있음을 시사하는 결과이다.
실시예 5: 펩타이드 NKC가 융합된 M37 리파아제를 이용한 바이오디젤 생산
도 7에 나타낸 바와 같이, NKC-M37 리파아제는 야생형 M37 리파아제에 비하여 10배 높은 촉매 활성(catalytic activity)를 나타낸다. 이러한 사실은 NKC와 같은 양친매성 펩타이드가 리파아제와 지질 기질 간의 접근성을 높임으로써 M37 리파아제의 촉매 증진시키는 데 매우 효과적임을 나타내는 것이다. 이러한 점에서, 촉매 활성이 증대된 NKC-M37 리파아제를 사용하여 바이오디젤 생산 과정 및 올리브 오일의 에스테르 교환 과정을 수행하였다.
바이오디젤 생산을 위해서, 반응 플라스크에서 오일/메탄올을 화학양론적 몰비로 반응시키고, 교반하여 반응 온도를 증가시킨다. 보통, 리파아제들은 오일 대비 메탄올을 1 당량 이상을 포함하는 혼합물에서 혼합시키는 경우, 불활성화된다. 따라서, 상기에서 설명한 바와 같이 바이오디젤 과정은 메탄올을 수회 추가하여 반응시킴으로써 수행된다. 다만, 상기 실시예 4에 나타낸 결과와 같이, 야생형 M37 리파아제 및 NKC-M37 리파아제는 메탄올 3 당량 존재하에서도 안정하였다.
첫 번째로, 3 단계 메탄올 공급 방법(1 당량)을 사용하여 바이오디젤 생산을 확인하였다. 생산된 바이오디젤의 양은 가스 크로마토그래피 및 박층 크로마토크래피 방법을 사용하여 분석하였다(도 10 a).
*박층 크로마토그래피 분석 결과, 올리브 오일의 대부분은 바이오디젤로 전환되는 결과를 보였다. 도 10에는 야생형 M37 리파아제 및 NKC-M37 리파아제에 대한 95% 전환에 도달하는 시간을 나타내었다. 가스 크로마토그래피 분석 결과, 야생형 M37 리파아제를 사용하여 바이오디젤 생산을 보았을 때, 36시간에서 95%의 전환 수율을 나타낸 반면, 본 발명의 NKC-M37 리파아제는 단 15시간 만에 95%의 전환 수율을 나타내었다(도 10 a). 이러한 결과는 본 발명의 양친매성 펩타이드를 융합시킨 리파아제가 바이오디젤의 경제적이면서 효과적인 생산에 이용될 수 있음을 시사하는 결과이다.
상기에서 기술한 바와 같이, 효소적인 바이오디젤 생산 과정(Enzymatic biodisel production process)은 메탄올에 대한 리파아제의 낮은 허용도(low tolerance)를 이유로 주로 3 단계 메탄올 공급 방법이 사용된다. 다만, 경제적인 바이오디젤 생산을 위해서는 2 단계 메탄올 공급 방법이 더욱 유리하므로, 2 단계 메탄올 공급 방법을 사용하여 바이오디젤 생산을 확인하였다. 2 단계 메탄올(2 당량) 공급 방법을 사용하였을 때, 본 발명의 매우 향상된 촉매 활성을 나타내는 NKC-M37 리파아제는 21시간 만에 90% 이상의 높은 전환 수율을 나타낸 반면, 야생형 M37 리파아제는 48시간 만에 같은 수율에 도달했다(도 10 b). 이러한 결과는 본 발명의 양친매성 펩타이드가 융합된 리파아제가 야생형 리파아제에 비하여 매우 효과적인 촉매제(catalyst)임을 나타내는 결과이다. 또한, 더욱 경제적인 1 단계 메탄올 공급방법을 사용하여 바이오디젤 생산을 확인한 결과, 야생형 M37 리파아제보다 본 발명의 대표적인 결합체인 NKC-M37 리파아제가 최대 전환 수율에 훨씬 빠르게 도달하는 결과를 나타내었다(도 10 c).
또한, 바이오디젤에 드는 경비는 대개 공급 원료(feedstock) 가격에 따라 달라진다. 공급 원료 가격은 바이오디젤 생산비에 있어 70% 이상을 차지하는 것으로 알려져 있다. 높은 생산비가 드는 경우, 매우 낮은 상용성을 지니므로, 바이오디젤 가격이 석유디젤에 비하여 경쟁력을 갖추게 하기 위하여 쿠킹오일 폐기물, 우지(beef tallow), 돼지 라드(pork lard) 및 옐로우 그리이스(yellow grease)와 같은 낮은 가격의 공급 원료를 사용하기 위한 연구들이 진행 중이다. 그러나, 오일 폐기물 등을 사용하는 경우, 오일 폐기물 내에 포함되어 있는 불순물, 유리 지방산 및 함수량때문에, 바이오디젤 생산량이 급격히 감소되게 된다. M37 리파아제는 오일 폐기물 내에 있는 유리 지방산 및 함수량에 영향을 받지 않는 새로운 리파아제로 보고된 바 있어, 본 발명은 양친매성 펩타이드-리파아제 결합체도 이러한 효과를 지니는 지 확인하였다. 오일 폐기물을 기질로 하여 NKC-M37 리파아제를 사용하였을 때, 올리브 오일 및 팜유와 비슷한 정도의 바이오디젤이 생산되는 결과를 나타내었다. 이러한 결과는 본 발명의 NKC-M37 리파아제를 바이오디젤 생산에 있어 굉장한 효과를 지닌 리파아제로써 사용할 수 있음을 시사하는 결과이다.
이상의 설명으로부터, 본 발명이 속하는 기술분야의 당업자는 본 발명이 그 기술적 사상이나 필수적 특징을 변경하지 않고서 다른 구체적인 형태로 실시될 수 있다는 것을 이해할 수 있을 것이다. 이와 관련하여, 이상에서 기술한 실시 예들은 모든 면에서 예시적인 것이며 한정적인 것이 아닌 것으로서 이해해야만 한다. 본 발명의 범위는 상기 상세한 설명보다는 후술하는 특허 청구범위의 의미 및 범위 그 리고 그 등가 개념으로부터 도출되는 모든 변경 또는 변형된 형태가 본 발명의 범 위에 포함되는 것으로 해석되어야 한다.
<110> Korea Advanced Institute of Science and Technology
Intelligent Synthetic Biology Center
<120> Amphipathic peptide-lipase conjugate having advanced lipase
activity and use thereof
<130> PA120765KR-D1
<150> 10-2011-0079609
<151> 2011-08-10
<160> 91
<170> KopatentIn 2.0
<210> 1
<211> 21
<212> PRT
<213> Artificial Sequence
<220>
<223> Buforin IIb
<400> 1
Arg Ala Gly Leu Gln Phe Pro Val Gly Arg Leu Leu Arg Arg Leu Leu
1 5 10 15
Arg Arg Leu Leu Arg
20
<210> 2
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> B0
<400> 2
Arg Ala Gly Leu Gln Phe Pro Val Gly
1 5
<210> 3
<211> 19
<212> PRT
<213> Artificial Sequence
<220>
<223> Paracin I
<400> 3
Lys Gly Arg Gly Lys Gln Gly Gly Lys Val Arg Ala Lys Ala Lys Thr
1 5 10 15
Arg Ser Ser
<210> 4
<211> 19
<212> PRT
<213> Artificial Sequence
<220>
<223> NKC
<400> 4
Ala Pro Lys Ala Met Lys Leu Leu Lys Lys Leu Leu Lys Leu Gln Lys
1 5 10 15
Lys Gly Ile
<210> 5
<211> 19
<212> PRT
<213> Artificial Sequence
<220>
<223> NRC
<400> 5
Ala Pro Lys Ala Met Arg Leu Leu Arg Arg Leu Leu Arg Leu Gln Lys
1 5 10 15
Lys Gly Ile
<210> 6
<211> 476
<212> PRT
<213> Artificial Sequence
<220>
<223> lipase TliA
<400> 6
Met Gly Val Phe Asp Tyr Lys Asn Leu Gly Thr Glu Ala Ser Lys Thr
1 5 10 15
Leu Phe Ala Asp Ala Thr Ala Ile Thr Leu Tyr Thr Tyr His Asn Leu
20 25 30
Asp Asn Gly Phe Ala Val Gly Tyr Gln Gln His Gly Leu Gly Leu Gly
35 40 45
Leu Pro Ala Thr Leu Val Gly Ala Leu Leu Gly Ser Thr Asp Ser Gln
50 55 60
Gly Val Ile Pro Gly Ile Pro Trp Asn Pro Asp Ser Glu Lys Ala Ala
65 70 75 80
Leu Asp Ala Val His Ala Ala Gly Trp Thr Pro Ile Ser Ala Ser Ala
85 90 95
Leu Gly Tyr Gly Gly Lys Val Asp Ala Arg Gly Thr Phe Phe Gly Glu
100 105 110
Lys Ala Gly Tyr Thr Thr Ala Gln Ala Glu Val Leu Gly Lys Tyr Asp
115 120 125
Asp Ala Gly Lys Leu Leu Glu Ile Gly Ile Gly Phe Arg Gly Thr Ser
130 135 140
Gly Pro Arg Glu Ser Leu Ile Thr Asp Ser Ile Gly Asp Leu Val Ser
145 150 155 160
Asp Leu Leu Ala Ala Leu Gly Pro Lys Asp Tyr Ala Lys Asn Tyr Ala
165 170 175
Gly Glu Ala Phe Gly Gly Leu Leu Lys Thr Val Ala Asp Tyr Ala Gly
180 185 190
Ala His Gly Leu Ser Gly Lys Asp Val Leu Val Ser Gly His Ser Leu
195 200 205
Gly Gly Leu Ala Val Asn Ser Met Ala Asp Leu Ser Thr Ser Lys Trp
210 215 220
Ala Gly Phe Tyr Lys Asp Ala Asn Tyr Leu Ala Tyr Ala Ser Pro Thr
225 230 235 240
Gln Ser Ala Gly Asp Lys Val Leu Asn Ile Gly Tyr Glu Asn Asp Pro
245 250 255
Val Phe Arg Ala Leu Asp Gly Ser Thr Phe Asn Leu Ser Ser Leu Gly
260 265 270
Val His Asp Lys Ala His Glu Ser Thr Thr Asp Asn Ile Val Ser Phe
275 280 285
Asn Asp His Tyr Ala Ser Thr Leu Trp Asn Val Leu Pro Phe Ser Ile
290 295 300
Ala Asn Leu Ser Thr Trp Val Ser His Leu Pro Ser Ala Tyr Gly Asp
305 310 315 320
Gly Met Thr Arg Val Leu Glu Ser Gly Phe Tyr Glu Gln Met Thr Arg
325 330 335
Asp Ser Thr Ile Ile Val Ala Asn Leu Ser Asp Pro Ala Arg Ala Asn
340 345 350
Thr Trp Val Gln Asp Leu Asn Arg Asn Ala Glu Pro His Thr Gly Asn
355 360 365
Thr Phe Ile Ile Gly Ser Asp Gly Asn Asp Leu Ile Gln Gly Gly Lys
370 375 380
Gly Ala Asp Phe Ile Glu Gly Gly Lys Gly Asn Asp Thr Ile Arg Asp
385 390 395 400
Asn Ser Gly His Asn Thr Phe Leu Phe Ser Gly His Phe Gly Gln Asp
405 410 415
Arg Ile Ile Gly Tyr Gln Pro Thr Asp Arg Leu Val Phe Gln Gly Ala
420 425 430
Asp Gly Ser Thr Asp Leu Arg Asp His Ala Lys Ala Val Gly Ala Asp
435 440 445
Thr Val Leu Ser Phe Gly Ala Asp Ser Val Thr Leu Val Gly Val Gly
450 455 460
Leu Gly Gly Leu Trp Ser Glu Gly Val Leu Ile Ser
465 470 475
<210> 7
<211> 340
<212> PRT
<213> Artificial Sequence
<220>
<223> lipase M37
<400> 7
Met Ser Tyr Thr Lys Glu Gln Leu Met Leu Ala Phe Ser Tyr Met Ser
1 5 10 15
Tyr Tyr Gly Ile Thr His Thr Gly Ser Ala Lys Lys Asn Ala Glu Leu
20 25 30
Ile Leu Lys Lys Met Lys Glu Ala Leu Lys Thr Trp Lys Pro Phe Gln
35 40 45
Glu Asp Asp Trp Glu Val Val Trp Gly Pro Ala Val Tyr Thr Met Pro
50 55 60
Phe Thr Ile Phe Asn Asp Ala Met Met Tyr Val Ile Gln Lys Lys Gly
65 70 75 80
Ala Glu Gly Glu Tyr Val Ile Ala Ile Arg Gly Thr Asn Pro Val Ser
85 90 95
Ile Ser Asp Trp Leu Phe Asn Asp Phe Met Val Ser Ala Met Lys Lys
100 105 110
Trp Pro Tyr Ala Ser Val Glu Gly Arg Ile Leu Lys Ile Ser Glu Ser
115 120 125
Thr Ser Tyr Gly Leu Lys Thr Leu Gln Lys Leu Lys Pro Lys Ser His
130 135 140
Ile Pro Gly Glu Asn Lys Thr Ile Leu Gln Phe Leu Asn Glu Lys Ile
145 150 155 160
Gly Pro Glu Gly Lys Ala Lys Ile Cys Val Thr Gly His Ser Lys Gly
165 170 175
Gly Ala Leu Ser Ser Thr Leu Ala Leu Trp Leu Lys Asp Ile Gln Gly
180 185 190
Val Lys Leu Ser Gln Asn Ile Asp Ile Ser Thr Ile Pro Phe Ala Gly
195 200 205
Pro Thr Ala Gly Asn Ala Asp Phe Ala Asp Tyr Phe Asp Asp Cys Leu
210 215 220
Gly Asp Gln Cys Thr Arg Ile Ala Asn Ser Leu Asp Ile Val Pro Tyr
225 230 235 240
Ala Trp Asn Thr Asn Ser Leu Lys Lys Leu Lys Ser Ile Tyr Ile Ser
245 250 255
Glu Gln Ala Ser Val Lys Pro Leu Leu Tyr Gln Arg Ala Leu Ile Arg
260 265 270
Ala Met Ile Ala Glu Thr Lys Gly Lys Lys Tyr Lys Gln Ile Lys Ala
275 280 285
Glu Thr Pro Pro Leu Glu Gly Asn Ile Asn Pro Ile Leu Ile Glu Tyr
290 295 300
Leu Val Gln Ala Ala Tyr Gln His Val Val Gly Tyr Pro Glu Leu Met
305 310 315 320
Gly Met Met Asp Asp Ile Pro Leu Thr Asp Ile Phe Glu Asp Ala Ile
325 330 335
Ala Gly Leu Leu
340
<210> 8
<211> 497
<212> PRT
<213> Artificial Sequence
<220>
<223> Buforin IIb - lipase TliA
<400> 8
Arg Ala Gly Leu Gln Phe Pro Val Gly Arg Leu Leu Arg Arg Leu Leu
1 5 10 15
Arg Arg Leu Leu Arg Met Gly Val Phe Asp Tyr Lys Asn Leu Gly Thr
20 25 30
Glu Ala Ser Lys Thr Leu Phe Ala Asp Ala Thr Ala Ile Thr Leu Tyr
35 40 45
Thr Tyr His Asn Leu Asp Asn Gly Phe Ala Val Gly Tyr Gln Gln His
50 55 60
Gly Leu Gly Leu Gly Leu Pro Ala Thr Leu Val Gly Ala Leu Leu Gly
65 70 75 80
Ser Thr Asp Ser Gln Gly Val Ile Pro Gly Ile Pro Trp Asn Pro Asp
85 90 95
Ser Glu Lys Ala Ala Leu Asp Ala Val His Ala Ala Gly Trp Thr Pro
100 105 110
Ile Ser Ala Ser Ala Leu Gly Tyr Gly Gly Lys Val Asp Ala Arg Gly
115 120 125
Thr Phe Phe Gly Glu Lys Ala Gly Tyr Thr Thr Ala Gln Ala Glu Val
130 135 140
Leu Gly Lys Tyr Asp Asp Ala Gly Lys Leu Leu Glu Ile Gly Ile Gly
145 150 155 160
Phe Arg Gly Thr Ser Gly Pro Arg Glu Ser Leu Ile Thr Asp Ser Ile
165 170 175
Gly Asp Leu Val Ser Asp Leu Leu Ala Ala Leu Gly Pro Lys Asp Tyr
180 185 190
Ala Lys Asn Tyr Ala Gly Glu Ala Phe Gly Gly Leu Leu Lys Thr Val
195 200 205
Ala Asp Tyr Ala Gly Ala His Gly Leu Ser Gly Lys Asp Val Leu Val
210 215 220
Ser Gly His Ser Leu Gly Gly Leu Ala Val Asn Ser Met Ala Asp Leu
225 230 235 240
Ser Thr Ser Lys Trp Ala Gly Phe Tyr Lys Asp Ala Asn Tyr Leu Ala
245 250 255
Tyr Ala Ser Pro Thr Gln Ser Ala Gly Asp Lys Val Leu Asn Ile Gly
260 265 270
Tyr Glu Asn Asp Pro Val Phe Arg Ala Leu Asp Gly Ser Thr Phe Asn
275 280 285
Leu Ser Ser Leu Gly Val His Asp Lys Ala His Glu Ser Thr Thr Asp
290 295 300
Asn Ile Val Ser Phe Asn Asp His Tyr Ala Ser Thr Leu Trp Asn Val
305 310 315 320
Leu Pro Phe Ser Ile Ala Asn Leu Ser Thr Trp Val Ser His Leu Pro
325 330 335
Ser Ala Tyr Gly Asp Gly Met Thr Arg Val Leu Glu Ser Gly Phe Tyr
340 345 350
Glu Gln Met Thr Arg Asp Ser Thr Ile Ile Val Ala Asn Leu Ser Asp
355 360 365
Pro Ala Arg Ala Asn Thr Trp Val Gln Asp Leu Asn Arg Asn Ala Glu
370 375 380
Pro His Thr Gly Asn Thr Phe Ile Ile Gly Ser Asp Gly Asn Asp Leu
385 390 395 400
Ile Gln Gly Gly Lys Gly Ala Asp Phe Ile Glu Gly Gly Lys Gly Asn
405 410 415
Asp Thr Ile Arg Asp Asn Ser Gly His Asn Thr Phe Leu Phe Ser Gly
420 425 430
His Phe Gly Gln Asp Arg Ile Ile Gly Tyr Gln Pro Thr Asp Arg Leu
435 440 445
Val Phe Gln Gly Ala Asp Gly Ser Thr Asp Leu Arg Asp His Ala Lys
450 455 460
Ala Val Gly Ala Asp Thr Val Leu Ser Phe Gly Ala Asp Ser Val Thr
465 470 475 480
Leu Val Gly Val Gly Leu Gly Gly Leu Trp Ser Glu Gly Val Leu Ile
485 490 495
Ser
<210> 9
<211> 485
<212> PRT
<213> Artificial Sequence
<220>
<223> B0 - lipase TliA
<400> 9
Arg Ala Gly Leu Gln Phe Pro Val Gly Met Gly Val Phe Asp Tyr Lys
1 5 10 15
Asn Leu Gly Thr Glu Ala Ser Lys Thr Leu Phe Ala Asp Ala Thr Ala
20 25 30
Ile Thr Leu Tyr Thr Tyr His Asn Leu Asp Asn Gly Phe Ala Val Gly
35 40 45
Tyr Gln Gln His Gly Leu Gly Leu Gly Leu Pro Ala Thr Leu Val Gly
50 55 60
Ala Leu Leu Gly Ser Thr Asp Ser Gln Gly Val Ile Pro Gly Ile Pro
65 70 75 80
Trp Asn Pro Asp Ser Glu Lys Ala Ala Leu Asp Ala Val His Ala Ala
85 90 95
Gly Trp Thr Pro Ile Ser Ala Ser Ala Leu Gly Tyr Gly Gly Lys Val
100 105 110
Asp Ala Arg Gly Thr Phe Phe Gly Glu Lys Ala Gly Tyr Thr Thr Ala
115 120 125
Gln Ala Glu Val Leu Gly Lys Tyr Asp Asp Ala Gly Lys Leu Leu Glu
130 135 140
Ile Gly Ile Gly Phe Arg Gly Thr Ser Gly Pro Arg Glu Ser Leu Ile
145 150 155 160
Thr Asp Ser Ile Gly Asp Leu Val Ser Asp Leu Leu Ala Ala Leu Gly
165 170 175
Pro Lys Asp Tyr Ala Lys Asn Tyr Ala Gly Glu Ala Phe Gly Gly Leu
180 185 190
Leu Lys Thr Val Ala Asp Tyr Ala Gly Ala His Gly Leu Ser Gly Lys
195 200 205
Asp Val Leu Val Ser Gly His Ser Leu Gly Gly Leu Ala Val Asn Ser
210 215 220
Met Ala Asp Leu Ser Thr Ser Lys Trp Ala Gly Phe Tyr Lys Asp Ala
225 230 235 240
Asn Tyr Leu Ala Tyr Ala Ser Pro Thr Gln Ser Ala Gly Asp Lys Val
245 250 255
Leu Asn Ile Gly Tyr Glu Asn Asp Pro Val Phe Arg Ala Leu Asp Gly
260 265 270
Ser Thr Phe Asn Leu Ser Ser Leu Gly Val His Asp Lys Ala His Glu
275 280 285
Ser Thr Thr Asp Asn Ile Val Ser Phe Asn Asp His Tyr Ala Ser Thr
290 295 300
Leu Trp Asn Val Leu Pro Phe Ser Ile Ala Asn Leu Ser Thr Trp Val
305 310 315 320
Ser His Leu Pro Ser Ala Tyr Gly Asp Gly Met Thr Arg Val Leu Glu
325 330 335
Ser Gly Phe Tyr Glu Gln Met Thr Arg Asp Ser Thr Ile Ile Val Ala
340 345 350
Asn Leu Ser Asp Pro Ala Arg Ala Asn Thr Trp Val Gln Asp Leu Asn
355 360 365
Arg Asn Ala Glu Pro His Thr Gly Asn Thr Phe Ile Ile Gly Ser Asp
370 375 380
Gly Asn Asp Leu Ile Gln Gly Gly Lys Gly Ala Asp Phe Ile Glu Gly
385 390 395 400
Gly Lys Gly Asn Asp Thr Ile Arg Asp Asn Ser Gly His Asn Thr Phe
405 410 415
Leu Phe Ser Gly His Phe Gly Gln Asp Arg Ile Ile Gly Tyr Gln Pro
420 425 430
Thr Asp Arg Leu Val Phe Gln Gly Ala Asp Gly Ser Thr Asp Leu Arg
435 440 445
Asp His Ala Lys Ala Val Gly Ala Asp Thr Val Leu Ser Phe Gly Ala
450 455 460
Asp Ser Val Thr Leu Val Gly Val Gly Leu Gly Gly Leu Trp Ser Glu
465 470 475 480
Gly Val Leu Ile Ser
485
<210> 10
<211> 495
<212> PRT
<213> Artificial Sequence
<220>
<223> Paracin I - lipase TliA
<400> 10
Lys Gly Arg Gly Lys Gln Gly Gly Lys Val Arg Ala Lys Ala Lys Thr
1 5 10 15
Arg Ser Ser Met Gly Val Phe Asp Tyr Lys Asn Leu Gly Thr Glu Ala
20 25 30
Ser Lys Thr Leu Phe Ala Asp Ala Thr Ala Ile Thr Leu Tyr Thr Tyr
35 40 45
His Asn Leu Asp Asn Gly Phe Ala Val Gly Tyr Gln Gln His Gly Leu
50 55 60
Gly Leu Gly Leu Pro Ala Thr Leu Val Gly Ala Leu Leu Gly Ser Thr
65 70 75 80
Asp Ser Gln Gly Val Ile Pro Gly Ile Pro Trp Asn Pro Asp Ser Glu
85 90 95
Lys Ala Ala Leu Asp Ala Val His Ala Ala Gly Trp Thr Pro Ile Ser
100 105 110
Ala Ser Ala Leu Gly Tyr Gly Gly Lys Val Asp Ala Arg Gly Thr Phe
115 120 125
Phe Gly Glu Lys Ala Gly Tyr Thr Thr Ala Gln Ala Glu Val Leu Gly
130 135 140
Lys Tyr Asp Asp Ala Gly Lys Leu Leu Glu Ile Gly Ile Gly Phe Arg
145 150 155 160
Gly Thr Ser Gly Pro Arg Glu Ser Leu Ile Thr Asp Ser Ile Gly Asp
165 170 175
Leu Val Ser Asp Leu Leu Ala Ala Leu Gly Pro Lys Asp Tyr Ala Lys
180 185 190
Asn Tyr Ala Gly Glu Ala Phe Gly Gly Leu Leu Lys Thr Val Ala Asp
195 200 205
Tyr Ala Gly Ala His Gly Leu Ser Gly Lys Asp Val Leu Val Ser Gly
210 215 220
His Ser Leu Gly Gly Leu Ala Val Asn Ser Met Ala Asp Leu Ser Thr
225 230 235 240
Ser Lys Trp Ala Gly Phe Tyr Lys Asp Ala Asn Tyr Leu Ala Tyr Ala
245 250 255
Ser Pro Thr Gln Ser Ala Gly Asp Lys Val Leu Asn Ile Gly Tyr Glu
260 265 270
Asn Asp Pro Val Phe Arg Ala Leu Asp Gly Ser Thr Phe Asn Leu Ser
275 280 285
Ser Leu Gly Val His Asp Lys Ala His Glu Ser Thr Thr Asp Asn Ile
290 295 300
Val Ser Phe Asn Asp His Tyr Ala Ser Thr Leu Trp Asn Val Leu Pro
305 310 315 320
Phe Ser Ile Ala Asn Leu Ser Thr Trp Val Ser His Leu Pro Ser Ala
325 330 335
Tyr Gly Asp Gly Met Thr Arg Val Leu Glu Ser Gly Phe Tyr Glu Gln
340 345 350
Met Thr Arg Asp Ser Thr Ile Ile Val Ala Asn Leu Ser Asp Pro Ala
355 360 365
Arg Ala Asn Thr Trp Val Gln Asp Leu Asn Arg Asn Ala Glu Pro His
370 375 380
Thr Gly Asn Thr Phe Ile Ile Gly Ser Asp Gly Asn Asp Leu Ile Gln
385 390 395 400
Gly Gly Lys Gly Ala Asp Phe Ile Glu Gly Gly Lys Gly Asn Asp Thr
405 410 415
Ile Arg Asp Asn Ser Gly His Asn Thr Phe Leu Phe Ser Gly His Phe
420 425 430
Gly Gln Asp Arg Ile Ile Gly Tyr Gln Pro Thr Asp Arg Leu Val Phe
435 440 445
Gln Gly Ala Asp Gly Ser Thr Asp Leu Arg Asp His Ala Lys Ala Val
450 455 460
Gly Ala Asp Thr Val Leu Ser Phe Gly Ala Asp Ser Val Thr Leu Val
465 470 475 480
Gly Val Gly Leu Gly Gly Leu Trp Ser Glu Gly Val Leu Ile Ser
485 490 495
<210> 11
<211> 495
<212> PRT
<213> Artificial Sequence
<220>
<223> NKC - lipase TliA
<400> 11
Ala Pro Lys Ala Met Lys Leu Leu Lys Lys Leu Leu Lys Leu Gln Lys
1 5 10 15
Lys Gly Ile Met Gly Val Phe Asp Tyr Lys Asn Leu Gly Thr Glu Ala
20 25 30
Ser Lys Thr Leu Phe Ala Asp Ala Thr Ala Ile Thr Leu Tyr Thr Tyr
35 40 45
His Asn Leu Asp Asn Gly Phe Ala Val Gly Tyr Gln Gln His Gly Leu
50 55 60
Gly Leu Gly Leu Pro Ala Thr Leu Val Gly Ala Leu Leu Gly Ser Thr
65 70 75 80
Asp Ser Gln Gly Val Ile Pro Gly Ile Pro Trp Asn Pro Asp Ser Glu
85 90 95
Lys Ala Ala Leu Asp Ala Val His Ala Ala Gly Trp Thr Pro Ile Ser
100 105 110
Ala Ser Ala Leu Gly Tyr Gly Gly Lys Val Asp Ala Arg Gly Thr Phe
115 120 125
Phe Gly Glu Lys Ala Gly Tyr Thr Thr Ala Gln Ala Glu Val Leu Gly
130 135 140
Lys Tyr Asp Asp Ala Gly Lys Leu Leu Glu Ile Gly Ile Gly Phe Arg
145 150 155 160
Gly Thr Ser Gly Pro Arg Glu Ser Leu Ile Thr Asp Ser Ile Gly Asp
165 170 175
Leu Val Ser Asp Leu Leu Ala Ala Leu Gly Pro Lys Asp Tyr Ala Lys
180 185 190
Asn Tyr Ala Gly Glu Ala Phe Gly Gly Leu Leu Lys Thr Val Ala Asp
195 200 205
Tyr Ala Gly Ala His Gly Leu Ser Gly Lys Asp Val Leu Val Ser Gly
210 215 220
His Ser Leu Gly Gly Leu Ala Val Asn Ser Met Ala Asp Leu Ser Thr
225 230 235 240
Ser Lys Trp Ala Gly Phe Tyr Lys Asp Ala Asn Tyr Leu Ala Tyr Ala
245 250 255
Ser Pro Thr Gln Ser Ala Gly Asp Lys Val Leu Asn Ile Gly Tyr Glu
260 265 270
Asn Asp Pro Val Phe Arg Ala Leu Asp Gly Ser Thr Phe Asn Leu Ser
275 280 285
Ser Leu Gly Val His Asp Lys Ala His Glu Ser Thr Thr Asp Asn Ile
290 295 300
Val Ser Phe Asn Asp His Tyr Ala Ser Thr Leu Trp Asn Val Leu Pro
305 310 315 320
Phe Ser Ile Ala Asn Leu Ser Thr Trp Val Ser His Leu Pro Ser Ala
325 330 335
Tyr Gly Asp Gly Met Thr Arg Val Leu Glu Ser Gly Phe Tyr Glu Gln
340 345 350
Met Thr Arg Asp Ser Thr Ile Ile Val Ala Asn Leu Ser Asp Pro Ala
355 360 365
Arg Ala Asn Thr Trp Val Gln Asp Leu Asn Arg Asn Ala Glu Pro His
370 375 380
Thr Gly Asn Thr Phe Ile Ile Gly Ser Asp Gly Asn Asp Leu Ile Gln
385 390 395 400
Gly Gly Lys Gly Ala Asp Phe Ile Glu Gly Gly Lys Gly Asn Asp Thr
405 410 415
Ile Arg Asp Asn Ser Gly His Asn Thr Phe Leu Phe Ser Gly His Phe
420 425 430
Gly Gln Asp Arg Ile Ile Gly Tyr Gln Pro Thr Asp Arg Leu Val Phe
435 440 445
Gln Gly Ala Asp Gly Ser Thr Asp Leu Arg Asp His Ala Lys Ala Val
450 455 460
Gly Ala Asp Thr Val Leu Ser Phe Gly Ala Asp Ser Val Thr Leu Val
465 470 475 480
Gly Val Gly Leu Gly Gly Leu Trp Ser Glu Gly Val Leu Ile Ser
485 490 495
<210> 12
<211> 495
<212> PRT
<213> Artificial Sequence
<220>
<223> NRC - lipase TliA
<400> 12
Ala Pro Lys Ala Met Arg Leu Leu Arg Arg Leu Leu Arg Leu Gln Lys
1 5 10 15
Lys Gly Ile Met Gly Val Phe Asp Tyr Lys Asn Leu Gly Thr Glu Ala
20 25 30
Ser Lys Thr Leu Phe Ala Asp Ala Thr Ala Ile Thr Leu Tyr Thr Tyr
35 40 45
His Asn Leu Asp Asn Gly Phe Ala Val Gly Tyr Gln Gln His Gly Leu
50 55 60
Gly Leu Gly Leu Pro Ala Thr Leu Val Gly Ala Leu Leu Gly Ser Thr
65 70 75 80
Asp Ser Gln Gly Val Ile Pro Gly Ile Pro Trp Asn Pro Asp Ser Glu
85 90 95
Lys Ala Ala Leu Asp Ala Val His Ala Ala Gly Trp Thr Pro Ile Ser
100 105 110
Ala Ser Ala Leu Gly Tyr Gly Gly Lys Val Asp Ala Arg Gly Thr Phe
115 120 125
Phe Gly Glu Lys Ala Gly Tyr Thr Thr Ala Gln Ala Glu Val Leu Gly
130 135 140
Lys Tyr Asp Asp Ala Gly Lys Leu Leu Glu Ile Gly Ile Gly Phe Arg
145 150 155 160
Gly Thr Ser Gly Pro Arg Glu Ser Leu Ile Thr Asp Ser Ile Gly Asp
165 170 175
Leu Val Ser Asp Leu Leu Ala Ala Leu Gly Pro Lys Asp Tyr Ala Lys
180 185 190
Asn Tyr Ala Gly Glu Ala Phe Gly Gly Leu Leu Lys Thr Val Ala Asp
195 200 205
Tyr Ala Gly Ala His Gly Leu Ser Gly Lys Asp Val Leu Val Ser Gly
210 215 220
His Ser Leu Gly Gly Leu Ala Val Asn Ser Met Ala Asp Leu Ser Thr
225 230 235 240
Ser Lys Trp Ala Gly Phe Tyr Lys Asp Ala Asn Tyr Leu Ala Tyr Ala
245 250 255
Ser Pro Thr Gln Ser Ala Gly Asp Lys Val Leu Asn Ile Gly Tyr Glu
260 265 270
Asn Asp Pro Val Phe Arg Ala Leu Asp Gly Ser Thr Phe Asn Leu Ser
275 280 285
Ser Leu Gly Val His Asp Lys Ala His Glu Ser Thr Thr Asp Asn Ile
290 295 300
Val Ser Phe Asn Asp His Tyr Ala Ser Thr Leu Trp Asn Val Leu Pro
305 310 315 320
Phe Ser Ile Ala Asn Leu Ser Thr Trp Val Ser His Leu Pro Ser Ala
325 330 335
Tyr Gly Asp Gly Met Thr Arg Val Leu Glu Ser Gly Phe Tyr Glu Gln
340 345 350
Met Thr Arg Asp Ser Thr Ile Ile Val Ala Asn Leu Ser Asp Pro Ala
355 360 365
Arg Ala Asn Thr Trp Val Gln Asp Leu Asn Arg Asn Ala Glu Pro His
370 375 380
Thr Gly Asn Thr Phe Ile Ile Gly Ser Asp Gly Asn Asp Leu Ile Gln
385 390 395 400
Gly Gly Lys Gly Ala Asp Phe Ile Glu Gly Gly Lys Gly Asn Asp Thr
405 410 415
Ile Arg Asp Asn Ser Gly His Asn Thr Phe Leu Phe Ser Gly His Phe
420 425 430
Gly Gln Asp Arg Ile Ile Gly Tyr Gln Pro Thr Asp Arg Leu Val Phe
435 440 445
Gln Gly Ala Asp Gly Ser Thr Asp Leu Arg Asp His Ala Lys Ala Val
450 455 460
Gly Ala Asp Thr Val Leu Ser Phe Gly Ala Asp Ser Val Thr Leu Val
465 470 475 480
Gly Val Gly Leu Gly Gly Leu Trp Ser Glu Gly Val Leu Ile Ser
485 490 495
<210> 13
<211> 361
<212> PRT
<213> Artificial Sequence
<220>
<223> Buforin IIb - lipase M37
<400> 13
Arg Ala Gly Leu Gln Phe Pro Val Gly Arg Leu Leu Arg Arg Leu Leu
1 5 10 15
Arg Arg Leu Leu Arg Met Ser Tyr Thr Lys Glu Gln Leu Met Leu Ala
20 25 30
Phe Ser Tyr Met Ser Tyr Tyr Gly Ile Thr His Thr Gly Ser Ala Lys
35 40 45
Lys Asn Ala Glu Leu Ile Leu Lys Lys Met Lys Glu Ala Leu Lys Thr
50 55 60
Trp Lys Pro Phe Gln Glu Asp Asp Trp Glu Val Val Trp Gly Pro Ala
65 70 75 80
Val Tyr Thr Met Pro Phe Thr Ile Phe Asn Asp Ala Met Met Tyr Val
85 90 95
Ile Gln Lys Lys Gly Ala Glu Gly Glu Tyr Val Ile Ala Ile Arg Gly
100 105 110
Thr Asn Pro Val Ser Ile Ser Asp Trp Leu Phe Asn Asp Phe Met Val
115 120 125
Ser Ala Met Lys Lys Trp Pro Tyr Ala Ser Val Glu Gly Arg Ile Leu
130 135 140
Lys Ile Ser Glu Ser Thr Ser Tyr Gly Leu Lys Thr Leu Gln Lys Leu
145 150 155 160
Lys Pro Lys Ser His Ile Pro Gly Glu Asn Lys Thr Ile Leu Gln Phe
165 170 175
Leu Asn Glu Lys Ile Gly Pro Glu Gly Lys Ala Lys Ile Cys Val Thr
180 185 190
Gly His Ser Lys Gly Gly Ala Leu Ser Ser Thr Leu Ala Leu Trp Leu
195 200 205
Lys Asp Ile Gln Gly Val Lys Leu Ser Gln Asn Ile Asp Ile Ser Thr
210 215 220
Ile Pro Phe Ala Gly Pro Thr Ala Gly Asn Ala Asp Phe Ala Asp Tyr
225 230 235 240
Phe Asp Asp Cys Leu Gly Asp Gln Cys Thr Arg Ile Ala Asn Ser Leu
245 250 255
Asp Ile Val Pro Tyr Ala Trp Asn Thr Asn Ser Leu Lys Lys Leu Lys
260 265 270
Ser Ile Tyr Ile Ser Glu Gln Ala Ser Val Lys Pro Leu Leu Tyr Gln
275 280 285
Arg Ala Leu Ile Arg Ala Met Ile Ala Glu Thr Lys Gly Lys Lys Tyr
290 295 300
Lys Gln Ile Lys Ala Glu Thr Pro Pro Leu Glu Gly Asn Ile Asn Pro
305 310 315 320
Ile Leu Ile Glu Tyr Leu Val Gln Ala Ala Tyr Gln His Val Val Gly
325 330 335
Tyr Pro Glu Leu Met Gly Met Met Asp Asp Ile Pro Leu Thr Asp Ile
340 345 350
Phe Glu Asp Ala Ile Ala Gly Leu Leu
355 360
<210> 14
<211> 349
<212> PRT
<213> Artificial Sequence
<220>
<223> B0 - lipase M37
<400> 14
Arg Ala Gly Leu Gln Phe Pro Val Gly Met Ser Tyr Thr Lys Glu Gln
1 5 10 15
Leu Met Leu Ala Phe Ser Tyr Met Ser Tyr Tyr Gly Ile Thr His Thr
20 25 30
Gly Ser Ala Lys Lys Asn Ala Glu Leu Ile Leu Lys Lys Met Lys Glu
35 40 45
Ala Leu Lys Thr Trp Lys Pro Phe Gln Glu Asp Asp Trp Glu Val Val
50 55 60
Trp Gly Pro Ala Val Tyr Thr Met Pro Phe Thr Ile Phe Asn Asp Ala
65 70 75 80
Met Met Tyr Val Ile Gln Lys Lys Gly Ala Glu Gly Glu Tyr Val Ile
85 90 95
Ala Ile Arg Gly Thr Asn Pro Val Ser Ile Ser Asp Trp Leu Phe Asn
100 105 110
Asp Phe Met Val Ser Ala Met Lys Lys Trp Pro Tyr Ala Ser Val Glu
115 120 125
Gly Arg Ile Leu Lys Ile Ser Glu Ser Thr Ser Tyr Gly Leu Lys Thr
130 135 140
Leu Gln Lys Leu Lys Pro Lys Ser His Ile Pro Gly Glu Asn Lys Thr
145 150 155 160
Ile Leu Gln Phe Leu Asn Glu Lys Ile Gly Pro Glu Gly Lys Ala Lys
165 170 175
Ile Cys Val Thr Gly His Ser Lys Gly Gly Ala Leu Ser Ser Thr Leu
180 185 190
Ala Leu Trp Leu Lys Asp Ile Gln Gly Val Lys Leu Ser Gln Asn Ile
195 200 205
Asp Ile Ser Thr Ile Pro Phe Ala Gly Pro Thr Ala Gly Asn Ala Asp
210 215 220
Phe Ala Asp Tyr Phe Asp Asp Cys Leu Gly Asp Gln Cys Thr Arg Ile
225 230 235 240
Ala Asn Ser Leu Asp Ile Val Pro Tyr Ala Trp Asn Thr Asn Ser Leu
245 250 255
Lys Lys Leu Lys Ser Ile Tyr Ile Ser Glu Gln Ala Ser Val Lys Pro
260 265 270
Leu Leu Tyr Gln Arg Ala Leu Ile Arg Ala Met Ile Ala Glu Thr Lys
275 280 285
Gly Lys Lys Tyr Lys Gln Ile Lys Ala Glu Thr Pro Pro Leu Glu Gly
290 295 300
Asn Ile Asn Pro Ile Leu Ile Glu Tyr Leu Val Gln Ala Ala Tyr Gln
305 310 315 320
His Val Val Gly Tyr Pro Glu Leu Met Gly Met Met Asp Asp Ile Pro
325 330 335
Leu Thr Asp Ile Phe Glu Asp Ala Ile Ala Gly Leu Leu
340 345
<210> 15
<211> 359
<212> PRT
<213> Artificial Sequence
<220>
<223> Paracin I - lipase M37
<400> 15
Lys Gly Arg Gly Lys Gln Gly Gly Lys Val Arg Ala Lys Ala Lys Thr
1 5 10 15
Arg Ser Ser Met Ser Tyr Thr Lys Glu Gln Leu Met Leu Ala Phe Ser
20 25 30
Tyr Met Ser Tyr Tyr Gly Ile Thr His Thr Gly Ser Ala Lys Lys Asn
35 40 45
Ala Glu Leu Ile Leu Lys Lys Met Lys Glu Ala Leu Lys Thr Trp Lys
50 55 60
Pro Phe Gln Glu Asp Asp Trp Glu Val Val Trp Gly Pro Ala Val Tyr
65 70 75 80
Thr Met Pro Phe Thr Ile Phe Asn Asp Ala Met Met Tyr Val Ile Gln
85 90 95
Lys Lys Gly Ala Glu Gly Glu Tyr Val Ile Ala Ile Arg Gly Thr Asn
100 105 110
Pro Val Ser Ile Ser Asp Trp Leu Phe Asn Asp Phe Met Val Ser Ala
115 120 125
Met Lys Lys Trp Pro Tyr Ala Ser Val Glu Gly Arg Ile Leu Lys Ile
130 135 140
Ser Glu Ser Thr Ser Tyr Gly Leu Lys Thr Leu Gln Lys Leu Lys Pro
145 150 155 160
Lys Ser His Ile Pro Gly Glu Asn Lys Thr Ile Leu Gln Phe Leu Asn
165 170 175
Glu Lys Ile Gly Pro Glu Gly Lys Ala Lys Ile Cys Val Thr Gly His
180 185 190
Ser Lys Gly Gly Ala Leu Ser Ser Thr Leu Ala Leu Trp Leu Lys Asp
195 200 205
Ile Gln Gly Val Lys Leu Ser Gln Asn Ile Asp Ile Ser Thr Ile Pro
210 215 220
Phe Ala Gly Pro Thr Ala Gly Asn Ala Asp Phe Ala Asp Tyr Phe Asp
225 230 235 240
Asp Cys Leu Gly Asp Gln Cys Thr Arg Ile Ala Asn Ser Leu Asp Ile
245 250 255
Val Pro Tyr Ala Trp Asn Thr Asn Ser Leu Lys Lys Leu Lys Ser Ile
260 265 270
Tyr Ile Ser Glu Gln Ala Ser Val Lys Pro Leu Leu Tyr Gln Arg Ala
275 280 285
Leu Ile Arg Ala Met Ile Ala Glu Thr Lys Gly Lys Lys Tyr Lys Gln
290 295 300
Ile Lys Ala Glu Thr Pro Pro Leu Glu Gly Asn Ile Asn Pro Ile Leu
305 310 315 320
Ile Glu Tyr Leu Val Gln Ala Ala Tyr Gln His Val Val Gly Tyr Pro
325 330 335
Glu Leu Met Gly Met Met Asp Asp Ile Pro Leu Thr Asp Ile Phe Glu
340 345 350
Asp Ala Ile Ala Gly Leu Leu
355
<210> 16
<211> 359
<212> PRT
<213> Artificial Sequence
<220>
<223> NKC - lipase M37
<400> 16
Ala Pro Lys Ala Met Lys Leu Leu Lys Lys Leu Leu Lys Leu Gln Lys
1 5 10 15
Lys Gly Ile Met Ser Tyr Thr Lys Glu Gln Leu Met Leu Ala Phe Ser
20 25 30
Tyr Met Ser Tyr Tyr Gly Ile Thr His Thr Gly Ser Ala Lys Lys Asn
35 40 45
Ala Glu Leu Ile Leu Lys Lys Met Lys Glu Ala Leu Lys Thr Trp Lys
50 55 60
Pro Phe Gln Glu Asp Asp Trp Glu Val Val Trp Gly Pro Ala Val Tyr
65 70 75 80
Thr Met Pro Phe Thr Ile Phe Asn Asp Ala Met Met Tyr Val Ile Gln
85 90 95
Lys Lys Gly Ala Glu Gly Glu Tyr Val Ile Ala Ile Arg Gly Thr Asn
100 105 110
Pro Val Ser Ile Ser Asp Trp Leu Phe Asn Asp Phe Met Val Ser Ala
115 120 125
Met Lys Lys Trp Pro Tyr Ala Ser Val Glu Gly Arg Ile Leu Lys Ile
130 135 140
Ser Glu Ser Thr Ser Tyr Gly Leu Lys Thr Leu Gln Lys Leu Lys Pro
145 150 155 160
Lys Ser His Ile Pro Gly Glu Asn Lys Thr Ile Leu Gln Phe Leu Asn
165 170 175
Glu Lys Ile Gly Pro Glu Gly Lys Ala Lys Ile Cys Val Thr Gly His
180 185 190
Ser Lys Gly Gly Ala Leu Ser Ser Thr Leu Ala Leu Trp Leu Lys Asp
195 200 205
Ile Gln Gly Val Lys Leu Ser Gln Asn Ile Asp Ile Ser Thr Ile Pro
210 215 220
Phe Ala Gly Pro Thr Ala Gly Asn Ala Asp Phe Ala Asp Tyr Phe Asp
225 230 235 240
Asp Cys Leu Gly Asp Gln Cys Thr Arg Ile Ala Asn Ser Leu Asp Ile
245 250 255
Val Pro Tyr Ala Trp Asn Thr Asn Ser Leu Lys Lys Leu Lys Ser Ile
260 265 270
Tyr Ile Ser Glu Gln Ala Ser Val Lys Pro Leu Leu Tyr Gln Arg Ala
275 280 285
Leu Ile Arg Ala Met Ile Ala Glu Thr Lys Gly Lys Lys Tyr Lys Gln
290 295 300
Ile Lys Ala Glu Thr Pro Pro Leu Glu Gly Asn Ile Asn Pro Ile Leu
305 310 315 320
Ile Glu Tyr Leu Val Gln Ala Ala Tyr Gln His Val Val Gly Tyr Pro
325 330 335
Glu Leu Met Gly Met Met Asp Asp Ile Pro Leu Thr Asp Ile Phe Glu
340 345 350
Asp Ala Ile Ala Gly Leu Leu
355
<210> 17
<211> 359
<212> PRT
<213> Artificial Sequence
<220>
<223> NRC - lipase M37
<400> 17
Ala Pro Lys Ala Met Arg Leu Leu Arg Arg Leu Leu Arg Leu Gln Lys
1 5 10 15
Lys Gly Ile Met Ser Tyr Thr Lys Glu Gln Leu Met Leu Ala Phe Ser
20 25 30
Tyr Met Ser Tyr Tyr Gly Ile Thr His Thr Gly Ser Ala Lys Lys Asn
35 40 45
Ala Glu Leu Ile Leu Lys Lys Met Lys Glu Ala Leu Lys Thr Trp Lys
50 55 60
Pro Phe Gln Glu Asp Asp Trp Glu Val Val Trp Gly Pro Ala Val Tyr
65 70 75 80
Thr Met Pro Phe Thr Ile Phe Asn Asp Ala Met Met Tyr Val Ile Gln
85 90 95
Lys Lys Gly Ala Glu Gly Glu Tyr Val Ile Ala Ile Arg Gly Thr Asn
100 105 110
Pro Val Ser Ile Ser Asp Trp Leu Phe Asn Asp Phe Met Val Ser Ala
115 120 125
Met Lys Lys Trp Pro Tyr Ala Ser Val Glu Gly Arg Ile Leu Lys Ile
130 135 140
Ser Glu Ser Thr Ser Tyr Gly Leu Lys Thr Leu Gln Lys Leu Lys Pro
145 150 155 160
Lys Ser His Ile Pro Gly Glu Asn Lys Thr Ile Leu Gln Phe Leu Asn
165 170 175
Glu Lys Ile Gly Pro Glu Gly Lys Ala Lys Ile Cys Val Thr Gly His
180 185 190
Ser Lys Gly Gly Ala Leu Ser Ser Thr Leu Ala Leu Trp Leu Lys Asp
195 200 205
Ile Gln Gly Val Lys Leu Ser Gln Asn Ile Asp Ile Ser Thr Ile Pro
210 215 220
Phe Ala Gly Pro Thr Ala Gly Asn Ala Asp Phe Ala Asp Tyr Phe Asp
225 230 235 240
Asp Cys Leu Gly Asp Gln Cys Thr Arg Ile Ala Asn Ser Leu Asp Ile
245 250 255
Val Pro Tyr Ala Trp Asn Thr Asn Ser Leu Lys Lys Leu Lys Ser Ile
260 265 270
Tyr Ile Ser Glu Gln Ala Ser Val Lys Pro Leu Leu Tyr Gln Arg Ala
275 280 285
Leu Ile Arg Ala Met Ile Ala Glu Thr Lys Gly Lys Lys Tyr Lys Gln
290 295 300
Ile Lys Ala Glu Thr Pro Pro Leu Glu Gly Asn Ile Asn Pro Ile Leu
305 310 315 320
Ile Glu Tyr Leu Val Gln Ala Ala Tyr Gln His Val Val Gly Tyr Pro
325 330 335
Glu Leu Met Gly Met Met Asp Asp Ile Pro Leu Thr Asp Ile Phe Glu
340 345 350
Asp Ala Ile Ala Gly Leu Leu
355
<210> 18
<211> 497
<212> PRT
<213> Artificial Sequence
<220>
<223> lipase TliA - Buforin IIb
<400> 18
Met Gly Val Phe Asp Tyr Lys Asn Leu Gly Thr Glu Ala Ser Lys Thr
1 5 10 15
Leu Phe Ala Asp Ala Thr Ala Ile Thr Leu Tyr Thr Tyr His Asn Leu
20 25 30
Asp Asn Gly Phe Ala Val Gly Tyr Gln Gln His Gly Leu Gly Leu Gly
35 40 45
Leu Pro Ala Thr Leu Val Gly Ala Leu Leu Gly Ser Thr Asp Ser Gln
50 55 60
Gly Val Ile Pro Gly Ile Pro Trp Asn Pro Asp Ser Glu Lys Ala Ala
65 70 75 80
Leu Asp Ala Val His Ala Ala Gly Trp Thr Pro Ile Ser Ala Ser Ala
85 90 95
Leu Gly Tyr Gly Gly Lys Val Asp Ala Arg Gly Thr Phe Phe Gly Glu
100 105 110
Lys Ala Gly Tyr Thr Thr Ala Gln Ala Glu Val Leu Gly Lys Tyr Asp
115 120 125
Asp Ala Gly Lys Leu Leu Glu Ile Gly Ile Gly Phe Arg Gly Thr Ser
130 135 140
Gly Pro Arg Glu Ser Leu Ile Thr Asp Ser Ile Gly Asp Leu Val Ser
145 150 155 160
Asp Leu Leu Ala Ala Leu Gly Pro Lys Asp Tyr Ala Lys Asn Tyr Ala
165 170 175
Gly Glu Ala Phe Gly Gly Leu Leu Lys Thr Val Ala Asp Tyr Ala Gly
180 185 190
Ala His Gly Leu Ser Gly Lys Asp Val Leu Val Ser Gly His Ser Leu
195 200 205
Gly Gly Leu Ala Val Asn Ser Met Ala Asp Leu Ser Thr Ser Lys Trp
210 215 220
Ala Gly Phe Tyr Lys Asp Ala Asn Tyr Leu Ala Tyr Ala Ser Pro Thr
225 230 235 240
Gln Ser Ala Gly Asp Lys Val Leu Asn Ile Gly Tyr Glu Asn Asp Pro
245 250 255
Val Phe Arg Ala Leu Asp Gly Ser Thr Phe Asn Leu Ser Ser Leu Gly
260 265 270
Val His Asp Lys Ala His Glu Ser Thr Thr Asp Asn Ile Val Ser Phe
275 280 285
Asn Asp His Tyr Ala Ser Thr Leu Trp Asn Val Leu Pro Phe Ser Ile
290 295 300
Ala Asn Leu Ser Thr Trp Val Ser His Leu Pro Ser Ala Tyr Gly Asp
305 310 315 320
Gly Met Thr Arg Val Leu Glu Ser Gly Phe Tyr Glu Gln Met Thr Arg
325 330 335
Asp Ser Thr Ile Ile Val Ala Asn Leu Ser Asp Pro Ala Arg Ala Asn
340 345 350
Thr Trp Val Gln Asp Leu Asn Arg Asn Ala Glu Pro His Thr Gly Asn
355 360 365
Thr Phe Ile Ile Gly Ser Asp Gly Asn Asp Leu Ile Gln Gly Gly Lys
370 375 380
Gly Ala Asp Phe Ile Glu Gly Gly Lys Gly Asn Asp Thr Ile Arg Asp
385 390 395 400
Asn Ser Gly His Asn Thr Phe Leu Phe Ser Gly His Phe Gly Gln Asp
405 410 415
Arg Ile Ile Gly Tyr Gln Pro Thr Asp Arg Leu Val Phe Gln Gly Ala
420 425 430
Asp Gly Ser Thr Asp Leu Arg Asp His Ala Lys Ala Val Gly Ala Asp
435 440 445
Thr Val Leu Ser Phe Gly Ala Asp Ser Val Thr Leu Val Gly Val Gly
450 455 460
Leu Gly Gly Leu Trp Ser Glu Gly Val Leu Ile Ser Arg Ala Gly Leu
465 470 475 480
Gln Phe Pro Val Gly Arg Leu Leu Arg Arg Leu Leu Arg Arg Leu Leu
485 490 495
Arg
<210> 19
<211> 485
<212> PRT
<213> Artificial Sequence
<220>
<223> lipase TliA - B0
<400> 19
Met Gly Val Phe Asp Tyr Lys Asn Leu Gly Thr Glu Ala Ser Lys Thr
1 5 10 15
Leu Phe Ala Asp Ala Thr Ala Ile Thr Leu Tyr Thr Tyr His Asn Leu
20 25 30
Asp Asn Gly Phe Ala Val Gly Tyr Gln Gln His Gly Leu Gly Leu Gly
35 40 45
Leu Pro Ala Thr Leu Val Gly Ala Leu Leu Gly Ser Thr Asp Ser Gln
50 55 60
Gly Val Ile Pro Gly Ile Pro Trp Asn Pro Asp Ser Glu Lys Ala Ala
65 70 75 80
Leu Asp Ala Val His Ala Ala Gly Trp Thr Pro Ile Ser Ala Ser Ala
85 90 95
Leu Gly Tyr Gly Gly Lys Val Asp Ala Arg Gly Thr Phe Phe Gly Glu
100 105 110
Lys Ala Gly Tyr Thr Thr Ala Gln Ala Glu Val Leu Gly Lys Tyr Asp
115 120 125
Asp Ala Gly Lys Leu Leu Glu Ile Gly Ile Gly Phe Arg Gly Thr Ser
130 135 140
Gly Pro Arg Glu Ser Leu Ile Thr Asp Ser Ile Gly Asp Leu Val Ser
145 150 155 160
Asp Leu Leu Ala Ala Leu Gly Pro Lys Asp Tyr Ala Lys Asn Tyr Ala
165 170 175
Gly Glu Ala Phe Gly Gly Leu Leu Lys Thr Val Ala Asp Tyr Ala Gly
180 185 190
Ala His Gly Leu Ser Gly Lys Asp Val Leu Val Ser Gly His Ser Leu
195 200 205
Gly Gly Leu Ala Val Asn Ser Met Ala Asp Leu Ser Thr Ser Lys Trp
210 215 220
Ala Gly Phe Tyr Lys Asp Ala Asn Tyr Leu Ala Tyr Ala Ser Pro Thr
225 230 235 240
Gln Ser Ala Gly Asp Lys Val Leu Asn Ile Gly Tyr Glu Asn Asp Pro
245 250 255
Val Phe Arg Ala Leu Asp Gly Ser Thr Phe Asn Leu Ser Ser Leu Gly
260 265 270
Val His Asp Lys Ala His Glu Ser Thr Thr Asp Asn Ile Val Ser Phe
275 280 285
Asn Asp His Tyr Ala Ser Thr Leu Trp Asn Val Leu Pro Phe Ser Ile
290 295 300
Ala Asn Leu Ser Thr Trp Val Ser His Leu Pro Ser Ala Tyr Gly Asp
305 310 315 320
Gly Met Thr Arg Val Leu Glu Ser Gly Phe Tyr Glu Gln Met Thr Arg
325 330 335
Asp Ser Thr Ile Ile Val Ala Asn Leu Ser Asp Pro Ala Arg Ala Asn
340 345 350
Thr Trp Val Gln Asp Leu Asn Arg Asn Ala Glu Pro His Thr Gly Asn
355 360 365
Thr Phe Ile Ile Gly Ser Asp Gly Asn Asp Leu Ile Gln Gly Gly Lys
370 375 380
Gly Ala Asp Phe Ile Glu Gly Gly Lys Gly Asn Asp Thr Ile Arg Asp
385 390 395 400
Asn Ser Gly His Asn Thr Phe Leu Phe Ser Gly His Phe Gly Gln Asp
405 410 415
Arg Ile Ile Gly Tyr Gln Pro Thr Asp Arg Leu Val Phe Gln Gly Ala
420 425 430
Asp Gly Ser Thr Asp Leu Arg Asp His Ala Lys Ala Val Gly Ala Asp
435 440 445
Thr Val Leu Ser Phe Gly Ala Asp Ser Val Thr Leu Val Gly Val Gly
450 455 460
Leu Gly Gly Leu Trp Ser Glu Gly Val Leu Ile Ser Arg Ala Gly Leu
465 470 475 480
Gln Phe Pro Val Gly
485
<210> 20
<211> 495
<212> PRT
<213> Artificial Sequence
<220>
<223> lipase TliA - Paracin I
<400> 20
Met Gly Val Phe Asp Tyr Lys Asn Leu Gly Thr Glu Ala Ser Lys Thr
1 5 10 15
Leu Phe Ala Asp Ala Thr Ala Ile Thr Leu Tyr Thr Tyr His Asn Leu
20 25 30
Asp Asn Gly Phe Ala Val Gly Tyr Gln Gln His Gly Leu Gly Leu Gly
35 40 45
Leu Pro Ala Thr Leu Val Gly Ala Leu Leu Gly Ser Thr Asp Ser Gln
50 55 60
Gly Val Ile Pro Gly Ile Pro Trp Asn Pro Asp Ser Glu Lys Ala Ala
65 70 75 80
Leu Asp Ala Val His Ala Ala Gly Trp Thr Pro Ile Ser Ala Ser Ala
85 90 95
Leu Gly Tyr Gly Gly Lys Val Asp Ala Arg Gly Thr Phe Phe Gly Glu
100 105 110
Lys Ala Gly Tyr Thr Thr Ala Gln Ala Glu Val Leu Gly Lys Tyr Asp
115 120 125
Asp Ala Gly Lys Leu Leu Glu Ile Gly Ile Gly Phe Arg Gly Thr Ser
130 135 140
Gly Pro Arg Glu Ser Leu Ile Thr Asp Ser Ile Gly Asp Leu Val Ser
145 150 155 160
Asp Leu Leu Ala Ala Leu Gly Pro Lys Asp Tyr Ala Lys Asn Tyr Ala
165 170 175
Gly Glu Ala Phe Gly Gly Leu Leu Lys Thr Val Ala Asp Tyr Ala Gly
180 185 190
Ala His Gly Leu Ser Gly Lys Asp Val Leu Val Ser Gly His Ser Leu
195 200 205
Gly Gly Leu Ala Val Asn Ser Met Ala Asp Leu Ser Thr Ser Lys Trp
210 215 220
Ala Gly Phe Tyr Lys Asp Ala Asn Tyr Leu Ala Tyr Ala Ser Pro Thr
225 230 235 240
Gln Ser Ala Gly Asp Lys Val Leu Asn Ile Gly Tyr Glu Asn Asp Pro
245 250 255
Val Phe Arg Ala Leu Asp Gly Ser Thr Phe Asn Leu Ser Ser Leu Gly
260 265 270
Val His Asp Lys Ala His Glu Ser Thr Thr Asp Asn Ile Val Ser Phe
275 280 285
Asn Asp His Tyr Ala Ser Thr Leu Trp Asn Val Leu Pro Phe Ser Ile
290 295 300
Ala Asn Leu Ser Thr Trp Val Ser His Leu Pro Ser Ala Tyr Gly Asp
305 310 315 320
Gly Met Thr Arg Val Leu Glu Ser Gly Phe Tyr Glu Gln Met Thr Arg
325 330 335
Asp Ser Thr Ile Ile Val Ala Asn Leu Ser Asp Pro Ala Arg Ala Asn
340 345 350
Thr Trp Val Gln Asp Leu Asn Arg Asn Ala Glu Pro His Thr Gly Asn
355 360 365
Thr Phe Ile Ile Gly Ser Asp Gly Asn Asp Leu Ile Gln Gly Gly Lys
370 375 380
Gly Ala Asp Phe Ile Glu Gly Gly Lys Gly Asn Asp Thr Ile Arg Asp
385 390 395 400
Asn Ser Gly His Asn Thr Phe Leu Phe Ser Gly His Phe Gly Gln Asp
405 410 415
Arg Ile Ile Gly Tyr Gln Pro Thr Asp Arg Leu Val Phe Gln Gly Ala
420 425 430
Asp Gly Ser Thr Asp Leu Arg Asp His Ala Lys Ala Val Gly Ala Asp
435 440 445
Thr Val Leu Ser Phe Gly Ala Asp Ser Val Thr Leu Val Gly Val Gly
450 455 460
Leu Gly Gly Leu Trp Ser Glu Gly Val Leu Ile Ser Lys Gly Arg Gly
465 470 475 480
Lys Gln Gly Gly Lys Val Arg Ala Lys Ala Lys Thr Arg Ser Ser
485 490 495
<210> 21
<211> 495
<212> PRT
<213> Artificial Sequence
<220>
<223> lipase TliA - NKC
<400> 21
Met Gly Val Phe Asp Tyr Lys Asn Leu Gly Thr Glu Ala Ser Lys Thr
1 5 10 15
Leu Phe Ala Asp Ala Thr Ala Ile Thr Leu Tyr Thr Tyr His Asn Leu
20 25 30
Asp Asn Gly Phe Ala Val Gly Tyr Gln Gln His Gly Leu Gly Leu Gly
35 40 45
Leu Pro Ala Thr Leu Val Gly Ala Leu Leu Gly Ser Thr Asp Ser Gln
50 55 60
Gly Val Ile Pro Gly Ile Pro Trp Asn Pro Asp Ser Glu Lys Ala Ala
65 70 75 80
Leu Asp Ala Val His Ala Ala Gly Trp Thr Pro Ile Ser Ala Ser Ala
85 90 95
Leu Gly Tyr Gly Gly Lys Val Asp Ala Arg Gly Thr Phe Phe Gly Glu
100 105 110
Lys Ala Gly Tyr Thr Thr Ala Gln Ala Glu Val Leu Gly Lys Tyr Asp
115 120 125
Asp Ala Gly Lys Leu Leu Glu Ile Gly Ile Gly Phe Arg Gly Thr Ser
130 135 140
Gly Pro Arg Glu Ser Leu Ile Thr Asp Ser Ile Gly Asp Leu Val Ser
145 150 155 160
Asp Leu Leu Ala Ala Leu Gly Pro Lys Asp Tyr Ala Lys Asn Tyr Ala
165 170 175
Gly Glu Ala Phe Gly Gly Leu Leu Lys Thr Val Ala Asp Tyr Ala Gly
180 185 190
Ala His Gly Leu Ser Gly Lys Asp Val Leu Val Ser Gly His Ser Leu
195 200 205
Gly Gly Leu Ala Val Asn Ser Met Ala Asp Leu Ser Thr Ser Lys Trp
210 215 220
Ala Gly Phe Tyr Lys Asp Ala Asn Tyr Leu Ala Tyr Ala Ser Pro Thr
225 230 235 240
Gln Ser Ala Gly Asp Lys Val Leu Asn Ile Gly Tyr Glu Asn Asp Pro
245 250 255
Val Phe Arg Ala Leu Asp Gly Ser Thr Phe Asn Leu Ser Ser Leu Gly
260 265 270
Val His Asp Lys Ala His Glu Ser Thr Thr Asp Asn Ile Val Ser Phe
275 280 285
Asn Asp His Tyr Ala Ser Thr Leu Trp Asn Val Leu Pro Phe Ser Ile
290 295 300
Ala Asn Leu Ser Thr Trp Val Ser His Leu Pro Ser Ala Tyr Gly Asp
305 310 315 320
Gly Met Thr Arg Val Leu Glu Ser Gly Phe Tyr Glu Gln Met Thr Arg
325 330 335
Asp Ser Thr Ile Ile Val Ala Asn Leu Ser Asp Pro Ala Arg Ala Asn
340 345 350
Thr Trp Val Gln Asp Leu Asn Arg Asn Ala Glu Pro His Thr Gly Asn
355 360 365
Thr Phe Ile Ile Gly Ser Asp Gly Asn Asp Leu Ile Gln Gly Gly Lys
370 375 380
Gly Ala Asp Phe Ile Glu Gly Gly Lys Gly Asn Asp Thr Ile Arg Asp
385 390 395 400
Asn Ser Gly His Asn Thr Phe Leu Phe Ser Gly His Phe Gly Gln Asp
405 410 415
Arg Ile Ile Gly Tyr Gln Pro Thr Asp Arg Leu Val Phe Gln Gly Ala
420 425 430
Asp Gly Ser Thr Asp Leu Arg Asp His Ala Lys Ala Val Gly Ala Asp
435 440 445
Thr Val Leu Ser Phe Gly Ala Asp Ser Val Thr Leu Val Gly Val Gly
450 455 460
Leu Gly Gly Leu Trp Ser Glu Gly Val Leu Ile Ser Ala Pro Lys Ala
465 470 475 480
Met Lys Leu Leu Lys Lys Leu Leu Lys Leu Gln Lys Lys Gly Ile
485 490 495
<210> 22
<211> 495
<212> PRT
<213> Artificial Sequence
<220>
<223> lipase TliA - NRC
<400> 22
Met Gly Val Phe Asp Tyr Lys Asn Leu Gly Thr Glu Ala Ser Lys Thr
1 5 10 15
Leu Phe Ala Asp Ala Thr Ala Ile Thr Leu Tyr Thr Tyr His Asn Leu
20 25 30
Asp Asn Gly Phe Ala Val Gly Tyr Gln Gln His Gly Leu Gly Leu Gly
35 40 45
Leu Pro Ala Thr Leu Val Gly Ala Leu Leu Gly Ser Thr Asp Ser Gln
50 55 60
Gly Val Ile Pro Gly Ile Pro Trp Asn Pro Asp Ser Glu Lys Ala Ala
65 70 75 80
Leu Asp Ala Val His Ala Ala Gly Trp Thr Pro Ile Ser Ala Ser Ala
85 90 95
Leu Gly Tyr Gly Gly Lys Val Asp Ala Arg Gly Thr Phe Phe Gly Glu
100 105 110
Lys Ala Gly Tyr Thr Thr Ala Gln Ala Glu Val Leu Gly Lys Tyr Asp
115 120 125
Asp Ala Gly Lys Leu Leu Glu Ile Gly Ile Gly Phe Arg Gly Thr Ser
130 135 140
Gly Pro Arg Glu Ser Leu Ile Thr Asp Ser Ile Gly Asp Leu Val Ser
145 150 155 160
Asp Leu Leu Ala Ala Leu Gly Pro Lys Asp Tyr Ala Lys Asn Tyr Ala
165 170 175
Gly Glu Ala Phe Gly Gly Leu Leu Lys Thr Val Ala Asp Tyr Ala Gly
180 185 190
Ala His Gly Leu Ser Gly Lys Asp Val Leu Val Ser Gly His Ser Leu
195 200 205
Gly Gly Leu Ala Val Asn Ser Met Ala Asp Leu Ser Thr Ser Lys Trp
210 215 220
Ala Gly Phe Tyr Lys Asp Ala Asn Tyr Leu Ala Tyr Ala Ser Pro Thr
225 230 235 240
Gln Ser Ala Gly Asp Lys Val Leu Asn Ile Gly Tyr Glu Asn Asp Pro
245 250 255
Val Phe Arg Ala Leu Asp Gly Ser Thr Phe Asn Leu Ser Ser Leu Gly
260 265 270
Val His Asp Lys Ala His Glu Ser Thr Thr Asp Asn Ile Val Ser Phe
275 280 285
Asn Asp His Tyr Ala Ser Thr Leu Trp Asn Val Leu Pro Phe Ser Ile
290 295 300
Ala Asn Leu Ser Thr Trp Val Ser His Leu Pro Ser Ala Tyr Gly Asp
305 310 315 320
Gly Met Thr Arg Val Leu Glu Ser Gly Phe Tyr Glu Gln Met Thr Arg
325 330 335
Asp Ser Thr Ile Ile Val Ala Asn Leu Ser Asp Pro Ala Arg Ala Asn
340 345 350
Thr Trp Val Gln Asp Leu Asn Arg Asn Ala Glu Pro His Thr Gly Asn
355 360 365
Thr Phe Ile Ile Gly Ser Asp Gly Asn Asp Leu Ile Gln Gly Gly Lys
370 375 380
Gly Ala Asp Phe Ile Glu Gly Gly Lys Gly Asn Asp Thr Ile Arg Asp
385 390 395 400
Asn Ser Gly His Asn Thr Phe Leu Phe Ser Gly His Phe Gly Gln Asp
405 410 415
Arg Ile Ile Gly Tyr Gln Pro Thr Asp Arg Leu Val Phe Gln Gly Ala
420 425 430
Asp Gly Ser Thr Asp Leu Arg Asp His Ala Lys Ala Val Gly Ala Asp
435 440 445
Thr Val Leu Ser Phe Gly Ala Asp Ser Val Thr Leu Val Gly Val Gly
450 455 460
Leu Gly Gly Leu Trp Ser Glu Gly Val Leu Ile Ser Ala Pro Lys Ala
465 470 475 480
Met Arg Leu Leu Arg Arg Leu Leu Arg Leu Gln Lys Lys Gly Ile
485 490 495
<210> 23
<211> 361
<212> PRT
<213> Artificial Sequence
<220>
<223> lipase M37- Buforin IIb
<400> 23
Met Ser Tyr Thr Lys Glu Gln Leu Met Leu Ala Phe Ser Tyr Met Ser
1 5 10 15
Tyr Tyr Gly Ile Thr His Thr Gly Ser Ala Lys Lys Asn Ala Glu Leu
20 25 30
Ile Leu Lys Lys Met Lys Glu Ala Leu Lys Thr Trp Lys Pro Phe Gln
35 40 45
Glu Asp Asp Trp Glu Val Val Trp Gly Pro Ala Val Tyr Thr Met Pro
50 55 60
Phe Thr Ile Phe Asn Asp Ala Met Met Tyr Val Ile Gln Lys Lys Gly
65 70 75 80
Ala Glu Gly Glu Tyr Val Ile Ala Ile Arg Gly Thr Asn Pro Val Ser
85 90 95
Ile Ser Asp Trp Leu Phe Asn Asp Phe Met Val Ser Ala Met Lys Lys
100 105 110
Trp Pro Tyr Ala Ser Val Glu Gly Arg Ile Leu Lys Ile Ser Glu Ser
115 120 125
Thr Ser Tyr Gly Leu Lys Thr Leu Gln Lys Leu Lys Pro Lys Ser His
130 135 140
Ile Pro Gly Glu Asn Lys Thr Ile Leu Gln Phe Leu Asn Glu Lys Ile
145 150 155 160
Gly Pro Glu Gly Lys Ala Lys Ile Cys Val Thr Gly His Ser Lys Gly
165 170 175
Gly Ala Leu Ser Ser Thr Leu Ala Leu Trp Leu Lys Asp Ile Gln Gly
180 185 190
Val Lys Leu Ser Gln Asn Ile Asp Ile Ser Thr Ile Pro Phe Ala Gly
195 200 205
Pro Thr Ala Gly Asn Ala Asp Phe Ala Asp Tyr Phe Asp Asp Cys Leu
210 215 220
Gly Asp Gln Cys Thr Arg Ile Ala Asn Ser Leu Asp Ile Val Pro Tyr
225 230 235 240
Ala Trp Asn Thr Asn Ser Leu Lys Lys Leu Lys Ser Ile Tyr Ile Ser
245 250 255
Glu Gln Ala Ser Val Lys Pro Leu Leu Tyr Gln Arg Ala Leu Ile Arg
260 265 270
Ala Met Ile Ala Glu Thr Lys Gly Lys Lys Tyr Lys Gln Ile Lys Ala
275 280 285
Glu Thr Pro Pro Leu Glu Gly Asn Ile Asn Pro Ile Leu Ile Glu Tyr
290 295 300
Leu Val Gln Ala Ala Tyr Gln His Val Val Gly Tyr Pro Glu Leu Met
305 310 315 320
Gly Met Met Asp Asp Ile Pro Leu Thr Asp Ile Phe Glu Asp Ala Ile
325 330 335
Ala Gly Leu Leu Arg Ala Gly Leu Gln Phe Pro Val Gly Arg Leu Leu
340 345 350
Arg Arg Leu Leu Arg Arg Leu Leu Arg
355 360
<210> 24
<211> 349
<212> PRT
<213> Artificial Sequence
<220>
<223> lipase M37 - B0
<400> 24
Met Ser Tyr Thr Lys Glu Gln Leu Met Leu Ala Phe Ser Tyr Met Ser
1 5 10 15
Tyr Tyr Gly Ile Thr His Thr Gly Ser Ala Lys Lys Asn Ala Glu Leu
20 25 30
Ile Leu Lys Lys Met Lys Glu Ala Leu Lys Thr Trp Lys Pro Phe Gln
35 40 45
Glu Asp Asp Trp Glu Val Val Trp Gly Pro Ala Val Tyr Thr Met Pro
50 55 60
Phe Thr Ile Phe Asn Asp Ala Met Met Tyr Val Ile Gln Lys Lys Gly
65 70 75 80
Ala Glu Gly Glu Tyr Val Ile Ala Ile Arg Gly Thr Asn Pro Val Ser
85 90 95
Ile Ser Asp Trp Leu Phe Asn Asp Phe Met Val Ser Ala Met Lys Lys
100 105 110
Trp Pro Tyr Ala Ser Val Glu Gly Arg Ile Leu Lys Ile Ser Glu Ser
115 120 125
Thr Ser Tyr Gly Leu Lys Thr Leu Gln Lys Leu Lys Pro Lys Ser His
130 135 140
Ile Pro Gly Glu Asn Lys Thr Ile Leu Gln Phe Leu Asn Glu Lys Ile
145 150 155 160
Gly Pro Glu Gly Lys Ala Lys Ile Cys Val Thr Gly His Ser Lys Gly
165 170 175
Gly Ala Leu Ser Ser Thr Leu Ala Leu Trp Leu Lys Asp Ile Gln Gly
180 185 190
Val Lys Leu Ser Gln Asn Ile Asp Ile Ser Thr Ile Pro Phe Ala Gly
195 200 205
Pro Thr Ala Gly Asn Ala Asp Phe Ala Asp Tyr Phe Asp Asp Cys Leu
210 215 220
Gly Asp Gln Cys Thr Arg Ile Ala Asn Ser Leu Asp Ile Val Pro Tyr
225 230 235 240
Ala Trp Asn Thr Asn Ser Leu Lys Lys Leu Lys Ser Ile Tyr Ile Ser
245 250 255
Glu Gln Ala Ser Val Lys Pro Leu Leu Tyr Gln Arg Ala Leu Ile Arg
260 265 270
Ala Met Ile Ala Glu Thr Lys Gly Lys Lys Tyr Lys Gln Ile Lys Ala
275 280 285
Glu Thr Pro Pro Leu Glu Gly Asn Ile Asn Pro Ile Leu Ile Glu Tyr
290 295 300
Leu Val Gln Ala Ala Tyr Gln His Val Val Gly Tyr Pro Glu Leu Met
305 310 315 320
Gly Met Met Asp Asp Ile Pro Leu Thr Asp Ile Phe Glu Asp Ala Ile
325 330 335
Ala Gly Leu Leu Arg Ala Gly Leu Gln Phe Pro Val Gly
340 345
<210> 25
<211> 359
<212> PRT
<213> Artificial Sequence
<220>
<223> lipase M37 - Paracin I
<400> 25
Met Ser Tyr Thr Lys Glu Gln Leu Met Leu Ala Phe Ser Tyr Met Ser
1 5 10 15
Tyr Tyr Gly Ile Thr His Thr Gly Ser Ala Lys Lys Asn Ala Glu Leu
20 25 30
Ile Leu Lys Lys Met Lys Glu Ala Leu Lys Thr Trp Lys Pro Phe Gln
35 40 45
Glu Asp Asp Trp Glu Val Val Trp Gly Pro Ala Val Tyr Thr Met Pro
50 55 60
Phe Thr Ile Phe Asn Asp Ala Met Met Tyr Val Ile Gln Lys Lys Gly
65 70 75 80
Ala Glu Gly Glu Tyr Val Ile Ala Ile Arg Gly Thr Asn Pro Val Ser
85 90 95
Ile Ser Asp Trp Leu Phe Asn Asp Phe Met Val Ser Ala Met Lys Lys
100 105 110
Trp Pro Tyr Ala Ser Val Glu Gly Arg Ile Leu Lys Ile Ser Glu Ser
115 120 125
Thr Ser Tyr Gly Leu Lys Thr Leu Gln Lys Leu Lys Pro Lys Ser His
130 135 140
Ile Pro Gly Glu Asn Lys Thr Ile Leu Gln Phe Leu Asn Glu Lys Ile
145 150 155 160
Gly Pro Glu Gly Lys Ala Lys Ile Cys Val Thr Gly His Ser Lys Gly
165 170 175
Gly Ala Leu Ser Ser Thr Leu Ala Leu Trp Leu Lys Asp Ile Gln Gly
180 185 190
Val Lys Leu Ser Gln Asn Ile Asp Ile Ser Thr Ile Pro Phe Ala Gly
195 200 205
Pro Thr Ala Gly Asn Ala Asp Phe Ala Asp Tyr Phe Asp Asp Cys Leu
210 215 220
Gly Asp Gln Cys Thr Arg Ile Ala Asn Ser Leu Asp Ile Val Pro Tyr
225 230 235 240
Ala Trp Asn Thr Asn Ser Leu Lys Lys Leu Lys Ser Ile Tyr Ile Ser
245 250 255
Glu Gln Ala Ser Val Lys Pro Leu Leu Tyr Gln Arg Ala Leu Ile Arg
260 265 270
Ala Met Ile Ala Glu Thr Lys Gly Lys Lys Tyr Lys Gln Ile Lys Ala
275 280 285
Glu Thr Pro Pro Leu Glu Gly Asn Ile Asn Pro Ile Leu Ile Glu Tyr
290 295 300
Leu Val Gln Ala Ala Tyr Gln His Val Val Gly Tyr Pro Glu Leu Met
305 310 315 320
Gly Met Met Asp Asp Ile Pro Leu Thr Asp Ile Phe Glu Asp Ala Ile
325 330 335
Ala Gly Leu Leu Lys Gly Arg Gly Lys Gln Gly Gly Lys Val Arg Ala
340 345 350
Lys Ala Lys Thr Arg Ser Ser
355
<210> 26
<211> 359
<212> PRT
<213> Artificial Sequence
<220>
<223> lipase M37 - NKC
<400> 26
Met Ser Tyr Thr Lys Glu Gln Leu Met Leu Ala Phe Ser Tyr Met Ser
1 5 10 15
Tyr Tyr Gly Ile Thr His Thr Gly Ser Ala Lys Lys Asn Ala Glu Leu
20 25 30
Ile Leu Lys Lys Met Lys Glu Ala Leu Lys Thr Trp Lys Pro Phe Gln
35 40 45
Glu Asp Asp Trp Glu Val Val Trp Gly Pro Ala Val Tyr Thr Met Pro
50 55 60
Phe Thr Ile Phe Asn Asp Ala Met Met Tyr Val Ile Gln Lys Lys Gly
65 70 75 80
Ala Glu Gly Glu Tyr Val Ile Ala Ile Arg Gly Thr Asn Pro Val Ser
85 90 95
Ile Ser Asp Trp Leu Phe Asn Asp Phe Met Val Ser Ala Met Lys Lys
100 105 110
Trp Pro Tyr Ala Ser Val Glu Gly Arg Ile Leu Lys Ile Ser Glu Ser
115 120 125
Thr Ser Tyr Gly Leu Lys Thr Leu Gln Lys Leu Lys Pro Lys Ser His
130 135 140
Ile Pro Gly Glu Asn Lys Thr Ile Leu Gln Phe Leu Asn Glu Lys Ile
145 150 155 160
Gly Pro Glu Gly Lys Ala Lys Ile Cys Val Thr Gly His Ser Lys Gly
165 170 175
Gly Ala Leu Ser Ser Thr Leu Ala Leu Trp Leu Lys Asp Ile Gln Gly
180 185 190
Val Lys Leu Ser Gln Asn Ile Asp Ile Ser Thr Ile Pro Phe Ala Gly
195 200 205
Pro Thr Ala Gly Asn Ala Asp Phe Ala Asp Tyr Phe Asp Asp Cys Leu
210 215 220
Gly Asp Gln Cys Thr Arg Ile Ala Asn Ser Leu Asp Ile Val Pro Tyr
225 230 235 240
Ala Trp Asn Thr Asn Ser Leu Lys Lys Leu Lys Ser Ile Tyr Ile Ser
245 250 255
Glu Gln Ala Ser Val Lys Pro Leu Leu Tyr Gln Arg Ala Leu Ile Arg
260 265 270
Ala Met Ile Ala Glu Thr Lys Gly Lys Lys Tyr Lys Gln Ile Lys Ala
275 280 285
Glu Thr Pro Pro Leu Glu Gly Asn Ile Asn Pro Ile Leu Ile Glu Tyr
290 295 300
Leu Val Gln Ala Ala Tyr Gln His Val Val Gly Tyr Pro Glu Leu Met
305 310 315 320
Gly Met Met Asp Asp Ile Pro Leu Thr Asp Ile Phe Glu Asp Ala Ile
325 330 335
Ala Gly Leu Leu Ala Pro Lys Ala Met Lys Leu Leu Lys Lys Leu Leu
340 345 350
Lys Leu Gln Lys Lys Gly Ile
355
<210> 27
<211> 359
<212> PRT
<213> Artificial Sequence
<220>
<223> lipase M37 - NRC
<400> 27
Met Ser Tyr Thr Lys Glu Gln Leu Met Leu Ala Phe Ser Tyr Met Ser
1 5 10 15
Tyr Tyr Gly Ile Thr His Thr Gly Ser Ala Lys Lys Asn Ala Glu Leu
20 25 30
Ile Leu Lys Lys Met Lys Glu Ala Leu Lys Thr Trp Lys Pro Phe Gln
35 40 45
Glu Asp Asp Trp Glu Val Val Trp Gly Pro Ala Val Tyr Thr Met Pro
50 55 60
Phe Thr Ile Phe Asn Asp Ala Met Met Tyr Val Ile Gln Lys Lys Gly
65 70 75 80
Ala Glu Gly Glu Tyr Val Ile Ala Ile Arg Gly Thr Asn Pro Val Ser
85 90 95
Ile Ser Asp Trp Leu Phe Asn Asp Phe Met Val Ser Ala Met Lys Lys
100 105 110
Trp Pro Tyr Ala Ser Val Glu Gly Arg Ile Leu Lys Ile Ser Glu Ser
115 120 125
Thr Ser Tyr Gly Leu Lys Thr Leu Gln Lys Leu Lys Pro Lys Ser His
130 135 140
Ile Pro Gly Glu Asn Lys Thr Ile Leu Gln Phe Leu Asn Glu Lys Ile
145 150 155 160
Gly Pro Glu Gly Lys Ala Lys Ile Cys Val Thr Gly His Ser Lys Gly
165 170 175
Gly Ala Leu Ser Ser Thr Leu Ala Leu Trp Leu Lys Asp Ile Gln Gly
180 185 190
Val Lys Leu Ser Gln Asn Ile Asp Ile Ser Thr Ile Pro Phe Ala Gly
195 200 205
Pro Thr Ala Gly Asn Ala Asp Phe Ala Asp Tyr Phe Asp Asp Cys Leu
210 215 220
Gly Asp Gln Cys Thr Arg Ile Ala Asn Ser Leu Asp Ile Val Pro Tyr
225 230 235 240
Ala Trp Asn Thr Asn Ser Leu Lys Lys Leu Lys Ser Ile Tyr Ile Ser
245 250 255
Glu Gln Ala Ser Val Lys Pro Leu Leu Tyr Gln Arg Ala Leu Ile Arg
260 265 270
Ala Met Ile Ala Glu Thr Lys Gly Lys Lys Tyr Lys Gln Ile Lys Ala
275 280 285
Glu Thr Pro Pro Leu Glu Gly Asn Ile Asn Pro Ile Leu Ile Glu Tyr
290 295 300
Leu Val Gln Ala Ala Tyr Gln His Val Val Gly Tyr Pro Glu Leu Met
305 310 315 320
Gly Met Met Asp Asp Ile Pro Leu Thr Asp Ile Phe Glu Asp Ala Ile
325 330 335
Ala Gly Leu Leu Ala Pro Lys Ala Met Arg Leu Leu Arg Arg Leu Leu
340 345 350
Arg Leu Gln Lys Lys Gly Ile
355
<210> 28
<211> 518
<212> PRT
<213> Artificial Sequence
<220>
<223> Buforin IIb - lipase TliA - Buforin IIb
<400> 28
Arg Ala Gly Leu Gln Phe Pro Val Gly Arg Leu Leu Arg Arg Leu Leu
1 5 10 15
Arg Arg Leu Leu Arg Met Gly Val Phe Asp Tyr Lys Asn Leu Gly Thr
20 25 30
Glu Ala Ser Lys Thr Leu Phe Ala Asp Ala Thr Ala Ile Thr Leu Tyr
35 40 45
Thr Tyr His Asn Leu Asp Asn Gly Phe Ala Val Gly Tyr Gln Gln His
50 55 60
Gly Leu Gly Leu Gly Leu Pro Ala Thr Leu Val Gly Ala Leu Leu Gly
65 70 75 80
Ser Thr Asp Ser Gln Gly Val Ile Pro Gly Ile Pro Trp Asn Pro Asp
85 90 95
Ser Glu Lys Ala Ala Leu Asp Ala Val His Ala Ala Gly Trp Thr Pro
100 105 110
Ile Ser Ala Ser Ala Leu Gly Tyr Gly Gly Lys Val Asp Ala Arg Gly
115 120 125
Thr Phe Phe Gly Glu Lys Ala Gly Tyr Thr Thr Ala Gln Ala Glu Val
130 135 140
Leu Gly Lys Tyr Asp Asp Ala Gly Lys Leu Leu Glu Ile Gly Ile Gly
145 150 155 160
Phe Arg Gly Thr Ser Gly Pro Arg Glu Ser Leu Ile Thr Asp Ser Ile
165 170 175
Gly Asp Leu Val Ser Asp Leu Leu Ala Ala Leu Gly Pro Lys Asp Tyr
180 185 190
Ala Lys Asn Tyr Ala Gly Glu Ala Phe Gly Gly Leu Leu Lys Thr Val
195 200 205
Ala Asp Tyr Ala Gly Ala His Gly Leu Ser Gly Lys Asp Val Leu Val
210 215 220
Ser Gly His Ser Leu Gly Gly Leu Ala Val Asn Ser Met Ala Asp Leu
225 230 235 240
Ser Thr Ser Lys Trp Ala Gly Phe Tyr Lys Asp Ala Asn Tyr Leu Ala
245 250 255
Tyr Ala Ser Pro Thr Gln Ser Ala Gly Asp Lys Val Leu Asn Ile Gly
260 265 270
Tyr Glu Asn Asp Pro Val Phe Arg Ala Leu Asp Gly Ser Thr Phe Asn
275 280 285
Leu Ser Ser Leu Gly Val His Asp Lys Ala His Glu Ser Thr Thr Asp
290 295 300
Asn Ile Val Ser Phe Asn Asp His Tyr Ala Ser Thr Leu Trp Asn Val
305 310 315 320
Leu Pro Phe Ser Ile Ala Asn Leu Ser Thr Trp Val Ser His Leu Pro
325 330 335
Ser Ala Tyr Gly Asp Gly Met Thr Arg Val Leu Glu Ser Gly Phe Tyr
340 345 350
Glu Gln Met Thr Arg Asp Ser Thr Ile Ile Val Ala Asn Leu Ser Asp
355 360 365
Pro Ala Arg Ala Asn Thr Trp Val Gln Asp Leu Asn Arg Asn Ala Glu
370 375 380
Pro His Thr Gly Asn Thr Phe Ile Ile Gly Ser Asp Gly Asn Asp Leu
385 390 395 400
Ile Gln Gly Gly Lys Gly Ala Asp Phe Ile Glu Gly Gly Lys Gly Asn
405 410 415
Asp Thr Ile Arg Asp Asn Ser Gly His Asn Thr Phe Leu Phe Ser Gly
420 425 430
His Phe Gly Gln Asp Arg Ile Ile Gly Tyr Gln Pro Thr Asp Arg Leu
435 440 445
Val Phe Gln Gly Ala Asp Gly Ser Thr Asp Leu Arg Asp His Ala Lys
450 455 460
Ala Val Gly Ala Asp Thr Val Leu Ser Phe Gly Ala Asp Ser Val Thr
465 470 475 480
Leu Val Gly Val Gly Leu Gly Gly Leu Trp Ser Glu Gly Val Leu Ile
485 490 495
Ser Arg Ala Gly Leu Gln Phe Pro Val Gly Arg Leu Leu Arg Arg Leu
500 505 510
Leu Arg Arg Leu Leu Arg
515
<210> 29
<211> 494
<212> PRT
<213> Artificial Sequence
<220>
<223> B0 - lipase TliA -B0
<400> 29
Arg Ala Gly Leu Gln Phe Pro Val Gly Met Gly Val Phe Asp Tyr Lys
1 5 10 15
Asn Leu Gly Thr Glu Ala Ser Lys Thr Leu Phe Ala Asp Ala Thr Ala
20 25 30
Ile Thr Leu Tyr Thr Tyr His Asn Leu Asp Asn Gly Phe Ala Val Gly
35 40 45
Tyr Gln Gln His Gly Leu Gly Leu Gly Leu Pro Ala Thr Leu Val Gly
50 55 60
Ala Leu Leu Gly Ser Thr Asp Ser Gln Gly Val Ile Pro Gly Ile Pro
65 70 75 80
Trp Asn Pro Asp Ser Glu Lys Ala Ala Leu Asp Ala Val His Ala Ala
85 90 95
Gly Trp Thr Pro Ile Ser Ala Ser Ala Leu Gly Tyr Gly Gly Lys Val
100 105 110
Asp Ala Arg Gly Thr Phe Phe Gly Glu Lys Ala Gly Tyr Thr Thr Ala
115 120 125
Gln Ala Glu Val Leu Gly Lys Tyr Asp Asp Ala Gly Lys Leu Leu Glu
130 135 140
Ile Gly Ile Gly Phe Arg Gly Thr Ser Gly Pro Arg Glu Ser Leu Ile
145 150 155 160
Thr Asp Ser Ile Gly Asp Leu Val Ser Asp Leu Leu Ala Ala Leu Gly
165 170 175
Pro Lys Asp Tyr Ala Lys Asn Tyr Ala Gly Glu Ala Phe Gly Gly Leu
180 185 190
Leu Lys Thr Val Ala Asp Tyr Ala Gly Ala His Gly Leu Ser Gly Lys
195 200 205
Asp Val Leu Val Ser Gly His Ser Leu Gly Gly Leu Ala Val Asn Ser
210 215 220
Met Ala Asp Leu Ser Thr Ser Lys Trp Ala Gly Phe Tyr Lys Asp Ala
225 230 235 240
Asn Tyr Leu Ala Tyr Ala Ser Pro Thr Gln Ser Ala Gly Asp Lys Val
245 250 255
Leu Asn Ile Gly Tyr Glu Asn Asp Pro Val Phe Arg Ala Leu Asp Gly
260 265 270
Ser Thr Phe Asn Leu Ser Ser Leu Gly Val His Asp Lys Ala His Glu
275 280 285
Ser Thr Thr Asp Asn Ile Val Ser Phe Asn Asp His Tyr Ala Ser Thr
290 295 300
Leu Trp Asn Val Leu Pro Phe Ser Ile Ala Asn Leu Ser Thr Trp Val
305 310 315 320
Ser His Leu Pro Ser Ala Tyr Gly Asp Gly Met Thr Arg Val Leu Glu
325 330 335
Ser Gly Phe Tyr Glu Gln Met Thr Arg Asp Ser Thr Ile Ile Val Ala
340 345 350
Asn Leu Ser Asp Pro Ala Arg Ala Asn Thr Trp Val Gln Asp Leu Asn
355 360 365
Arg Asn Ala Glu Pro His Thr Gly Asn Thr Phe Ile Ile Gly Ser Asp
370 375 380
Gly Asn Asp Leu Ile Gln Gly Gly Lys Gly Ala Asp Phe Ile Glu Gly
385 390 395 400
Gly Lys Gly Asn Asp Thr Ile Arg Asp Asn Ser Gly His Asn Thr Phe
405 410 415
Leu Phe Ser Gly His Phe Gly Gln Asp Arg Ile Ile Gly Tyr Gln Pro
420 425 430
Thr Asp Arg Leu Val Phe Gln Gly Ala Asp Gly Ser Thr Asp Leu Arg
435 440 445
Asp His Ala Lys Ala Val Gly Ala Asp Thr Val Leu Ser Phe Gly Ala
450 455 460
Asp Ser Val Thr Leu Val Gly Val Gly Leu Gly Gly Leu Trp Ser Glu
465 470 475 480
Gly Val Leu Ile Ser Arg Ala Gly Leu Gln Phe Pro Val Gly
485 490
<210> 30
<211> 514
<212> PRT
<213> Artificial Sequence
<220>
<223> Paracin I - lipase TliA - Paracin I
<400> 30
Lys Gly Arg Gly Lys Gln Gly Gly Lys Val Arg Ala Lys Ala Lys Thr
1 5 10 15
Arg Ser Ser Met Gly Val Phe Asp Tyr Lys Asn Leu Gly Thr Glu Ala
20 25 30
Ser Lys Thr Leu Phe Ala Asp Ala Thr Ala Ile Thr Leu Tyr Thr Tyr
35 40 45
His Asn Leu Asp Asn Gly Phe Ala Val Gly Tyr Gln Gln His Gly Leu
50 55 60
Gly Leu Gly Leu Pro Ala Thr Leu Val Gly Ala Leu Leu Gly Ser Thr
65 70 75 80
Asp Ser Gln Gly Val Ile Pro Gly Ile Pro Trp Asn Pro Asp Ser Glu
85 90 95
Lys Ala Ala Leu Asp Ala Val His Ala Ala Gly Trp Thr Pro Ile Ser
100 105 110
Ala Ser Ala Leu Gly Tyr Gly Gly Lys Val Asp Ala Arg Gly Thr Phe
115 120 125
Phe Gly Glu Lys Ala Gly Tyr Thr Thr Ala Gln Ala Glu Val Leu Gly
130 135 140
Lys Tyr Asp Asp Ala Gly Lys Leu Leu Glu Ile Gly Ile Gly Phe Arg
145 150 155 160
Gly Thr Ser Gly Pro Arg Glu Ser Leu Ile Thr Asp Ser Ile Gly Asp
165 170 175
Leu Val Ser Asp Leu Leu Ala Ala Leu Gly Pro Lys Asp Tyr Ala Lys
180 185 190
Asn Tyr Ala Gly Glu Ala Phe Gly Gly Leu Leu Lys Thr Val Ala Asp
195 200 205
Tyr Ala Gly Ala His Gly Leu Ser Gly Lys Asp Val Leu Val Ser Gly
210 215 220
His Ser Leu Gly Gly Leu Ala Val Asn Ser Met Ala Asp Leu Ser Thr
225 230 235 240
Ser Lys Trp Ala Gly Phe Tyr Lys Asp Ala Asn Tyr Leu Ala Tyr Ala
245 250 255
Ser Pro Thr Gln Ser Ala Gly Asp Lys Val Leu Asn Ile Gly Tyr Glu
260 265 270
Asn Asp Pro Val Phe Arg Ala Leu Asp Gly Ser Thr Phe Asn Leu Ser
275 280 285
Ser Leu Gly Val His Asp Lys Ala His Glu Ser Thr Thr Asp Asn Ile
290 295 300
Val Ser Phe Asn Asp His Tyr Ala Ser Thr Leu Trp Asn Val Leu Pro
305 310 315 320
Phe Ser Ile Ala Asn Leu Ser Thr Trp Val Ser His Leu Pro Ser Ala
325 330 335
Tyr Gly Asp Gly Met Thr Arg Val Leu Glu Ser Gly Phe Tyr Glu Gln
340 345 350
Met Thr Arg Asp Ser Thr Ile Ile Val Ala Asn Leu Ser Asp Pro Ala
355 360 365
Arg Ala Asn Thr Trp Val Gln Asp Leu Asn Arg Asn Ala Glu Pro His
370 375 380
Thr Gly Asn Thr Phe Ile Ile Gly Ser Asp Gly Asn Asp Leu Ile Gln
385 390 395 400
Gly Gly Lys Gly Ala Asp Phe Ile Glu Gly Gly Lys Gly Asn Asp Thr
405 410 415
Ile Arg Asp Asn Ser Gly His Asn Thr Phe Leu Phe Ser Gly His Phe
420 425 430
Gly Gln Asp Arg Ile Ile Gly Tyr Gln Pro Thr Asp Arg Leu Val Phe
435 440 445
Gln Gly Ala Asp Gly Ser Thr Asp Leu Arg Asp His Ala Lys Ala Val
450 455 460
Gly Ala Asp Thr Val Leu Ser Phe Gly Ala Asp Ser Val Thr Leu Val
465 470 475 480
Gly Val Gly Leu Gly Gly Leu Trp Ser Glu Gly Val Leu Ile Ser Lys
485 490 495
Gly Arg Gly Lys Gln Gly Gly Lys Val Arg Ala Lys Ala Lys Thr Arg
500 505 510
Ser Ser
<210> 31
<211> 514
<212> PRT
<213> Artificial Sequence
<220>
<223> NKC - lipase TliA - NKC
<400> 31
Ala Pro Lys Ala Met Lys Leu Leu Lys Lys Leu Leu Lys Leu Gln Lys
1 5 10 15
Lys Gly Ile Met Gly Val Phe Asp Tyr Lys Asn Leu Gly Thr Glu Ala
20 25 30
Ser Lys Thr Leu Phe Ala Asp Ala Thr Ala Ile Thr Leu Tyr Thr Tyr
35 40 45
His Asn Leu Asp Asn Gly Phe Ala Val Gly Tyr Gln Gln His Gly Leu
50 55 60
Gly Leu Gly Leu Pro Ala Thr Leu Val Gly Ala Leu Leu Gly Ser Thr
65 70 75 80
Asp Ser Gln Gly Val Ile Pro Gly Ile Pro Trp Asn Pro Asp Ser Glu
85 90 95
Lys Ala Ala Leu Asp Ala Val His Ala Ala Gly Trp Thr Pro Ile Ser
100 105 110
Ala Ser Ala Leu Gly Tyr Gly Gly Lys Val Asp Ala Arg Gly Thr Phe
115 120 125
Phe Gly Glu Lys Ala Gly Tyr Thr Thr Ala Gln Ala Glu Val Leu Gly
130 135 140
Lys Tyr Asp Asp Ala Gly Lys Leu Leu Glu Ile Gly Ile Gly Phe Arg
145 150 155 160
Gly Thr Ser Gly Pro Arg Glu Ser Leu Ile Thr Asp Ser Ile Gly Asp
165 170 175
Leu Val Ser Asp Leu Leu Ala Ala Leu Gly Pro Lys Asp Tyr Ala Lys
180 185 190
Asn Tyr Ala Gly Glu Ala Phe Gly Gly Leu Leu Lys Thr Val Ala Asp
195 200 205
Tyr Ala Gly Ala His Gly Leu Ser Gly Lys Asp Val Leu Val Ser Gly
210 215 220
His Ser Leu Gly Gly Leu Ala Val Asn Ser Met Ala Asp Leu Ser Thr
225 230 235 240
Ser Lys Trp Ala Gly Phe Tyr Lys Asp Ala Asn Tyr Leu Ala Tyr Ala
245 250 255
Ser Pro Thr Gln Ser Ala Gly Asp Lys Val Leu Asn Ile Gly Tyr Glu
260 265 270
Asn Asp Pro Val Phe Arg Ala Leu Asp Gly Ser Thr Phe Asn Leu Ser
275 280 285
Ser Leu Gly Val His Asp Lys Ala His Glu Ser Thr Thr Asp Asn Ile
290 295 300
Val Ser Phe Asn Asp His Tyr Ala Ser Thr Leu Trp Asn Val Leu Pro
305 310 315 320
Phe Ser Ile Ala Asn Leu Ser Thr Trp Val Ser His Leu Pro Ser Ala
325 330 335
Tyr Gly Asp Gly Met Thr Arg Val Leu Glu Ser Gly Phe Tyr Glu Gln
340 345 350
Met Thr Arg Asp Ser Thr Ile Ile Val Ala Asn Leu Ser Asp Pro Ala
355 360 365
Arg Ala Asn Thr Trp Val Gln Asp Leu Asn Arg Asn Ala Glu Pro His
370 375 380
Thr Gly Asn Thr Phe Ile Ile Gly Ser Asp Gly Asn Asp Leu Ile Gln
385 390 395 400
Gly Gly Lys Gly Ala Asp Phe Ile Glu Gly Gly Lys Gly Asn Asp Thr
405 410 415
Ile Arg Asp Asn Ser Gly His Asn Thr Phe Leu Phe Ser Gly His Phe
420 425 430
Gly Gln Asp Arg Ile Ile Gly Tyr Gln Pro Thr Asp Arg Leu Val Phe
435 440 445
Gln Gly Ala Asp Gly Ser Thr Asp Leu Arg Asp His Ala Lys Ala Val
450 455 460
Gly Ala Asp Thr Val Leu Ser Phe Gly Ala Asp Ser Val Thr Leu Val
465 470 475 480
Gly Val Gly Leu Gly Gly Leu Trp Ser Glu Gly Val Leu Ile Ser Ala
485 490 495
Pro Lys Ala Met Lys Leu Leu Lys Lys Leu Leu Lys Leu Gln Lys Lys
500 505 510
Gly Ile
<210> 32
<211> 514
<212> PRT
<213> Artificial Sequence
<220>
<223> NRC - lipase TliA - NRC
<400> 32
Ala Pro Lys Ala Met Arg Leu Leu Arg Arg Leu Leu Arg Leu Gln Lys
1 5 10 15
Lys Gly Ile Met Gly Val Phe Asp Tyr Lys Asn Leu Gly Thr Glu Ala
20 25 30
Ser Lys Thr Leu Phe Ala Asp Ala Thr Ala Ile Thr Leu Tyr Thr Tyr
35 40 45
His Asn Leu Asp Asn Gly Phe Ala Val Gly Tyr Gln Gln His Gly Leu
50 55 60
Gly Leu Gly Leu Pro Ala Thr Leu Val Gly Ala Leu Leu Gly Ser Thr
65 70 75 80
Asp Ser Gln Gly Val Ile Pro Gly Ile Pro Trp Asn Pro Asp Ser Glu
85 90 95
Lys Ala Ala Leu Asp Ala Val His Ala Ala Gly Trp Thr Pro Ile Ser
100 105 110
Ala Ser Ala Leu Gly Tyr Gly Gly Lys Val Asp Ala Arg Gly Thr Phe
115 120 125
Phe Gly Glu Lys Ala Gly Tyr Thr Thr Ala Gln Ala Glu Val Leu Gly
130 135 140
Lys Tyr Asp Asp Ala Gly Lys Leu Leu Glu Ile Gly Ile Gly Phe Arg
145 150 155 160
Gly Thr Ser Gly Pro Arg Glu Ser Leu Ile Thr Asp Ser Ile Gly Asp
165 170 175
Leu Val Ser Asp Leu Leu Ala Ala Leu Gly Pro Lys Asp Tyr Ala Lys
180 185 190
Asn Tyr Ala Gly Glu Ala Phe Gly Gly Leu Leu Lys Thr Val Ala Asp
195 200 205
Tyr Ala Gly Ala His Gly Leu Ser Gly Lys Asp Val Leu Val Ser Gly
210 215 220
His Ser Leu Gly Gly Leu Ala Val Asn Ser Met Ala Asp Leu Ser Thr
225 230 235 240
Ser Lys Trp Ala Gly Phe Tyr Lys Asp Ala Asn Tyr Leu Ala Tyr Ala
245 250 255
Ser Pro Thr Gln Ser Ala Gly Asp Lys Val Leu Asn Ile Gly Tyr Glu
260 265 270
Asn Asp Pro Val Phe Arg Ala Leu Asp Gly Ser Thr Phe Asn Leu Ser
275 280 285
Ser Leu Gly Val His Asp Lys Ala His Glu Ser Thr Thr Asp Asn Ile
290 295 300
Val Ser Phe Asn Asp His Tyr Ala Ser Thr Leu Trp Asn Val Leu Pro
305 310 315 320
Phe Ser Ile Ala Asn Leu Ser Thr Trp Val Ser His Leu Pro Ser Ala
325 330 335
Tyr Gly Asp Gly Met Thr Arg Val Leu Glu Ser Gly Phe Tyr Glu Gln
340 345 350
Met Thr Arg Asp Ser Thr Ile Ile Val Ala Asn Leu Ser Asp Pro Ala
355 360 365
Arg Ala Asn Thr Trp Val Gln Asp Leu Asn Arg Asn Ala Glu Pro His
370 375 380
Thr Gly Asn Thr Phe Ile Ile Gly Ser Asp Gly Asn Asp Leu Ile Gln
385 390 395 400
Gly Gly Lys Gly Ala Asp Phe Ile Glu Gly Gly Lys Gly Asn Asp Thr
405 410 415
Ile Arg Asp Asn Ser Gly His Asn Thr Phe Leu Phe Ser Gly His Phe
420 425 430
Gly Gln Asp Arg Ile Ile Gly Tyr Gln Pro Thr Asp Arg Leu Val Phe
435 440 445
Gln Gly Ala Asp Gly Ser Thr Asp Leu Arg Asp His Ala Lys Ala Val
450 455 460
Gly Ala Asp Thr Val Leu Ser Phe Gly Ala Asp Ser Val Thr Leu Val
465 470 475 480
Gly Val Gly Leu Gly Gly Leu Trp Ser Glu Gly Val Leu Ile Ser Ala
485 490 495
Pro Lys Ala Met Arg Leu Leu Arg Arg Leu Leu Arg Leu Gln Lys Lys
500 505 510
Gly Ile
<210> 33
<211> 382
<212> PRT
<213> Artificial Sequence
<220>
<223> Buforin IIb - lipase M37 - Buforin IIb
<400> 33
Arg Ala Gly Leu Gln Phe Pro Val Gly Arg Leu Leu Arg Arg Leu Leu
1 5 10 15
Arg Arg Leu Leu Arg Met Ser Tyr Thr Lys Glu Gln Leu Met Leu Ala
20 25 30
Phe Ser Tyr Met Ser Tyr Tyr Gly Ile Thr His Thr Gly Ser Ala Lys
35 40 45
Lys Asn Ala Glu Leu Ile Leu Lys Lys Met Lys Glu Ala Leu Lys Thr
50 55 60
Trp Lys Pro Phe Gln Glu Asp Asp Trp Glu Val Val Trp Gly Pro Ala
65 70 75 80
Val Tyr Thr Met Pro Phe Thr Ile Phe Asn Asp Ala Met Met Tyr Val
85 90 95
Ile Gln Lys Lys Gly Ala Glu Gly Glu Tyr Val Ile Ala Ile Arg Gly
100 105 110
Thr Asn Pro Val Ser Ile Ser Asp Trp Leu Phe Asn Asp Phe Met Val
115 120 125
Ser Ala Met Lys Lys Trp Pro Tyr Ala Ser Val Glu Gly Arg Ile Leu
130 135 140
Lys Ile Ser Glu Ser Thr Ser Tyr Gly Leu Lys Thr Leu Gln Lys Leu
145 150 155 160
Lys Pro Lys Ser His Ile Pro Gly Glu Asn Lys Thr Ile Leu Gln Phe
165 170 175
Leu Asn Glu Lys Ile Gly Pro Glu Gly Lys Ala Lys Ile Cys Val Thr
180 185 190
Gly His Ser Lys Gly Gly Ala Leu Ser Ser Thr Leu Ala Leu Trp Leu
195 200 205
Lys Asp Ile Gln Gly Val Lys Leu Ser Gln Asn Ile Asp Ile Ser Thr
210 215 220
Ile Pro Phe Ala Gly Pro Thr Ala Gly Asn Ala Asp Phe Ala Asp Tyr
225 230 235 240
Phe Asp Asp Cys Leu Gly Asp Gln Cys Thr Arg Ile Ala Asn Ser Leu
245 250 255
Asp Ile Val Pro Tyr Ala Trp Asn Thr Asn Ser Leu Lys Lys Leu Lys
260 265 270
Ser Ile Tyr Ile Ser Glu Gln Ala Ser Val Lys Pro Leu Leu Tyr Gln
275 280 285
Arg Ala Leu Ile Arg Ala Met Ile Ala Glu Thr Lys Gly Lys Lys Tyr
290 295 300
Lys Gln Ile Lys Ala Glu Thr Pro Pro Leu Glu Gly Asn Ile Asn Pro
305 310 315 320
Ile Leu Ile Glu Tyr Leu Val Gln Ala Ala Tyr Gln His Val Val Gly
325 330 335
Tyr Pro Glu Leu Met Gly Met Met Asp Asp Ile Pro Leu Thr Asp Ile
340 345 350
Phe Glu Asp Ala Ile Ala Gly Leu Leu Arg Ala Gly Leu Gln Phe Pro
355 360 365
Val Gly Arg Leu Leu Arg Arg Leu Leu Arg Arg Leu Leu Arg
370 375 380
<210> 34
<211> 358
<212> PRT
<213> Artificial Sequence
<220>
<223> B0 - lipase M37 - B0
<400> 34
Arg Ala Gly Leu Gln Phe Pro Val Gly Met Ser Tyr Thr Lys Glu Gln
1 5 10 15
Leu Met Leu Ala Phe Ser Tyr Met Ser Tyr Tyr Gly Ile Thr His Thr
20 25 30
Gly Ser Ala Lys Lys Asn Ala Glu Leu Ile Leu Lys Lys Met Lys Glu
35 40 45
Ala Leu Lys Thr Trp Lys Pro Phe Gln Glu Asp Asp Trp Glu Val Val
50 55 60
Trp Gly Pro Ala Val Tyr Thr Met Pro Phe Thr Ile Phe Asn Asp Ala
65 70 75 80
Met Met Tyr Val Ile Gln Lys Lys Gly Ala Glu Gly Glu Tyr Val Ile
85 90 95
Ala Ile Arg Gly Thr Asn Pro Val Ser Ile Ser Asp Trp Leu Phe Asn
100 105 110
Asp Phe Met Val Ser Ala Met Lys Lys Trp Pro Tyr Ala Ser Val Glu
115 120 125
Gly Arg Ile Leu Lys Ile Ser Glu Ser Thr Ser Tyr Gly Leu Lys Thr
130 135 140
Leu Gln Lys Leu Lys Pro Lys Ser His Ile Pro Gly Glu Asn Lys Thr
145 150 155 160
Ile Leu Gln Phe Leu Asn Glu Lys Ile Gly Pro Glu Gly Lys Ala Lys
165 170 175
Ile Cys Val Thr Gly His Ser Lys Gly Gly Ala Leu Ser Ser Thr Leu
180 185 190
Ala Leu Trp Leu Lys Asp Ile Gln Gly Val Lys Leu Ser Gln Asn Ile
195 200 205
Asp Ile Ser Thr Ile Pro Phe Ala Gly Pro Thr Ala Gly Asn Ala Asp
210 215 220
Phe Ala Asp Tyr Phe Asp Asp Cys Leu Gly Asp Gln Cys Thr Arg Ile
225 230 235 240
Ala Asn Ser Leu Asp Ile Val Pro Tyr Ala Trp Asn Thr Asn Ser Leu
245 250 255
Lys Lys Leu Lys Ser Ile Tyr Ile Ser Glu Gln Ala Ser Val Lys Pro
260 265 270
Leu Leu Tyr Gln Arg Ala Leu Ile Arg Ala Met Ile Ala Glu Thr Lys
275 280 285
Gly Lys Lys Tyr Lys Gln Ile Lys Ala Glu Thr Pro Pro Leu Glu Gly
290 295 300
Asn Ile Asn Pro Ile Leu Ile Glu Tyr Leu Val Gln Ala Ala Tyr Gln
305 310 315 320
His Val Val Gly Tyr Pro Glu Leu Met Gly Met Met Asp Asp Ile Pro
325 330 335
Leu Thr Asp Ile Phe Glu Asp Ala Ile Ala Gly Leu Leu Arg Ala Gly
340 345 350
Leu Gln Phe Pro Val Gly
355
<210> 35
<211> 378
<212> PRT
<213> Artificial Sequence
<220>
<223> Paracin I - lipase M37 - Paracin I
<400> 35
Lys Gly Arg Gly Lys Gln Gly Gly Lys Val Arg Ala Lys Ala Lys Thr
1 5 10 15
Arg Ser Ser Met Ser Tyr Thr Lys Glu Gln Leu Met Leu Ala Phe Ser
20 25 30
Tyr Met Ser Tyr Tyr Gly Ile Thr His Thr Gly Ser Ala Lys Lys Asn
35 40 45
Ala Glu Leu Ile Leu Lys Lys Met Lys Glu Ala Leu Lys Thr Trp Lys
50 55 60
Pro Phe Gln Glu Asp Asp Trp Glu Val Val Trp Gly Pro Ala Val Tyr
65 70 75 80
Thr Met Pro Phe Thr Ile Phe Asn Asp Ala Met Met Tyr Val Ile Gln
85 90 95
Lys Lys Gly Ala Glu Gly Glu Tyr Val Ile Ala Ile Arg Gly Thr Asn
100 105 110
Pro Val Ser Ile Ser Asp Trp Leu Phe Asn Asp Phe Met Val Ser Ala
115 120 125
Met Lys Lys Trp Pro Tyr Ala Ser Val Glu Gly Arg Ile Leu Lys Ile
130 135 140
Ser Glu Ser Thr Ser Tyr Gly Leu Lys Thr Leu Gln Lys Leu Lys Pro
145 150 155 160
Lys Ser His Ile Pro Gly Glu Asn Lys Thr Ile Leu Gln Phe Leu Asn
165 170 175
Glu Lys Ile Gly Pro Glu Gly Lys Ala Lys Ile Cys Val Thr Gly His
180 185 190
Ser Lys Gly Gly Ala Leu Ser Ser Thr Leu Ala Leu Trp Leu Lys Asp
195 200 205
Ile Gln Gly Val Lys Leu Ser Gln Asn Ile Asp Ile Ser Thr Ile Pro
210 215 220
Phe Ala Gly Pro Thr Ala Gly Asn Ala Asp Phe Ala Asp Tyr Phe Asp
225 230 235 240
Asp Cys Leu Gly Asp Gln Cys Thr Arg Ile Ala Asn Ser Leu Asp Ile
245 250 255
Val Pro Tyr Ala Trp Asn Thr Asn Ser Leu Lys Lys Leu Lys Ser Ile
260 265 270
Tyr Ile Ser Glu Gln Ala Ser Val Lys Pro Leu Leu Tyr Gln Arg Ala
275 280 285
Leu Ile Arg Ala Met Ile Ala Glu Thr Lys Gly Lys Lys Tyr Lys Gln
290 295 300
Ile Lys Ala Glu Thr Pro Pro Leu Glu Gly Asn Ile Asn Pro Ile Leu
305 310 315 320
Ile Glu Tyr Leu Val Gln Ala Ala Tyr Gln His Val Val Gly Tyr Pro
325 330 335
Glu Leu Met Gly Met Met Asp Asp Ile Pro Leu Thr Asp Ile Phe Glu
340 345 350
Asp Ala Ile Ala Gly Leu Leu Lys Gly Arg Gly Lys Gln Gly Gly Lys
355 360 365
Val Arg Ala Lys Ala Lys Thr Arg Ser Ser
370 375
<210> 36
<211> 378
<212> PRT
<213> Artificial Sequence
<220>
<223> NKC - lipase M37 - NKC
<400> 36
Ala Pro Lys Ala Met Lys Leu Leu Lys Lys Leu Leu Lys Leu Gln Lys
1 5 10 15
Lys Gly Ile Met Ser Tyr Thr Lys Glu Gln Leu Met Leu Ala Phe Ser
20 25 30
Tyr Met Ser Tyr Tyr Gly Ile Thr His Thr Gly Ser Ala Lys Lys Asn
35 40 45
Ala Glu Leu Ile Leu Lys Lys Met Lys Glu Ala Leu Lys Thr Trp Lys
50 55 60
Pro Phe Gln Glu Asp Asp Trp Glu Val Val Trp Gly Pro Ala Val Tyr
65 70 75 80
Thr Met Pro Phe Thr Ile Phe Asn Asp Ala Met Met Tyr Val Ile Gln
85 90 95
Lys Lys Gly Ala Glu Gly Glu Tyr Val Ile Ala Ile Arg Gly Thr Asn
100 105 110
Pro Val Ser Ile Ser Asp Trp Leu Phe Asn Asp Phe Met Val Ser Ala
115 120 125
Met Lys Lys Trp Pro Tyr Ala Ser Val Glu Gly Arg Ile Leu Lys Ile
130 135 140
Ser Glu Ser Thr Ser Tyr Gly Leu Lys Thr Leu Gln Lys Leu Lys Pro
145 150 155 160
Lys Ser His Ile Pro Gly Glu Asn Lys Thr Ile Leu Gln Phe Leu Asn
165 170 175
Glu Lys Ile Gly Pro Glu Gly Lys Ala Lys Ile Cys Val Thr Gly His
180 185 190
Ser Lys Gly Gly Ala Leu Ser Ser Thr Leu Ala Leu Trp Leu Lys Asp
195 200 205
Ile Gln Gly Val Lys Leu Ser Gln Asn Ile Asp Ile Ser Thr Ile Pro
210 215 220
Phe Ala Gly Pro Thr Ala Gly Asn Ala Asp Phe Ala Asp Tyr Phe Asp
225 230 235 240
Asp Cys Leu Gly Asp Gln Cys Thr Arg Ile Ala Asn Ser Leu Asp Ile
245 250 255
Val Pro Tyr Ala Trp Asn Thr Asn Ser Leu Lys Lys Leu Lys Ser Ile
260 265 270
Tyr Ile Ser Glu Gln Ala Ser Val Lys Pro Leu Leu Tyr Gln Arg Ala
275 280 285
Leu Ile Arg Ala Met Ile Ala Glu Thr Lys Gly Lys Lys Tyr Lys Gln
290 295 300
Ile Lys Ala Glu Thr Pro Pro Leu Glu Gly Asn Ile Asn Pro Ile Leu
305 310 315 320
Ile Glu Tyr Leu Val Gln Ala Ala Tyr Gln His Val Val Gly Tyr Pro
325 330 335
Glu Leu Met Gly Met Met Asp Asp Ile Pro Leu Thr Asp Ile Phe Glu
340 345 350
Asp Ala Ile Ala Gly Leu Leu Ala Pro Lys Ala Met Lys Leu Leu Lys
355 360 365
Lys Leu Leu Lys Leu Gln Lys Lys Gly Ile
370 375
<210> 37
<211> 378
<212> PRT
<213> Artificial Sequence
<220>
<223> NRC - lipase M37 - NRC
<400> 37
Ala Pro Lys Ala Met Arg Leu Leu Arg Arg Leu Leu Arg Leu Gln Lys
1 5 10 15
Lys Gly Ile Met Ser Tyr Thr Lys Glu Gln Leu Met Leu Ala Phe Ser
20 25 30
Tyr Met Ser Tyr Tyr Gly Ile Thr His Thr Gly Ser Ala Lys Lys Asn
35 40 45
Ala Glu Leu Ile Leu Lys Lys Met Lys Glu Ala Leu Lys Thr Trp Lys
50 55 60
Pro Phe Gln Glu Asp Asp Trp Glu Val Val Trp Gly Pro Ala Val Tyr
65 70 75 80
Thr Met Pro Phe Thr Ile Phe Asn Asp Ala Met Met Tyr Val Ile Gln
85 90 95
Lys Lys Gly Ala Glu Gly Glu Tyr Val Ile Ala Ile Arg Gly Thr Asn
100 105 110
Pro Val Ser Ile Ser Asp Trp Leu Phe Asn Asp Phe Met Val Ser Ala
115 120 125
Met Lys Lys Trp Pro Tyr Ala Ser Val Glu Gly Arg Ile Leu Lys Ile
130 135 140
Ser Glu Ser Thr Ser Tyr Gly Leu Lys Thr Leu Gln Lys Leu Lys Pro
145 150 155 160
Lys Ser His Ile Pro Gly Glu Asn Lys Thr Ile Leu Gln Phe Leu Asn
165 170 175
Glu Lys Ile Gly Pro Glu Gly Lys Ala Lys Ile Cys Val Thr Gly His
180 185 190
Ser Lys Gly Gly Ala Leu Ser Ser Thr Leu Ala Leu Trp Leu Lys Asp
195 200 205
Ile Gln Gly Val Lys Leu Ser Gln Asn Ile Asp Ile Ser Thr Ile Pro
210 215 220
Phe Ala Gly Pro Thr Ala Gly Asn Ala Asp Phe Ala Asp Tyr Phe Asp
225 230 235 240
Asp Cys Leu Gly Asp Gln Cys Thr Arg Ile Ala Asn Ser Leu Asp Ile
245 250 255
Val Pro Tyr Ala Trp Asn Thr Asn Ser Leu Lys Lys Leu Lys Ser Ile
260 265 270
Tyr Ile Ser Glu Gln Ala Ser Val Lys Pro Leu Leu Tyr Gln Arg Ala
275 280 285
Leu Ile Arg Ala Met Ile Ala Glu Thr Lys Gly Lys Lys Tyr Lys Gln
290 295 300
Ile Lys Ala Glu Thr Pro Pro Leu Glu Gly Asn Ile Asn Pro Ile Leu
305 310 315 320
Ile Glu Tyr Leu Val Gln Ala Ala Tyr Gln His Val Val Gly Tyr Pro
325 330 335
Glu Leu Met Gly Met Met Asp Asp Ile Pro Leu Thr Asp Ile Phe Glu
340 345 350
Asp Ala Ile Ala Gly Leu Leu Ala Pro Lys Ala Met Arg Leu Leu Arg
355 360 365
Arg Leu Leu Arg Leu Gln Lys Lys Gly Ile
370 375
<210> 38
<211> 63
<212> DNA
<213> Artificial Sequence
<220>
<223> Buforin IIb
<400> 38
cgtgctggtc tgcagttccc ggttggtcgt ctgctgcgtc gtctgctgcg tcgtctgctg 60
cgt 63
<210> 39
<211> 27
<212> DNA
<213> Artificial Sequence
<220>
<223> B0
<400> 39
cgtgctggtc tgcagttccc ggttggt 27
<210> 40
<211> 57
<212> DNA
<213> Artificial Sequence
<220>
<223> Paracin I
<400> 40
aaaggaagag gcaaacaggg aggcaaggtg cgtgcgaagg caaagacacg ttcatcc 57
<210> 41
<211> 57
<212> DNA
<213> Artificial Sequence
<220>
<223> NKC
<400> 41
gcgccgaaag cgatgaaact gttgaagaaa ttgctgaaat tacagaaaaa aggcatt 57
<210> 42
<211> 57
<212> DNA
<213> Artificial Sequence
<220>
<223> NRC
<400> 42
gcgccgaaag cgatgcgtct gttgcgtcgc ttgctgcgtt tacagaaaaa aggcatt 57
<210> 43
<211> 1432
<212> DNA
<213> Artificial Sequence
<220>
<223> lipase TliA
<400> 43
atgggtgtat ttgactacaa gaacctcggc accgaagcca gcaaaacctt gttcgccgat 60
gccaccgcaa tcacgttgta tacctatcac aacctggata acggcttcgc agtcggctac 120
cagcaacatg gcttggggct cggcctgccg gccacactgg tcggggcgtt gctcggcagc 180
acagactccc agggagtgat ccccggcatt ccctggaatc ctgactcgga aaaggccgcc 240
ctggacgcgg tgcacgcagc cggttggacg ccaatcagcg ccagcgcact gggctacggc 300
ggcaaggtgg atgcgcgggg cacttttttt ggcgagaagg ccggctacac cacggcccag 360
gccgaagtgc tgggcaagta cgatgacgcc ggcaaactgc tcgagatcgg catcggtttt 420
cgtggcacct cgggccctcg ggaaagcctg attaccgact ccatcggcga tctggtcagc 480
gacctgctcg ccgcgctggg ccccaaggac tatgcgaaaa actatgccgg cgaagcgttt 540
ggcggcttgc tcaagacggt ggccgactat gccggcgccc atggcctgag tggcaaggat 600
gtgctggtca gcggccacag cctgggcggc ctggcggtca acagcatggc cgacctgagc 660
accagcaaat gggcgggttt ctacaaggac gccaactacc tggcctacgc ctcgcccacc 720
cagagcgccg gcgataaggt cctgaatatc ggctacgaaa acgacccggt attccgtgcg 780
ctggacggct ccaccttcaa cctgtcgtcc ctcggcgtgc atgacaaggc ccacgagtcg 840
accaccgaca acatcgtcag cttcaacgac cactacgcct cgacgttgtg gaatgtgctg 900
ccgttttcca tcgccaacct gtcgacctgg gtgtcgcatt tgccagcgct tacggcgacg 960
gcatgacgcg tgtgctggaa tcggggttct acgagcaaat gacccgtgac tcgacgatta 1020
tcgtcgccaa cctgtccgac ccggcgcgcg ccaacacctg ggtccaggac ctcaaccgca 1080
atgccgagcc gcacacaggc aataccttca tcatcggcag cgacggcaat gacctgatcc 1140
agggcggcaa gggcgcggac ttcatcgaag gcggcaaggg caatgacacg atccgcgaca 1200
acagcgggca caacaccttt ttgttcagcg ggcattttgg ccaggatcgg attatcggct 1260
accagccgac cgacaggctg gtgttccagg gcgccgacgg cagcaccgac ctgcgcgacc 1320
acgcgaaggc cgtgggggcc gatacggtgc tgagttttgg cgccgactcg gtgaccctgg 1380
tcggggtcgg gctgggcggc ctgtggagcg agggtgtgct gatcagttga gg 1432
<210> 44
<211> 1022
<212> DNA
<213> Artificial Sequence
<220>
<223> lipase M37
<400> 44
atgtcttata caaaagaaca actcatgttg gcattcagct atatgagcta ctatggcatc 60
actcacacag gttcagcaaa aaaaaatgct gagctcatcc ttaaaaaaat gaaagaagcc 120
ttgaaaacat ggaagccttt tcaggaagat gactgggaag tcgtctgggg tcctgcggtt 180
tataccatgc ctttcacaat cttcaacgat gccatgatgt atgtcataca gaagaaaggt 240
gctgaagggg aatacgtgat agccattcgc ggcaccaatc cagtatcaat ttcagactgg 300
ctgtttaatg atttcatggt cagcgcaatg aagaagtggc cttacgcatc cgttgaaggc 360
cgcatactca aaatatccga aagtaccagc tacggactga aaaccttaca gaaattgaag 420
ccaaaatccc atatccccgg cgaaaataaa acgattctgc agttcctgaa tgaaaagata 480
ggcccagagg gtaaagcaaa aatctgtgta acaggccaca gtaaaggcgg cgccttgtct 540
tccactctgg cactgtggtt gaaggacatc caaggagtaa aactctcgca aaacatcgat 600
atctcaacga ttccgtttgc cggaccaaca gccggtaatg ctgactttgc cgattacttt 660
gatgattgtc ttggtgatca atgcacccgc attgccaact cgttagatat tgtgccttat 720
gcctggaata aaattcatta aaaaaactta aatctatata tatttctgaa caagcatcag 780
ttaaaccgct tctatatcaa cgcgctttaa tccgtgcaat gatcgcagaa actaaaggta 840
aaaaatacaa gcaaattaag gcggaaacac caccattaga aggcaacatt aatcctattc 900
ttattgaata ccttgtgcag gcagcatatc agcatgtcgt cggttaccca gaattaatgg 960
gtatgatgga tgatattcct ttaacagaca tattcgaaga tgcgatcgcg ggtttgttat 1020
aa 1022
<210> 45
<211> 1495
<212> DNA
<213> Artificial Sequence
<220>
<223> Buforin IIb - lipase TliA
<400> 45
cgtgctggtc tgcagttccc ggttggtcgt ctgctgcgtc gtctgctgcg tcgtctgctg 60
cgtatgggtg tatttgacta caagaacctc ggcaccgaag ccagcaaaac cttgttcgcc 120
gatgccaccg caatcacgtt gtatacctat cacaacctgg ataacggctt cgcagtcggc 180
taccagcaac atggcttggg gctcggcctg ccggccacac tggtcggggc gttgctcggc 240
agcacagact cccagggagt gatccccggc attccctgga atcctgactc ggaaaaggcc 300
gccctggacg cggtgcacgc agccggttgg acgccaatca gcgccagcgc actgggctac 360
ggcggcaagg tggatgcgcg gggcactttt tttggcgaga aggccggcta caccacggcc 420
caggccgaag tgctgggcaa gtacgatgac gccggcaaac tgctcgagat cggcatcggt 480
tttcgtggca cctcgggccc tcgggaaagc ctgattaccg actccatcgg cgatctggtc 540
agcgacctgc tcgccgcgct gggccccaag gactatgcga aaaactatgc cggcgaagcg 600
tttggcggct tgctcaagac ggtggccgac tatgccggcg cccatggcct gagtggcaag 660
gatgtgctgg tcagcggcca cagcctgggc ggcctggcgg tcaacagcat ggccgacctg 720
agcaccagca aatgggcggg tttctacaag gacgccaact acctggccta cgcctcgccc 780
acccagagcg ccggcgataa ggtcctgaat atcggctacg aaaacgaccc ggtattccgt 840
gcgctggacg gctccacctt caacctgtcg tccctcggcg tgcatgacaa ggcccacgag 900
tcgaccaccg acaacatcgt cagcttcaac gaccactacg cctcgacgtt gtggaatgtg 960
ctgccgtttt ccatcgccaa cctgtcgacc tgggtgtcgc atttgccagc gcttacggcg 1020
acggcatgac gcgtgtgctg gaatcggggt tctacgagca aatgacccgt gactcgacga 1080
ttatcgtcgc caacctgtcc gacccggcgc gcgccaacac ctgggtccag gacctcaacc 1140
gcaatgccga gccgcacaca ggcaatacct tcatcatcgg cagcgacggc aatgacctga 1200
tccagggcgg caagggcgcg gacttcatcg aaggcggcaa gggcaatgac acgatccgcg 1260
acaacagcgg gcacaacacc tttttgttca gcgggcattt tggccaggat cggattatcg 1320
gctaccagcc gaccgacagg ctggtgttcc agggcgccga cggcagcacc gacctgcgcg 1380
accacgcgaa ggccgtgggg gccgatacgg tgctgagttt tggcgccgac tcggtgaccc 1440
tggtcggggt cgggctgggc ggcctgtgga gcgagggtgt gctgatcagt tgagg 1495
<210> 46
<211> 1459
<212> DNA
<213> Artificial Sequence
<220>
<223> B0 - lipase TliA
<400> 46
cgtgctggtc tgcagttccc ggttggtatg ggtgtatttg actacaagaa cctcggcacc 60
gaagccagca aaaccttgtt cgccgatgcc accgcaatca cgttgtatac ctatcacaac 120
ctggataacg gcttcgcagt cggctaccag caacatggct tggggctcgg cctgccggcc 180
acactggtcg gggcgttgct cggcagcaca gactcccagg gagtgatccc cggcattccc 240
tggaatcctg actcggaaaa ggccgccctg gacgcggtgc acgcagccgg ttggacgcca 300
atcagcgcca gcgcactggg ctacggcggc aaggtggatg cgcggggcac tttttttggc 360
gagaaggccg gctacaccac ggcccaggcc gaagtgctgg gcaagtacga tgacgccggc 420
aaactgctcg agatcggcat cggttttcgt ggcacctcgg gccctcggga aagcctgatt 480
accgactcca tcggcgatct ggtcagcgac ctgctcgccg cgctgggccc caaggactat 540
gcgaaaaact atgccggcga agcgtttggc ggcttgctca agacggtggc cgactatgcc 600
ggcgcccatg gcctgagtgg caaggatgtg ctggtcagcg gccacagcct gggcggcctg 660
gcggtcaaca gcatggccga cctgagcacc agcaaatggg cgggtttcta caaggacgcc 720
aactacctgg cctacgcctc gcccacccag agcgccggcg ataaggtcct gaatatcggc 780
tacgaaaacg acccggtatt ccgtgcgctg gacggctcca ccttcaacct gtcgtccctc 840
ggcgtgcatg acaaggccca cgagtcgacc accgacaaca tcgtcagctt caacgaccac 900
tacgcctcga cgttgtggaa tgtgctgccg ttttccatcg ccaacctgtc gacctgggtg 960
tcgcatttgc cagcgcttac ggcgacggca tgacgcgtgt gctggaatcg gggttctacg 1020
agcaaatgac ccgtgactcg acgattatcg tcgccaacct gtccgacccg gcgcgcgcca 1080
acacctgggt ccaggacctc aaccgcaatg ccgagccgca cacaggcaat accttcatca 1140
tcggcagcga cggcaatgac ctgatccagg gcggcaaggg cgcggacttc atcgaaggcg 1200
gcaagggcaa tgacacgatc cgcgacaaca gcgggcacaa cacctttttg ttcagcgggc 1260
attttggcca ggatcggatt atcggctacc agccgaccga caggctggtg ttccagggcg 1320
ccgacggcag caccgacctg cgcgaccacg cgaaggccgt gggggccgat acggtgctga 1380
gttttggcgc cgactcggtg accctggtcg gggtcgggct gggcggcctg tggagcgagg 1440
gtgtgctgat cagttgagg 1459
<210> 47
<211> 1489
<212> DNA
<213> Artificial Sequence
<220>
<223> Paracin I - lipase TliA
<400> 47
aaaggaagag gcaaacaggg aggcaaggtg cgtgcgaagg caaagacacg ttcatccatg 60
ggtgtatttg actacaagaa cctcggcacc gaagccagca aaaccttgtt cgccgatgcc 120
accgcaatca cgttgtatac ctatcacaac ctggataacg gcttcgcagt cggctaccag 180
caacatggct tggggctcgg cctgccggcc acactggtcg gggcgttgct cggcagcaca 240
gactcccagg gagtgatccc cggcattccc tggaatcctg actcggaaaa ggccgccctg 300
gacgcggtgc acgcagccgg ttggacgcca atcagcgcca gcgcactggg ctacggcggc 360
aaggtggatg cgcggggcac tttttttggc gagaaggccg gctacaccac ggcccaggcc 420
gaagtgctgg gcaagtacga tgacgccggc aaactgctcg agatcggcat cggttttcgt 480
ggcacctcgg gccctcggga aagcctgatt accgactcca tcggcgatct ggtcagcgac 540
ctgctcgccg cgctgggccc caaggactat gcgaaaaact atgccggcga agcgtttggc 600
ggcttgctca agacggtggc cgactatgcc ggcgcccatg gcctgagtgg caaggatgtg 660
ctggtcagcg gccacagcct gggcggcctg gcggtcaaca gcatggccga cctgagcacc 720
agcaaatggg cgggtttcta caaggacgcc aactacctgg cctacgcctc gcccacccag 780
agcgccggcg ataaggtcct gaatatcggc tacgaaaacg acccggtatt ccgtgcgctg 840
gacggctcca ccttcaacct gtcgtccctc ggcgtgcatg acaaggccca cgagtcgacc 900
accgacaaca tcgtcagctt caacgaccac tacgcctcga cgttgtggaa tgtgctgccg 960
ttttccatcg ccaacctgtc gacctgggtg tcgcatttgc cagcgcttac ggcgacggca 1020
tgacgcgtgt gctggaatcg gggttctacg agcaaatgac ccgtgactcg acgattatcg 1080
tcgccaacct gtccgacccg gcgcgcgcca acacctgggt ccaggacctc aaccgcaatg 1140
ccgagccgca cacaggcaat accttcatca tcggcagcga cggcaatgac ctgatccagg 1200
gcggcaaggg cgcggacttc atcgaaggcg gcaagggcaa tgacacgatc cgcgacaaca 1260
gcgggcacaa cacctttttg ttcagcgggc attttggcca ggatcggatt atcggctacc 1320
agccgaccga caggctggtg ttccagggcg ccgacggcag caccgacctg cgcgaccacg 1380
cgaaggccgt gggggccgat acggtgctga gttttggcgc cgactcggtg accctggtcg 1440
gggtcgggct gggcggcctg tggagcgagg gtgtgctgat cagttgagg 1489
<210> 48
<211> 1489
<212> DNA
<213> Artificial Sequence
<220>
<223> NKC - lipase TliA
<400> 48
gcgccgaaag cgatgaaact gttgaagaaa ttgctgaaat tacagaaaaa aggcattatg 60
ggtgtatttg actacaagaa cctcggcacc gaagccagca aaaccttgtt cgccgatgcc 120
accgcaatca cgttgtatac ctatcacaac ctggataacg gcttcgcagt cggctaccag 180
caacatggct tggggctcgg cctgccggcc acactggtcg gggcgttgct cggcagcaca 240
gactcccagg gagtgatccc cggcattccc tggaatcctg actcggaaaa ggccgccctg 300
gacgcggtgc acgcagccgg ttggacgcca atcagcgcca gcgcactggg ctacggcggc 360
aaggtggatg cgcggggcac tttttttggc gagaaggccg gctacaccac ggcccaggcc 420
gaagtgctgg gcaagtacga tgacgccggc aaactgctcg agatcggcat cggttttcgt 480
ggcacctcgg gccctcggga aagcctgatt accgactcca tcggcgatct ggtcagcgac 540
ctgctcgccg cgctgggccc caaggactat gcgaaaaact atgccggcga agcgtttggc 600
ggcttgctca agacggtggc cgactatgcc ggcgcccatg gcctgagtgg caaggatgtg 660
ctggtcagcg gccacagcct gggcggcctg gcggtcaaca gcatggccga cctgagcacc 720
agcaaatggg cgggtttcta caaggacgcc aactacctgg cctacgcctc gcccacccag 780
agcgccggcg ataaggtcct gaatatcggc tacgaaaacg acccggtatt ccgtgcgctg 840
gacggctcca ccttcaacct gtcgtccctc ggcgtgcatg acaaggccca cgagtcgacc 900
accgacaaca tcgtcagctt caacgaccac tacgcctcga cgttgtggaa tgtgctgccg 960
ttttccatcg ccaacctgtc gacctgggtg tcgcatttgc cagcgcttac ggcgacggca 1020
tgacgcgtgt gctggaatcg gggttctacg agcaaatgac ccgtgactcg acgattatcg 1080
tcgccaacct gtccgacccg gcgcgcgcca acacctgggt ccaggacctc aaccgcaatg 1140
ccgagccgca cacaggcaat accttcatca tcggcagcga cggcaatgac ctgatccagg 1200
gcggcaaggg cgcggacttc atcgaaggcg gcaagggcaa tgacacgatc cgcgacaaca 1260
gcgggcacaa cacctttttg ttcagcgggc attttggcca ggatcggatt atcggctacc 1320
agccgaccga caggctggtg ttccagggcg ccgacggcag caccgacctg cgcgaccacg 1380
cgaaggccgt gggggccgat acggtgctga gttttggcgc cgactcggtg accctggtcg 1440
gggtcgggct gggcggcctg tggagcgagg gtgtgctgat cagttgagg 1489
<210> 49
<211> 1489
<212> DNA
<213> Artificial Sequence
<220>
<223> NRC - lipase TliA
<400> 49
gcgccgaaag cgatgcgtct gttgcgtcgc ttgctgcgtt tacagaaaaa aggcattatg 60
ggtgtatttg actacaagaa cctcggcacc gaagccagca aaaccttgtt cgccgatgcc 120
accgcaatca cgttgtatac ctatcacaac ctggataacg gcttcgcagt cggctaccag 180
caacatggct tggggctcgg cctgccggcc acactggtcg gggcgttgct cggcagcaca 240
gactcccagg gagtgatccc cggcattccc tggaatcctg actcggaaaa ggccgccctg 300
gacgcggtgc acgcagccgg ttggacgcca atcagcgcca gcgcactggg ctacggcggc 360
aaggtggatg cgcggggcac tttttttggc gagaaggccg gctacaccac ggcccaggcc 420
gaagtgctgg gcaagtacga tgacgccggc aaactgctcg agatcggcat cggttttcgt 480
ggcacctcgg gccctcggga aagcctgatt accgactcca tcggcgatct ggtcagcgac 540
ctgctcgccg cgctgggccc caaggactat gcgaaaaact atgccggcga agcgtttggc 600
ggcttgctca agacggtggc cgactatgcc ggcgcccatg gcctgagtgg caaggatgtg 660
ctggtcagcg gccacagcct gggcggcctg gcggtcaaca gcatggccga cctgagcacc 720
agcaaatggg cgggtttcta caaggacgcc aactacctgg cctacgcctc gcccacccag 780
agcgccggcg ataaggtcct gaatatcggc tacgaaaacg acccggtatt ccgtgcgctg 840
gacggctcca ccttcaacct gtcgtccctc ggcgtgcatg acaaggccca cgagtcgacc 900
accgacaaca tcgtcagctt caacgaccac tacgcctcga cgttgtggaa tgtgctgccg 960
ttttccatcg ccaacctgtc gacctgggtg tcgcatttgc cagcgcttac ggcgacggca 1020
tgacgcgtgt gctggaatcg gggttctacg agcaaatgac ccgtgactcg acgattatcg 1080
tcgccaacct gtccgacccg gcgcgcgcca acacctgggt ccaggacctc aaccgcaatg 1140
ccgagccgca cacaggcaat accttcatca tcggcagcga cggcaatgac ctgatccagg 1200
gcggcaaggg cgcggacttc atcgaaggcg gcaagggcaa tgacacgatc cgcgacaaca 1260
gcgggcacaa cacctttttg ttcagcgggc attttggcca ggatcggatt atcggctacc 1320
agccgaccga caggctggtg ttccagggcg ccgacggcag caccgacctg cgcgaccacg 1380
cgaaggccgt gggggccgat acggtgctga gttttggcgc cgactcggtg accctggtcg 1440
gggtcgggct gggcggcctg tggagcgagg gtgtgctgat cagttgagg 1489
<210> 50
<211> 1085
<212> DNA
<213> Artificial Sequence
<220>
<223> Buforin IIb - lipase M37
<400> 50
cgtgctggtc tgcagttccc ggttggtcgt ctgctgcgtc gtctgctgcg tcgtctgctg 60
cgtatgtctt atacaaaaga acaactcatg ttggcattca gctatatgag ctactatggc 120
atcactcaca caggttcagc aaaaaaaaat gctgagctca tccttaaaaa aatgaaagaa 180
gccttgaaaa catggaagcc ttttcaggaa gatgactggg aagtcgtctg gggtcctgcg 240
gtttatacca tgcctttcac aatcttcaac gatgccatga tgtatgtcat acagaagaaa 300
ggtgctgaag gggaatacgt gatagccatt cgcggcacca atccagtatc aatttcagac 360
tggctgttta atgatttcat ggtcagcgca atgaagaagt ggccttacgc atccgttgaa 420
ggccgcatac tcaaaatatc cgaaagtacc agctacggac tgaaaacctt acagaaattg 480
aagccaaaat cccatatccc cggcgaaaat aaaacgattc tgcagttcct gaatgaaaag 540
ataggcccag agggtaaagc aaaaatctgt gtaacaggcc acagtaaagg cggcgccttg 600
tcttccactc tggcactgtg gttgaaggac atccaaggag taaaactctc gcaaaacatc 660
gatatctcaa cgattccgtt tgccggacca acagccggta atgctgactt tgccgattac 720
tttgatgatt gtcttggtga tcaatgcacc cgcattgcca actcgttaga tattgtgcct 780
tatgcctgga ataaaattca ttaaaaaaac ttaaatctat atatatttct gaacaagcat 840
cagttaaacc gcttctatat caacgcgctt taatccgtgc aatgatcgca gaaactaaag 900
gtaaaaaata caagcaaatt aaggcggaaa caccaccatt agaaggcaac attaatccta 960
ttcttattga ataccttgtg caggcagcat atcagcatgt cgtcggttac ccagaattaa 1020
tgggtatgat ggatgatatt cctttaacag acatattcga agatgcgatc gcgggtttgt 1080
tataa 1085
<210> 51
<211> 1049
<212> DNA
<213> Artificial Sequence
<220>
<223> B0 - lipase M37
<400> 51
cgtgctggtc tgcagttccc ggttggtatg tcttatacaa aagaacaact catgttggca 60
ttcagctata tgagctacta tggcatcact cacacaggtt cagcaaaaaa aaatgctgag 120
ctcatcctta aaaaaatgaa agaagccttg aaaacatgga agccttttca ggaagatgac 180
tgggaagtcg tctggggtcc tgcggtttat accatgcctt tcacaatctt caacgatgcc 240
atgatgtatg tcatacagaa gaaaggtgct gaaggggaat acgtgatagc cattcgcggc 300
accaatccag tatcaatttc agactggctg tttaatgatt tcatggtcag cgcaatgaag 360
aagtggcctt acgcatccgt tgaaggccgc atactcaaaa tatccgaaag taccagctac 420
ggactgaaaa ccttacagaa attgaagcca aaatcccata tccccggcga aaataaaacg 480
attctgcagt tcctgaatga aaagataggc ccagagggta aagcaaaaat ctgtgtaaca 540
ggccacagta aaggcggcgc cttgtcttcc actctggcac tgtggttgaa ggacatccaa 600
ggagtaaaac tctcgcaaaa catcgatatc tcaacgattc cgtttgccgg accaacagcc 660
ggtaatgctg actttgccga ttactttgat gattgtcttg gtgatcaatg cacccgcatt 720
gccaactcgt tagatattgt gccttatgcc tggaataaaa ttcattaaaa aaacttaaat 780
ctatatatat ttctgaacaa gcatcagtta aaccgcttct atatcaacgc gctttaatcc 840
gtgcaatgat cgcagaaact aaaggtaaaa aatacaagca aattaaggcg gaaacaccac 900
cattagaagg caacattaat cctattctta ttgaatacct tgtgcaggca gcatatcagc 960
atgtcgtcgg ttacccagaa ttaatgggta tgatggatga tattccttta acagacatat 1020
tcgaagatgc gatcgcgggt ttgttataa 1049
<210> 52
<211> 1079
<212> DNA
<213> Artificial Sequence
<220>
<223> Paracin I - lipase M37
<400> 52
aaaggaagag gcaaacaggg aggcaaggtg cgtgcgaagg caaagacacg ttcatccatg 60
tcttatacaa aagaacaact catgttggca ttcagctata tgagctacta tggcatcact 120
cacacaggtt cagcaaaaaa aaatgctgag ctcatcctta aaaaaatgaa agaagccttg 180
aaaacatgga agccttttca ggaagatgac tgggaagtcg tctggggtcc tgcggtttat 240
accatgcctt tcacaatctt caacgatgcc atgatgtatg tcatacagaa gaaaggtgct 300
gaaggggaat acgtgatagc cattcgcggc accaatccag tatcaatttc agactggctg 360
tttaatgatt tcatggtcag cgcaatgaag aagtggcctt acgcatccgt tgaaggccgc 420
atactcaaaa tatccgaaag taccagctac ggactgaaaa ccttacagaa attgaagcca 480
aaatcccata tccccggcga aaataaaacg attctgcagt tcctgaatga aaagataggc 540
ccagagggta aagcaaaaat ctgtgtaaca ggccacagta aaggcggcgc cttgtcttcc 600
actctggcac tgtggttgaa ggacatccaa ggagtaaaac tctcgcaaaa catcgatatc 660
tcaacgattc cgtttgccgg accaacagcc ggtaatgctg actttgccga ttactttgat 720
gattgtcttg gtgatcaatg cacccgcatt gccaactcgt tagatattgt gccttatgcc 780
tggaataaaa ttcattaaaa aaacttaaat ctatatatat ttctgaacaa gcatcagtta 840
aaccgcttct atatcaacgc gctttaatcc gtgcaatgat cgcagaaact aaaggtaaaa 900
aatacaagca aattaaggcg gaaacaccac cattagaagg caacattaat cctattctta 960
ttgaatacct tgtgcaggca gcatatcagc atgtcgtcgg ttacccagaa ttaatgggta 1020
tgatggatga tattccttta acagacatat tcgaagatgc gatcgcgggt ttgttataa 1079
<210> 53
<211> 1079
<212> DNA
<213> Artificial Sequence
<220>
<223> NKC - lipase M37
<400> 53
gcgccgaaag cgatgaaact gttgaagaaa ttgctgaaat tacagaaaaa aggcattatg 60
tcttatacaa aagaacaact catgttggca ttcagctata tgagctacta tggcatcact 120
cacacaggtt cagcaaaaaa aaatgctgag ctcatcctta aaaaaatgaa agaagccttg 180
aaaacatgga agccttttca ggaagatgac tgggaagtcg tctggggtcc tgcggtttat 240
accatgcctt tcacaatctt caacgatgcc atgatgtatg tcatacagaa gaaaggtgct 300
gaaggggaat acgtgatagc cattcgcggc accaatccag tatcaatttc agactggctg 360
tttaatgatt tcatggtcag cgcaatgaag aagtggcctt acgcatccgt tgaaggccgc 420
atactcaaaa tatccgaaag taccagctac ggactgaaaa ccttacagaa attgaagcca 480
aaatcccata tccccggcga aaataaaacg attctgcagt tcctgaatga aaagataggc 540
ccagagggta aagcaaaaat ctgtgtaaca ggccacagta aaggcggcgc cttgtcttcc 600
actctggcac tgtggttgaa ggacatccaa ggagtaaaac tctcgcaaaa catcgatatc 660
tcaacgattc cgtttgccgg accaacagcc ggtaatgctg actttgccga ttactttgat 720
gattgtcttg gtgatcaatg cacccgcatt gccaactcgt tagatattgt gccttatgcc 780
tggaataaaa ttcattaaaa aaacttaaat ctatatatat ttctgaacaa gcatcagtta 840
aaccgcttct atatcaacgc gctttaatcc gtgcaatgat cgcagaaact aaaggtaaaa 900
aatacaagca aattaaggcg gaaacaccac cattagaagg caacattaat cctattctta 960
ttgaatacct tgtgcaggca gcatatcagc atgtcgtcgg ttacccagaa ttaatgggta 1020
tgatggatga tattccttta acagacatat tcgaagatgc gatcgcgggt ttgttataa 1079
<210> 54
<211> 1079
<212> DNA
<213> Artificial Sequence
<220>
<223> NRC - lipase M37
<400> 54
gcgccgaaag cgatgcgtct gttgcgtcgc ttgctgcgtt tacagaaaaa aggcattatg 60
tcttatacaa aagaacaact catgttggca ttcagctata tgagctacta tggcatcact 120
cacacaggtt cagcaaaaaa aaatgctgag ctcatcctta aaaaaatgaa agaagccttg 180
aaaacatgga agccttttca ggaagatgac tgggaagtcg tctggggtcc tgcggtttat 240
accatgcctt tcacaatctt caacgatgcc atgatgtatg tcatacagaa gaaaggtgct 300
gaaggggaat acgtgatagc cattcgcggc accaatccag tatcaatttc agactggctg 360
tttaatgatt tcatggtcag cgcaatgaag aagtggcctt acgcatccgt tgaaggccgc 420
atactcaaaa tatccgaaag taccagctac ggactgaaaa ccttacagaa attgaagcca 480
aaatcccata tccccggcga aaataaaacg attctgcagt tcctgaatga aaagataggc 540
ccagagggta aagcaaaaat ctgtgtaaca ggccacagta aaggcggcgc cttgtcttcc 600
actctggcac tgtggttgaa ggacatccaa ggagtaaaac tctcgcaaaa catcgatatc 660
tcaacgattc cgtttgccgg accaacagcc ggtaatgctg actttgccga ttactttgat 720
gattgtcttg gtgatcaatg cacccgcatt gccaactcgt tagatattgt gccttatgcc 780
tggaataaaa ttcattaaaa aaacttaaat ctatatatat ttctgaacaa gcatcagtta 840
aaccgcttct atatcaacgc gctttaatcc gtgcaatgat cgcagaaact aaaggtaaaa 900
aatacaagca aattaaggcg gaaacaccac cattagaagg caacattaat cctattctta 960
ttgaatacct tgtgcaggca gcatatcagc atgtcgtcgg ttacccagaa ttaatgggta 1020
tgatggatga tattccttta acagacatat tcgaagatgc gatcgcgggt ttgttataa 1079
<210> 55
<211> 1495
<212> DNA
<213> Artificial Sequence
<220>
<223> lipase TliA - Buforin IIb
<400> 55
atgggtgtat ttgactacaa gaacctcggc accgaagcca gcaaaacctt gttcgccgat 60
gccaccgcaa tcacgttgta tacctatcac aacctggata acggcttcgc agtcggctac 120
cagcaacatg gcttggggct cggcctgccg gccacactgg tcggggcgtt gctcggcagc 180
acagactccc agggagtgat ccccggcatt ccctggaatc ctgactcgga aaaggccgcc 240
ctggacgcgg tgcacgcagc cggttggacg ccaatcagcg ccagcgcact gggctacggc 300
ggcaaggtgg atgcgcgggg cacttttttt ggcgagaagg ccggctacac cacggcccag 360
gccgaagtgc tgggcaagta cgatgacgcc ggcaaactgc tcgagatcgg catcggtttt 420
cgtggcacct cgggccctcg ggaaagcctg attaccgact ccatcggcga tctggtcagc 480
gacctgctcg ccgcgctggg ccccaaggac tatgcgaaaa actatgccgg cgaagcgttt 540
ggcggcttgc tcaagacggt ggccgactat gccggcgccc atggcctgag tggcaaggat 600
gtgctggtca gcggccacag cctgggcggc ctggcggtca acagcatggc cgacctgagc 660
accagcaaat gggcgggttt ctacaaggac gccaactacc tggcctacgc ctcgcccacc 720
cagagcgccg gcgataaggt cctgaatatc ggctacgaaa acgacccggt attccgtgcg 780
ctggacggct ccaccttcaa cctgtcgtcc ctcggcgtgc atgacaaggc ccacgagtcg 840
accaccgaca acatcgtcag cttcaacgac cactacgcct cgacgttgtg gaatgtgctg 900
ccgttttcca tcgccaacct gtcgacctgg gtgtcgcatt tgccagcgct tacggcgacg 960
gcatgacgcg tgtgctggaa tcggggttct acgagcaaat gacccgtgac tcgacgatta 1020
tcgtcgccaa cctgtccgac ccggcgcgcg ccaacacctg ggtccaggac ctcaaccgca 1080
atgccgagcc gcacacaggc aataccttca tcatcggcag cgacggcaat gacctgatcc 1140
agggcggcaa gggcgcggac ttcatcgaag gcggcaaggg caatgacacg atccgcgaca 1200
acagcgggca caacaccttt ttgttcagcg ggcattttgg ccaggatcgg attatcggct 1260
accagccgac cgacaggctg gtgttccagg gcgccgacgg cagcaccgac ctgcgcgacc 1320
acgcgaaggc cgtgggggcc gatacggtgc tgagttttgg cgccgactcg gtgaccctgg 1380
tcggggtcgg gctgggcggc ctgtggagcg agggtgtgct gatcagttga ggcgtgctgg 1440
tctgcagttc ccggttggtc gtctgctgcg tcgtctgctg cgtcgtctgc tgcgt 1495
<210> 56
<211> 1459
<212> DNA
<213> Artificial Sequence
<220>
<223> lipase TliA - B0
<400> 56
atgggtgtat ttgactacaa gaacctcggc accgaagcca gcaaaacctt gttcgccgat 60
gccaccgcaa tcacgttgta tacctatcac aacctggata acggcttcgc agtcggctac 120
cagcaacatg gcttggggct cggcctgccg gccacactgg tcggggcgtt gctcggcagc 180
acagactccc agggagtgat ccccggcatt ccctggaatc ctgactcgga aaaggccgcc 240
ctggacgcgg tgcacgcagc cggttggacg ccaatcagcg ccagcgcact gggctacggc 300
ggcaaggtgg atgcgcgggg cacttttttt ggcgagaagg ccggctacac cacggcccag 360
gccgaagtgc tgggcaagta cgatgacgcc ggcaaactgc tcgagatcgg catcggtttt 420
cgtggcacct cgggccctcg ggaaagcctg attaccgact ccatcggcga tctggtcagc 480
gacctgctcg ccgcgctggg ccccaaggac tatgcgaaaa actatgccgg cgaagcgttt 540
ggcggcttgc tcaagacggt ggccgactat gccggcgccc atggcctgag tggcaaggat 600
gtgctggtca gcggccacag cctgggcggc ctggcggtca acagcatggc cgacctgagc 660
accagcaaat gggcgggttt ctacaaggac gccaactacc tggcctacgc ctcgcccacc 720
cagagcgccg gcgataaggt cctgaatatc ggctacgaaa acgacccggt attccgtgcg 780
ctggacggct ccaccttcaa cctgtcgtcc ctcggcgtgc atgacaaggc ccacgagtcg 840
accaccgaca acatcgtcag cttcaacgac cactacgcct cgacgttgtg gaatgtgctg 900
ccgttttcca tcgccaacct gtcgacctgg gtgtcgcatt tgccagcgct tacggcgacg 960
gcatgacgcg tgtgctggaa tcggggttct acgagcaaat gacccgtgac tcgacgatta 1020
tcgtcgccaa cctgtccgac ccggcgcgcg ccaacacctg ggtccaggac ctcaaccgca 1080
atgccgagcc gcacacaggc aataccttca tcatcggcag cgacggcaat gacctgatcc 1140
agggcggcaa gggcgcggac ttcatcgaag gcggcaaggg caatgacacg atccgcgaca 1200
acagcgggca caacaccttt ttgttcagcg ggcattttgg ccaggatcgg attatcggct 1260
accagccgac cgacaggctg gtgttccagg gcgccgacgg cagcaccgac ctgcgcgacc 1320
acgcgaaggc cgtgggggcc gatacggtgc tgagttttgg cgccgactcg gtgaccctgg 1380
tcggggtcgg gctgggcggc ctgtggagcg agggtgtgct gatcagttga ggcgtgctgg 1440
tctgcagttc ccggttggt 1459
<210> 57
<211> 1489
<212> DNA
<213> Artificial Sequence
<220>
<223> lipase TliA - Paracin I
<400> 57
atgggtgtat ttgactacaa gaacctcggc accgaagcca gcaaaacctt gttcgccgat 60
gccaccgcaa tcacgttgta tacctatcac aacctggata acggcttcgc agtcggctac 120
cagcaacatg gcttggggct cggcctgccg gccacactgg tcggggcgtt gctcggcagc 180
acagactccc agggagtgat ccccggcatt ccctggaatc ctgactcgga aaaggccgcc 240
ctggacgcgg tgcacgcagc cggttggacg ccaatcagcg ccagcgcact gggctacggc 300
ggcaaggtgg atgcgcgggg cacttttttt ggcgagaagg ccggctacac cacggcccag 360
gccgaagtgc tgggcaagta cgatgacgcc ggcaaactgc tcgagatcgg catcggtttt 420
cgtggcacct cgggccctcg ggaaagcctg attaccgact ccatcggcga tctggtcagc 480
gacctgctcg ccgcgctggg ccccaaggac tatgcgaaaa actatgccgg cgaagcgttt 540
ggcggcttgc tcaagacggt ggccgactat gccggcgccc atggcctgag tggcaaggat 600
gtgctggtca gcggccacag cctgggcggc ctggcggtca acagcatggc cgacctgagc 660
accagcaaat gggcgggttt ctacaaggac gccaactacc tggcctacgc ctcgcccacc 720
cagagcgccg gcgataaggt cctgaatatc ggctacgaaa acgacccggt attccgtgcg 780
ctggacggct ccaccttcaa cctgtcgtcc ctcggcgtgc atgacaaggc ccacgagtcg 840
accaccgaca acatcgtcag cttcaacgac cactacgcct cgacgttgtg gaatgtgctg 900
ccgttttcca tcgccaacct gtcgacctgg gtgtcgcatt tgccagcgct tacggcgacg 960
gcatgacgcg tgtgctggaa tcggggttct acgagcaaat gacccgtgac tcgacgatta 1020
tcgtcgccaa cctgtccgac ccggcgcgcg ccaacacctg ggtccaggac ctcaaccgca 1080
atgccgagcc gcacacaggc aataccttca tcatcggcag cgacggcaat gacctgatcc 1140
agggcggcaa gggcgcggac ttcatcgaag gcggcaaggg caatgacacg atccgcgaca 1200
acagcgggca caacaccttt ttgttcagcg ggcattttgg ccaggatcgg attatcggct 1260
accagccgac cgacaggctg gtgttccagg gcgccgacgg cagcaccgac ctgcgcgacc 1320
acgcgaaggc cgtgggggcc gatacggtgc tgagttttgg cgccgactcg gtgaccctgg 1380
tcggggtcgg gctgggcggc ctgtggagcg agggtgtgct gatcagttga ggaaaggaag 1440
aggcaaacag ggaggcaagg tgcgtgcgaa ggcaaagaca cgttcatcc 1489
<210> 58
<211> 1489
<212> DNA
<213> Artificial Sequence
<220>
<223> lipase TliA - NRC
<400> 58
atgggtgtat ttgactacaa gaacctcggc accgaagcca gcaaaacctt gttcgccgat 60
gccaccgcaa tcacgttgta tacctatcac aacctggata acggcttcgc agtcggctac 120
cagcaacatg gcttggggct cggcctgccg gccacactgg tcggggcgtt gctcggcagc 180
acagactccc agggagtgat ccccggcatt ccctggaatc ctgactcgga aaaggccgcc 240
ctggacgcgg tgcacgcagc cggttggacg ccaatcagcg ccagcgcact gggctacggc 300
ggcaaggtgg atgcgcgggg cacttttttt ggcgagaagg ccggctacac cacggcccag 360
gccgaagtgc tgggcaagta cgatgacgcc ggcaaactgc tcgagatcgg catcggtttt 420
cgtggcacct cgggccctcg ggaaagcctg attaccgact ccatcggcga tctggtcagc 480
gacctgctcg ccgcgctggg ccccaaggac tatgcgaaaa actatgccgg cgaagcgttt 540
ggcggcttgc tcaagacggt ggccgactat gccggcgccc atggcctgag tggcaaggat 600
gtgctggtca gcggccacag cctgggcggc ctggcggtca acagcatggc cgacctgagc 660
accagcaaat gggcgggttt ctacaaggac gccaactacc tggcctacgc ctcgcccacc 720
cagagcgccg gcgataaggt cctgaatatc ggctacgaaa acgacccggt attccgtgcg 780
ctggacggct ccaccttcaa cctgtcgtcc ctcggcgtgc atgacaaggc ccacgagtcg 840
accaccgaca acatcgtcag cttcaacgac cactacgcct cgacgttgtg gaatgtgctg 900
ccgttttcca tcgccaacct gtcgacctgg gtgtcgcatt tgccagcgct tacggcgacg 960
gcatgacgcg tgtgctggaa tcggggttct acgagcaaat gacccgtgac tcgacgatta 1020
tcgtcgccaa cctgtccgac ccggcgcgcg ccaacacctg ggtccaggac ctcaaccgca 1080
atgccgagcc gcacacaggc aataccttca tcatcggcag cgacggcaat gacctgatcc 1140
agggcggcaa gggcgcggac ttcatcgaag gcggcaaggg caatgacacg atccgcgaca 1200
acagcgggca caacaccttt ttgttcagcg ggcattttgg ccaggatcgg attatcggct 1260
accagccgac cgacaggctg gtgttccagg gcgccgacgg cagcaccgac ctgcgcgacc 1320
acgcgaaggc cgtgggggcc gatacggtgc tgagttttgg cgccgactcg gtgaccctgg 1380
tcggggtcgg gctgggcggc ctgtggagcg agggtgtgct gatcagttga gggcgccgaa 1440
agcgatgaaa ctgttgaaga aattgctgaa attacagaaa aaaggcatt 1489
<210> 59
<211> 1489
<212> DNA
<213> Artificial Sequence
<220>
<223> lipase TliA - NRC
<400> 59
atgggtgtat ttgactacaa gaacctcggc accgaagcca gcaaaacctt gttcgccgat 60
gccaccgcaa tcacgttgta tacctatcac aacctggata acggcttcgc agtcggctac 120
cagcaacatg gcttggggct cggcctgccg gccacactgg tcggggcgtt gctcggcagc 180
acagactccc agggagtgat ccccggcatt ccctggaatc ctgactcgga aaaggccgcc 240
ctggacgcgg tgcacgcagc cggttggacg ccaatcagcg ccagcgcact gggctacggc 300
ggcaaggtgg atgcgcgggg cacttttttt ggcgagaagg ccggctacac cacggcccag 360
gccgaagtgc tgggcaagta cgatgacgcc ggcaaactgc tcgagatcgg catcggtttt 420
cgtggcacct cgggccctcg ggaaagcctg attaccgact ccatcggcga tctggtcagc 480
gacctgctcg ccgcgctggg ccccaaggac tatgcgaaaa actatgccgg cgaagcgttt 540
ggcggcttgc tcaagacggt ggccgactat gccggcgccc atggcctgag tggcaaggat 600
gtgctggtca gcggccacag cctgggcggc ctggcggtca acagcatggc cgacctgagc 660
accagcaaat gggcgggttt ctacaaggac gccaactacc tggcctacgc ctcgcccacc 720
cagagcgccg gcgataaggt cctgaatatc ggctacgaaa acgacccggt attccgtgcg 780
ctggacggct ccaccttcaa cctgtcgtcc ctcggcgtgc atgacaaggc ccacgagtcg 840
accaccgaca acatcgtcag cttcaacgac cactacgcct cgacgttgtg gaatgtgctg 900
ccgttttcca tcgccaacct gtcgacctgg gtgtcgcatt tgccagcgct tacggcgacg 960
gcatgacgcg tgtgctggaa tcggggttct acgagcaaat gacccgtgac tcgacgatta 1020
tcgtcgccaa cctgtccgac ccggcgcgcg ccaacacctg ggtccaggac ctcaaccgca 1080
atgccgagcc gcacacaggc aataccttca tcatcggcag cgacggcaat gacctgatcc 1140
agggcggcaa gggcgcggac ttcatcgaag gcggcaaggg caatgacacg atccgcgaca 1200
acagcgggca caacaccttt ttgttcagcg ggcattttgg ccaggatcgg attatcggct 1260
accagccgac cgacaggctg gtgttccagg gcgccgacgg cagcaccgac ctgcgcgacc 1320
acgcgaaggc cgtgggggcc gatacggtgc tgagttttgg cgccgactcg gtgaccctgg 1380
tcggggtcgg gctgggcggc ctgtggagcg agggtgtgct gatcagttga gggcgccgaa 1440
agcgatgcgt ctgttgcgtc gcttgctgcg tttacagaaa aaaggcatt 1489
<210> 60
<211> 1085
<212> DNA
<213> Artificial Sequence
<220>
<223> lipase M37 - Buforin IIb
<400> 60
atgtcttata caaaagaaca actcatgttg gcattcagct atatgagcta ctatggcatc 60
actcacacag gttcagcaaa aaaaaatgct gagctcatcc ttaaaaaaat gaaagaagcc 120
ttgaaaacat ggaagccttt tcaggaagat gactgggaag tcgtctgggg tcctgcggtt 180
tataccatgc ctttcacaat cttcaacgat gccatgatgt atgtcataca gaagaaaggt 240
gctgaagggg aatacgtgat agccattcgc ggcaccaatc cagtatcaat ttcagactgg 300
ctgtttaatg atttcatggt cagcgcaatg aagaagtggc cttacgcatc cgttgaaggc 360
cgcatactca aaatatccga aagtaccagc tacggactga aaaccttaca gaaattgaag 420
ccaaaatccc atatccccgg cgaaaataaa acgattctgc agttcctgaa tgaaaagata 480
ggcccagagg gtaaagcaaa aatctgtgta acaggccaca gtaaaggcgg cgccttgtct 540
tccactctgg cactgtggtt gaaggacatc caaggagtaa aactctcgca aaacatcgat 600
atctcaacga ttccgtttgc cggaccaaca gccggtaatg ctgactttgc cgattacttt 660
gatgattgtc ttggtgatca atgcacccgc attgccaact cgttagatat tgtgccttat 720
gcctggaata aaattcatta aaaaaactta aatctatata tatttctgaa caagcatcag 780
ttaaaccgct tctatatcaa cgcgctttaa tccgtgcaat gatcgcagaa actaaaggta 840
aaaaatacaa gcaaattaag gcggaaacac caccattaga aggcaacatt aatcctattc 900
ttattgaata ccttgtgcag gcagcatatc agcatgtcgt cggttaccca gaattaatgg 960
gtatgatgga tgatattcct ttaacagaca tattcgaaga tgcgatcgcg ggtttgttat 1020
aacgtgctgg tctgcagttc ccggttggtc gtctgctgcg tcgtctgctg cgtcgtctgc 1080
tgcgt 1085
<210> 61
<211> 1049
<212> DNA
<213> Artificial Sequence
<220>
<223> lipase M37 - B0
<400> 61
atgtcttata caaaagaaca actcatgttg gcattcagct atatgagcta ctatggcatc 60
actcacacag gttcagcaaa aaaaaatgct gagctcatcc ttaaaaaaat gaaagaagcc 120
ttgaaaacat ggaagccttt tcaggaagat gactgggaag tcgtctgggg tcctgcggtt 180
tataccatgc ctttcacaat cttcaacgat gccatgatgt atgtcataca gaagaaaggt 240
gctgaagggg aatacgtgat agccattcgc ggcaccaatc cagtatcaat ttcagactgg 300
ctgtttaatg atttcatggt cagcgcaatg aagaagtggc cttacgcatc cgttgaaggc 360
cgcatactca aaatatccga aagtaccagc tacggactga aaaccttaca gaaattgaag 420
ccaaaatccc atatccccgg cgaaaataaa acgattctgc agttcctgaa tgaaaagata 480
ggcccagagg gtaaagcaaa aatctgtgta acaggccaca gtaaaggcgg cgccttgtct 540
tccactctgg cactgtggtt gaaggacatc caaggagtaa aactctcgca aaacatcgat 600
atctcaacga ttccgtttgc cggaccaaca gccggtaatg ctgactttgc cgattacttt 660
gatgattgtc ttggtgatca atgcacccgc attgccaact cgttagatat tgtgccttat 720
gcctggaata aaattcatta aaaaaactta aatctatata tatttctgaa caagcatcag 780
ttaaaccgct tctatatcaa cgcgctttaa tccgtgcaat gatcgcagaa actaaaggta 840
aaaaatacaa gcaaattaag gcggaaacac caccattaga aggcaacatt aatcctattc 900
ttattgaata ccttgtgcag gcagcatatc agcatgtcgt cggttaccca gaattaatgg 960
gtatgatgga tgatattcct ttaacagaca tattcgaaga tgcgatcgcg ggtttgttat 1020
aacgtgctgg tctgcagttc ccggttggt 1049
<210> 62
<211> 1079
<212> DNA
<213> Artificial Sequence
<220>
<223> lipase M37 - Paracin I
<400> 62
atgtcttata caaaagaaca actcatgttg gcattcagct atatgagcta ctatggcatc 60
actcacacag gttcagcaaa aaaaaatgct gagctcatcc ttaaaaaaat gaaagaagcc 120
ttgaaaacat ggaagccttt tcaggaagat gactgggaag tcgtctgggg tcctgcggtt 180
tataccatgc ctttcacaat cttcaacgat gccatgatgt atgtcataca gaagaaaggt 240
gctgaagggg aatacgtgat agccattcgc ggcaccaatc cagtatcaat ttcagactgg 300
ctgtttaatg atttcatggt cagcgcaatg aagaagtggc cttacgcatc cgttgaaggc 360
cgcatactca aaatatccga aagtaccagc tacggactga aaaccttaca gaaattgaag 420
ccaaaatccc atatccccgg cgaaaataaa acgattctgc agttcctgaa tgaaaagata 480
ggcccagagg gtaaagcaaa aatctgtgta acaggccaca gtaaaggcgg cgccttgtct 540
tccactctgg cactgtggtt gaaggacatc caaggagtaa aactctcgca aaacatcgat 600
atctcaacga ttccgtttgc cggaccaaca gccggtaatg ctgactttgc cgattacttt 660
gatgattgtc ttggtgatca atgcacccgc attgccaact cgttagatat tgtgccttat 720
gcctggaata aaattcatta aaaaaactta aatctatata tatttctgaa caagcatcag 780
ttaaaccgct tctatatcaa cgcgctttaa tccgtgcaat gatcgcagaa actaaaggta 840
aaaaatacaa gcaaattaag gcggaaacac caccattaga aggcaacatt aatcctattc 900
ttattgaata ccttgtgcag gcagcatatc agcatgtcgt cggttaccca gaattaatgg 960
gtatgatgga tgatattcct ttaacagaca tattcgaaga tgcgatcgcg ggtttgttat 1020
aaaaaggaag aggcaaacag ggaggcaagg tgcgtgcgaa ggcaaagaca cgttcatcc 1079
<210> 63
<211> 1079
<212> DNA
<213> Artificial Sequence
<220>
<223> lipase M37 - NKC
<400> 63
atgtcttata caaaagaaca actcatgttg gcattcagct atatgagcta ctatggcatc 60
actcacacag gttcagcaaa aaaaaatgct gagctcatcc ttaaaaaaat gaaagaagcc 120
ttgaaaacat ggaagccttt tcaggaagat gactgggaag tcgtctgggg tcctgcggtt 180
tataccatgc ctttcacaat cttcaacgat gccatgatgt atgtcataca gaagaaaggt 240
gctgaagggg aatacgtgat agccattcgc ggcaccaatc cagtatcaat ttcagactgg 300
ctgtttaatg atttcatggt cagcgcaatg aagaagtggc cttacgcatc cgttgaaggc 360
cgcatactca aaatatccga aagtaccagc tacggactga aaaccttaca gaaattgaag 420
ccaaaatccc atatccccgg cgaaaataaa acgattctgc agttcctgaa tgaaaagata 480
ggcccagagg gtaaagcaaa aatctgtgta acaggccaca gtaaaggcgg cgccttgtct 540
tccactctgg cactgtggtt gaaggacatc caaggagtaa aactctcgca aaacatcgat 600
atctcaacga ttccgtttgc cggaccaaca gccggtaatg ctgactttgc cgattacttt 660
gatgattgtc ttggtgatca atgcacccgc attgccaact cgttagatat tgtgccttat 720
gcctggaata aaattcatta aaaaaactta aatctatata tatttctgaa caagcatcag 780
ttaaaccgct tctatatcaa cgcgctttaa tccgtgcaat gatcgcagaa actaaaggta 840
aaaaatacaa gcaaattaag gcggaaacac caccattaga aggcaacatt aatcctattc 900
ttattgaata ccttgtgcag gcagcatatc agcatgtcgt cggttaccca gaattaatgg 960
gtatgatgga tgatattcct ttaacagaca tattcgaaga tgcgatcgcg ggtttgttat 1020
aagcgccgaa agcgatgaaa ctgttgaaga aattgctgaa attacagaaa aaaggcatt 1079
<210> 64
<211> 1079
<212> DNA
<213> Artificial Sequence
<220>
<223> lipase M37 - NRC
<400> 64
atgtcttata caaaagaaca actcatgttg gcattcagct atatgagcta ctatggcatc 60
actcacacag gttcagcaaa aaaaaatgct gagctcatcc ttaaaaaaat gaaagaagcc 120
ttgaaaacat ggaagccttt tcaggaagat gactgggaag tcgtctgggg tcctgcggtt 180
tataccatgc ctttcacaat cttcaacgat gccatgatgt atgtcataca gaagaaaggt 240
gctgaagggg aatacgtgat agccattcgc ggcaccaatc cagtatcaat ttcagactgg 300
ctgtttaatg atttcatggt cagcgcaatg aagaagtggc cttacgcatc cgttgaaggc 360
cgcatactca aaatatccga aagtaccagc tacggactga aaaccttaca gaaattgaag 420
ccaaaatccc atatccccgg cgaaaataaa acgattctgc agttcctgaa tgaaaagata 480
ggcccagagg gtaaagcaaa aatctgtgta acaggccaca gtaaaggcgg cgccttgtct 540
tccactctgg cactgtggtt gaaggacatc caaggagtaa aactctcgca aaacatcgat 600
atctcaacga ttccgtttgc cggaccaaca gccggtaatg ctgactttgc cgattacttt 660
gatgattgtc ttggtgatca atgcacccgc attgccaact cgttagatat tgtgccttat 720
gcctggaata aaattcatta aaaaaactta aatctatata tatttctgaa caagcatcag 780
ttaaaccgct tctatatcaa cgcgctttaa tccgtgcaat gatcgcagaa actaaaggta 840
aaaaatacaa gcaaattaag gcggaaacac caccattaga aggcaacatt aatcctattc 900
ttattgaata ccttgtgcag gcagcatatc agcatgtcgt cggttaccca gaattaatgg 960
gtatgatgga tgatattcct ttaacagaca tattcgaaga tgcgatcgcg ggtttgttat 1020
aagcgccgaa agcgatgcgt ctgttgcgtc gcttgctgcg tttacagaaa aaaggcatt 1079
<210> 65
<211> 1558
<212> DNA
<213> Artificial Sequence
<220>
<223> Buforin IIb - lipase TliA - Buforin IIb
<400> 65
cgtgctggtc tgcagttccc ggttggtcgt ctgctgcgtc gtctgctgcg tcgtctgctg 60
cgtatgggtg tatttgacta caagaacctc ggcaccgaag ccagcaaaac cttgttcgcc 120
gatgccaccg caatcacgtt gtatacctat cacaacctgg ataacggctt cgcagtcggc 180
taccagcaac atggcttggg gctcggcctg ccggccacac tggtcggggc gttgctcggc 240
agcacagact cccagggagt gatccccggc attccctgga atcctgactc ggaaaaggcc 300
gccctggacg cggtgcacgc agccggttgg acgccaatca gcgccagcgc actgggctac 360
ggcggcaagg tggatgcgcg gggcactttt tttggcgaga aggccggcta caccacggcc 420
caggccgaag tgctgggcaa gtacgatgac gccggcaaac tgctcgagat cggcatcggt 480
tttcgtggca cctcgggccc tcgggaaagc ctgattaccg actccatcgg cgatctggtc 540
agcgacctgc tcgccgcgct gggccccaag gactatgcga aaaactatgc cggcgaagcg 600
tttggcggct tgctcaagac ggtggccgac tatgccggcg cccatggcct gagtggcaag 660
gatgtgctgg tcagcggcca cagcctgggc ggcctggcgg tcaacagcat ggccgacctg 720
agcaccagca aatgggcggg tttctacaag gacgccaact acctggccta cgcctcgccc 780
acccagagcg ccggcgataa ggtcctgaat atcggctacg aaaacgaccc ggtattccgt 840
gcgctggacg gctccacctt caacctgtcg tccctcggcg tgcatgacaa ggcccacgag 900
tcgaccaccg acaacatcgt cagcttcaac gaccactacg cctcgacgtt gtggaatgtg 960
ctgccgtttt ccatcgccaa cctgtcgacc tgggtgtcgc atttgccagc gcttacggcg 1020
acggcatgac gcgtgtgctg gaatcggggt tctacgagca aatgacccgt gactcgacga 1080
ttatcgtcgc caacctgtcc gacccggcgc gcgccaacac ctgggtccag gacctcaacc 1140
gcaatgccga gccgcacaca ggcaatacct tcatcatcgg cagcgacggc aatgacctga 1200
tccagggcgg caagggcgcg gacttcatcg aaggcggcaa gggcaatgac acgatccgcg 1260
acaacagcgg gcacaacacc tttttgttca gcgggcattt tggccaggat cggattatcg 1320
gctaccagcc gaccgacagg ctggtgttcc agggcgccga cggcagcacc gacctgcgcg 1380
accacgcgaa ggccgtgggg gccgatacgg tgctgagttt tggcgccgac tcggtgaccc 1440
tggtcggggt cgggctgggc ggcctgtgga gcgagggtgt gctgatcagt tgaggcgtgc 1500
tggtctgcag ttcccggttg gtcgtctgct gcgtcgtctg ctgcgtcgtc tgctgcgt 1558
<210> 66
<211> 1486
<212> DNA
<213> Artificial Sequence
<220>
<223> B0 - lipase TliA - B0
<400> 66
cgtgctggtc tgcagttccc ggttggtatg ggtgtatttg actacaagaa cctcggcacc 60
gaagccagca aaaccttgtt cgccgatgcc accgcaatca cgttgtatac ctatcacaac 120
ctggataacg gcttcgcagt cggctaccag caacatggct tggggctcgg cctgccggcc 180
acactggtcg gggcgttgct cggcagcaca gactcccagg gagtgatccc cggcattccc 240
tggaatcctg actcggaaaa ggccgccctg gacgcggtgc acgcagccgg ttggacgcca 300
atcagcgcca gcgcactggg ctacggcggc aaggtggatg cgcggggcac tttttttggc 360
gagaaggccg gctacaccac ggcccaggcc gaagtgctgg gcaagtacga tgacgccggc 420
aaactgctcg agatcggcat cggttttcgt ggcacctcgg gccctcggga aagcctgatt 480
accgactcca tcggcgatct ggtcagcgac ctgctcgccg cgctgggccc caaggactat 540
gcgaaaaact atgccggcga agcgtttggc ggcttgctca agacggtggc cgactatgcc 600
ggcgcccatg gcctgagtgg caaggatgtg ctggtcagcg gccacagcct gggcggcctg 660
gcggtcaaca gcatggccga cctgagcacc agcaaatggg cgggtttcta caaggacgcc 720
aactacctgg cctacgcctc gcccacccag agcgccggcg ataaggtcct gaatatcggc 780
tacgaaaacg acccggtatt ccgtgcgctg gacggctcca ccttcaacct gtcgtccctc 840
ggcgtgcatg acaaggccca cgagtcgacc accgacaaca tcgtcagctt caacgaccac 900
tacgcctcga cgttgtggaa tgtgctgccg ttttccatcg ccaacctgtc gacctgggtg 960
tcgcatttgc cagcgcttac ggcgacggca tgacgcgtgt gctggaatcg gggttctacg 1020
agcaaatgac ccgtgactcg acgattatcg tcgccaacct gtccgacccg gcgcgcgcca 1080
acacctgggt ccaggacctc aaccgcaatg ccgagccgca cacaggcaat accttcatca 1140
tcggcagcga cggcaatgac ctgatccagg gcggcaaggg cgcggacttc atcgaaggcg 1200
gcaagggcaa tgacacgatc cgcgacaaca gcgggcacaa cacctttttg ttcagcgggc 1260
attttggcca ggatcggatt atcggctacc agccgaccga caggctggtg ttccagggcg 1320
ccgacggcag caccgacctg cgcgaccacg cgaaggccgt gggggccgat acggtgctga 1380
gttttggcgc cgactcggtg accctggtcg gggtcgggct gggcggcctg tggagcgagg 1440
gtgtgctgat cagttgaggc gtgctggtct gcagttcccg gttggt 1486
<210> 67
<211> 1546
<212> DNA
<213> Artificial Sequence
<220>
<223> Paracin I - lipase TliA - Paracin I
<400> 67
aaaggaagag gcaaacaggg aggcaaggtg cgtgcgaagg caaagacacg ttcatccatg 60
ggtgtatttg actacaagaa cctcggcacc gaagccagca aaaccttgtt cgccgatgcc 120
accgcaatca cgttgtatac ctatcacaac ctggataacg gcttcgcagt cggctaccag 180
caacatggct tggggctcgg cctgccggcc acactggtcg gggcgttgct cggcagcaca 240
gactcccagg gagtgatccc cggcattccc tggaatcctg actcggaaaa ggccgccctg 300
gacgcggtgc acgcagccgg ttggacgcca atcagcgcca gcgcactggg ctacggcggc 360
aaggtggatg cgcggggcac tttttttggc gagaaggccg gctacaccac ggcccaggcc 420
gaagtgctgg gcaagtacga tgacgccggc aaactgctcg agatcggcat cggttttcgt 480
ggcacctcgg gccctcggga aagcctgatt accgactcca tcggcgatct ggtcagcgac 540
ctgctcgccg cgctgggccc caaggactat gcgaaaaact atgccggcga agcgtttggc 600
ggcttgctca agacggtggc cgactatgcc ggcgcccatg gcctgagtgg caaggatgtg 660
ctggtcagcg gccacagcct gggcggcctg gcggtcaaca gcatggccga cctgagcacc 720
agcaaatggg cgggtttcta caaggacgcc aactacctgg cctacgcctc gcccacccag 780
agcgccggcg ataaggtcct gaatatcggc tacgaaaacg acccggtatt ccgtgcgctg 840
gacggctcca ccttcaacct gtcgtccctc ggcgtgcatg acaaggccca cgagtcgacc 900
accgacaaca tcgtcagctt caacgaccac tacgcctcga cgttgtggaa tgtgctgccg 960
ttttccatcg ccaacctgtc gacctgggtg tcgcatttgc cagcgcttac ggcgacggca 1020
tgacgcgtgt gctggaatcg gggttctacg agcaaatgac ccgtgactcg acgattatcg 1080
tcgccaacct gtccgacccg gcgcgcgcca acacctgggt ccaggacctc aaccgcaatg 1140
ccgagccgca cacaggcaat accttcatca tcggcagcga cggcaatgac ctgatccagg 1200
gcggcaaggg cgcggacttc atcgaaggcg gcaagggcaa tgacacgatc cgcgacaaca 1260
gcgggcacaa cacctttttg ttcagcgggc attttggcca ggatcggatt atcggctacc 1320
agccgaccga caggctggtg ttccagggcg ccgacggcag caccgacctg cgcgaccacg 1380
cgaaggccgt gggggccgat acggtgctga gttttggcgc cgactcggtg accctggtcg 1440
gggtcgggct gggcggcctg tggagcgagg gtgtgctgat cagttgagga aaggaagagg 1500
caaacaggga ggcaaggtgc gtgcgaaggc aaagacacgt tcatcc 1546
<210> 68
<211> 1546
<212> DNA
<213> Artificial Sequence
<220>
<223> NKC - lipase TliA - NKC
<400> 68
gcgccgaaag cgatgaaact gttgaagaaa ttgctgaaat tacagaaaaa aggcattatg 60
ggtgtatttg actacaagaa cctcggcacc gaagccagca aaaccttgtt cgccgatgcc 120
accgcaatca cgttgtatac ctatcacaac ctggataacg gcttcgcagt cggctaccag 180
caacatggct tggggctcgg cctgccggcc acactggtcg gggcgttgct cggcagcaca 240
gactcccagg gagtgatccc cggcattccc tggaatcctg actcggaaaa ggccgccctg 300
gacgcggtgc acgcagccgg ttggacgcca atcagcgcca gcgcactggg ctacggcggc 360
aaggtggatg cgcggggcac tttttttggc gagaaggccg gctacaccac ggcccaggcc 420
gaagtgctgg gcaagtacga tgacgccggc aaactgctcg agatcggcat cggttttcgt 480
ggcacctcgg gccctcggga aagcctgatt accgactcca tcggcgatct ggtcagcgac 540
ctgctcgccg cgctgggccc caaggactat gcgaaaaact atgccggcga agcgtttggc 600
ggcttgctca agacggtggc cgactatgcc ggcgcccatg gcctgagtgg caaggatgtg 660
ctggtcagcg gccacagcct gggcggcctg gcggtcaaca gcatggccga cctgagcacc 720
agcaaatggg cgggtttcta caaggacgcc aactacctgg cctacgcctc gcccacccag 780
agcgccggcg ataaggtcct gaatatcggc tacgaaaacg acccggtatt ccgtgcgctg 840
gacggctcca ccttcaacct gtcgtccctc ggcgtgcatg acaaggccca cgagtcgacc 900
accgacaaca tcgtcagctt caacgaccac tacgcctcga cgttgtggaa tgtgctgccg 960
ttttccatcg ccaacctgtc gacctgggtg tcgcatttgc cagcgcttac ggcgacggca 1020
tgacgcgtgt gctggaatcg gggttctacg agcaaatgac ccgtgactcg acgattatcg 1080
tcgccaacct gtccgacccg gcgcgcgcca acacctgggt ccaggacctc aaccgcaatg 1140
ccgagccgca cacaggcaat accttcatca tcggcagcga cggcaatgac ctgatccagg 1200
gcggcaaggg cgcggacttc atcgaaggcg gcaagggcaa tgacacgatc cgcgacaaca 1260
gcgggcacaa cacctttttg ttcagcgggc attttggcca ggatcggatt atcggctacc 1320
agccgaccga caggctggtg ttccagggcg ccgacggcag caccgacctg cgcgaccacg 1380
cgaaggccgt gggggccgat acggtgctga gttttggcgc cgactcggtg accctggtcg 1440
gggtcgggct gggcggcctg tggagcgagg gtgtgctgat cagttgaggg cgccgaaagc 1500
gatgaaactg ttgaagaaat tgctgaaatt acagaaaaaa ggcatt 1546
<210> 69
<211> 1546
<212> DNA
<213> Artificial Sequence
<220>
<223> NRC - lipase TliA - NRC
<400> 69
gcgccgaaag cgatgcgtct gttgcgtcgc ttgctgcgtt tacagaaaaa aggcattatg 60
ggtgtatttg actacaagaa cctcggcacc gaagccagca aaaccttgtt cgccgatgcc 120
accgcaatca cgttgtatac ctatcacaac ctggataacg gcttcgcagt cggctaccag 180
caacatggct tggggctcgg cctgccggcc acactggtcg gggcgttgct cggcagcaca 240
gactcccagg gagtgatccc cggcattccc tggaatcctg actcggaaaa ggccgccctg 300
gacgcggtgc acgcagccgg ttggacgcca atcagcgcca gcgcactggg ctacggcggc 360
aaggtggatg cgcggggcac tttttttggc gagaaggccg gctacaccac ggcccaggcc 420
gaagtgctgg gcaagtacga tgacgccggc aaactgctcg agatcggcat cggttttcgt 480
ggcacctcgg gccctcggga aagcctgatt accgactcca tcggcgatct ggtcagcgac 540
ctgctcgccg cgctgggccc caaggactat gcgaaaaact atgccggcga agcgtttggc 600
ggcttgctca agacggtggc cgactatgcc ggcgcccatg gcctgagtgg caaggatgtg 660
ctggtcagcg gccacagcct gggcggcctg gcggtcaaca gcatggccga cctgagcacc 720
agcaaatggg cgggtttcta caaggacgcc aactacctgg cctacgcctc gcccacccag 780
agcgccggcg ataaggtcct gaatatcggc tacgaaaacg acccggtatt ccgtgcgctg 840
gacggctcca ccttcaacct gtcgtccctc ggcgtgcatg acaaggccca cgagtcgacc 900
accgacaaca tcgtcagctt caacgaccac tacgcctcga cgttgtggaa tgtgctgccg 960
ttttccatcg ccaacctgtc gacctgggtg tcgcatttgc cagcgcttac ggcgacggca 1020
tgacgcgtgt gctggaatcg gggttctacg agcaaatgac ccgtgactcg acgattatcg 1080
tcgccaacct gtccgacccg gcgcgcgcca acacctgggt ccaggacctc aaccgcaatg 1140
ccgagccgca cacaggcaat accttcatca tcggcagcga cggcaatgac ctgatccagg 1200
gcggcaaggg cgcggacttc atcgaaggcg gcaagggcaa tgacacgatc cgcgacaaca 1260
gcgggcacaa cacctttttg ttcagcgggc attttggcca ggatcggatt atcggctacc 1320
agccgaccga caggctggtg ttccagggcg ccgacggcag caccgacctg cgcgaccacg 1380
cgaaggccgt gggggccgat acggtgctga gttttggcgc cgactcggtg accctggtcg 1440
gggtcgggct gggcggcctg tggagcgagg gtgtgctgat cagttgaggg cgccgaaagc 1500
gatgcgtctg ttgcgtcgct tgctgcgttt acagaaaaaa ggcatt 1546
<210> 70
<211> 1148
<212> DNA
<213> Artificial Sequence
<220>
<223> Buforin IIb - lipase M37 - Buforin IIb
<400> 70
cgtgctggtc tgcagttccc ggttggtcgt ctgctgcgtc gtctgctgcg tcgtctgctg 60
cgtatgtctt atacaaaaga acaactcatg ttggcattca gctatatgag ctactatggc 120
atcactcaca caggttcagc aaaaaaaaat gctgagctca tccttaaaaa aatgaaagaa 180
gccttgaaaa catggaagcc ttttcaggaa gatgactggg aagtcgtctg gggtcctgcg 240
gtttatacca tgcctttcac aatcttcaac gatgccatga tgtatgtcat acagaagaaa 300
ggtgctgaag gggaatacgt gatagccatt cgcggcacca atccagtatc aatttcagac 360
tggctgttta atgatttcat ggtcagcgca atgaagaagt ggccttacgc atccgttgaa 420
ggccgcatac tcaaaatatc cgaaagtacc agctacggac tgaaaacctt acagaaattg 480
aagccaaaat cccatatccc cggcgaaaat aaaacgattc tgcagttcct gaatgaaaag 540
ataggcccag agggtaaagc aaaaatctgt gtaacaggcc acagtaaagg cggcgccttg 600
tcttccactc tggcactgtg gttgaaggac atccaaggag taaaactctc gcaaaacatc 660
gatatctcaa cgattccgtt tgccggacca acagccggta atgctgactt tgccgattac 720
tttgatgatt gtcttggtga tcaatgcacc cgcattgcca actcgttaga tattgtgcct 780
tatgcctgga ataaaattca ttaaaaaaac ttaaatctat atatatttct gaacaagcat 840
cagttaaacc gcttctatat caacgcgctt taatccgtgc aatgatcgca gaaactaaag 900
gtaaaaaata caagcaaatt aaggcggaaa caccaccatt agaaggcaac attaatccta 960
ttcttattga ataccttgtg caggcagcat atcagcatgt cgtcggttac ccagaattaa 1020
tgggtatgat ggatgatatt cctttaacag acatattcga agatgcgatc gcgggtttgt 1080
tataacgtgc tggtctgcag ttcccggttg gtcgtctgct gcgtcgtctg ctgcgtcgtc 1140
tgctgcgt 1148
<210> 71
<211> 1076
<212> DNA
<213> Artificial Sequence
<220>
<223> B0 - lipase M37 - B0
<400> 71
cgtgctggtc tgcagttccc ggttggtatg tcttatacaa aagaacaact catgttggca 60
ttcagctata tgagctacta tggcatcact cacacaggtt cagcaaaaaa aaatgctgag 120
ctcatcctta aaaaaatgaa agaagccttg aaaacatgga agccttttca ggaagatgac 180
tgggaagtcg tctggggtcc tgcggtttat accatgcctt tcacaatctt caacgatgcc 240
atgatgtatg tcatacagaa gaaaggtgct gaaggggaat acgtgatagc cattcgcggc 300
accaatccag tatcaatttc agactggctg tttaatgatt tcatggtcag cgcaatgaag 360
aagtggcctt acgcatccgt tgaaggccgc atactcaaaa tatccgaaag taccagctac 420
ggactgaaaa ccttacagaa attgaagcca aaatcccata tccccggcga aaataaaacg 480
attctgcagt tcctgaatga aaagataggc ccagagggta aagcaaaaat ctgtgtaaca 540
ggccacagta aaggcggcgc cttgtcttcc actctggcac tgtggttgaa ggacatccaa 600
ggagtaaaac tctcgcaaaa catcgatatc tcaacgattc cgtttgccgg accaacagcc 660
ggtaatgctg actttgccga ttactttgat gattgtcttg gtgatcaatg cacccgcatt 720
gccaactcgt tagatattgt gccttatgcc tggaataaaa ttcattaaaa aaacttaaat 780
ctatatatat ttctgaacaa gcatcagtta aaccgcttct atatcaacgc gctttaatcc 840
gtgcaatgat cgcagaaact aaaggtaaaa aatacaagca aattaaggcg gaaacaccac 900
cattagaagg caacattaat cctattctta ttgaatacct tgtgcaggca gcatatcagc 960
atgtcgtcgg ttacccagaa ttaatgggta tgatggatga tattccttta acagacatat 1020
tcgaagatgc gatcgcgggt ttgttataac gtgctggtct gcagttcccg gttggt 1076
<210> 72
<211> 1136
<212> DNA
<213> Artificial Sequence
<220>
<223> Paracin I - lipase M37 - Paracin I
<400> 72
aaaggaagag gcaaacaggg aggcaaggtg cgtgcgaagg caaagacacg ttcatccatg 60
tcttatacaa aagaacaact catgttggca ttcagctata tgagctacta tggcatcact 120
cacacaggtt cagcaaaaaa aaatgctgag ctcatcctta aaaaaatgaa agaagccttg 180
aaaacatgga agccttttca ggaagatgac tgggaagtcg tctggggtcc tgcggtttat 240
accatgcctt tcacaatctt caacgatgcc atgatgtatg tcatacagaa gaaaggtgct 300
gaaggggaat acgtgatagc cattcgcggc accaatccag tatcaatttc agactggctg 360
tttaatgatt tcatggtcag cgcaatgaag aagtggcctt acgcatccgt tgaaggccgc 420
atactcaaaa tatccgaaag taccagctac ggactgaaaa ccttacagaa attgaagcca 480
aaatcccata tccccggcga aaataaaacg attctgcagt tcctgaatga aaagataggc 540
ccagagggta aagcaaaaat ctgtgtaaca ggccacagta aaggcggcgc cttgtcttcc 600
actctggcac tgtggttgaa ggacatccaa ggagtaaaac tctcgcaaaa catcgatatc 660
tcaacgattc cgtttgccgg accaacagcc ggtaatgctg actttgccga ttactttgat 720
gattgtcttg gtgatcaatg cacccgcatt gccaactcgt tagatattgt gccttatgcc 780
tggaataaaa ttcattaaaa aaacttaaat ctatatatat ttctgaacaa gcatcagtta 840
aaccgcttct atatcaacgc gctttaatcc gtgcaatgat cgcagaaact aaaggtaaaa 900
aatacaagca aattaaggcg gaaacaccac cattagaagg caacattaat cctattctta 960
ttgaatacct tgtgcaggca gcatatcagc atgtcgtcgg ttacccagaa ttaatgggta 1020
tgatggatga tattccttta acagacatat tcgaagatgc gatcgcgggt ttgttataaa 1080
aaggaagagg caaacaggga ggcaaggtgc gtgcgaaggc aaagacacgt tcatcc 1136
<210> 73
<211> 1136
<212> DNA
<213> Artificial Sequence
<220>
<223> NKC - lipase M37 - NKC
<400> 73
gcgccgaaag cgatgaaact gttgaagaaa ttgctgaaat tacagaaaaa aggcattatg 60
tcttatacaa aagaacaact catgttggca ttcagctata tgagctacta tggcatcact 120
cacacaggtt cagcaaaaaa aaatgctgag ctcatcctta aaaaaatgaa agaagccttg 180
aaaacatgga agccttttca ggaagatgac tgggaagtcg tctggggtcc tgcggtttat 240
accatgcctt tcacaatctt caacgatgcc atgatgtatg tcatacagaa gaaaggtgct 300
gaaggggaat acgtgatagc cattcgcggc accaatccag tatcaatttc agactggctg 360
tttaatgatt tcatggtcag cgcaatgaag aagtggcctt acgcatccgt tgaaggccgc 420
atactcaaaa tatccgaaag taccagctac ggactgaaaa ccttacagaa attgaagcca 480
aaatcccata tccccggcga aaataaaacg attctgcagt tcctgaatga aaagataggc 540
ccagagggta aagcaaaaat ctgtgtaaca ggccacagta aaggcggcgc cttgtcttcc 600
actctggcac tgtggttgaa ggacatccaa ggagtaaaac tctcgcaaaa catcgatatc 660
tcaacgattc cgtttgccgg accaacagcc ggtaatgctg actttgccga ttactttgat 720
gattgtcttg gtgatcaatg cacccgcatt gccaactcgt tagatattgt gccttatgcc 780
tggaataaaa ttcattaaaa aaacttaaat ctatatatat ttctgaacaa gcatcagtta 840
aaccgcttct atatcaacgc gctttaatcc gtgcaatgat cgcagaaact aaaggtaaaa 900
aatacaagca aattaaggcg gaaacaccac cattagaagg caacattaat cctattctta 960
ttgaatacct tgtgcaggca gcatatcagc atgtcgtcgg ttacccagaa ttaatgggta 1020
tgatggatga tattccttta acagacatat tcgaagatgc gatcgcgggt ttgttataag 1080
cgccgaaagc gatgaaactg ttgaagaaat tgctgaaatt acagaaaaaa ggcatt 1136
<210> 74
<211> 1136
<212> DNA
<213> Artificial Sequence
<220>
<223> NRC - lipase M37 - NRC
<400> 74
gcgccgaaag cgatgcgtct gttgcgtcgc ttgctgcgtt tacagaaaaa aggcattatg 60
tcttatacaa aagaacaact catgttggca ttcagctata tgagctacta tggcatcact 120
cacacaggtt cagcaaaaaa aaatgctgag ctcatcctta aaaaaatgaa agaagccttg 180
aaaacatgga agccttttca ggaagatgac tgggaagtcg tctggggtcc tgcggtttat 240
accatgcctt tcacaatctt caacgatgcc atgatgtatg tcatacagaa gaaaggtgct 300
gaaggggaat acgtgatagc cattcgcggc accaatccag tatcaatttc agactggctg 360
tttaatgatt tcatggtcag cgcaatgaag aagtggcctt acgcatccgt tgaaggccgc 420
atactcaaaa tatccgaaag taccagctac ggactgaaaa ccttacagaa attgaagcca 480
aaatcccata tccccggcga aaataaaacg attctgcagt tcctgaatga aaagataggc 540
ccagagggta aagcaaaaat ctgtgtaaca ggccacagta aaggcggcgc cttgtcttcc 600
actctggcac tgtggttgaa ggacatccaa ggagtaaaac tctcgcaaaa catcgatatc 660
tcaacgattc cgtttgccgg accaacagcc ggtaatgctg actttgccga ttactttgat 720
gattgtcttg gtgatcaatg cacccgcatt gccaactcgt tagatattgt gccttatgcc 780
tggaataaaa ttcattaaaa aaacttaaat ctatatatat ttctgaacaa gcatcagtta 840
aaccgcttct atatcaacgc gctttaatcc gtgcaatgat cgcagaaact aaaggtaaaa 900
aatacaagca aattaaggcg gaaacaccac cattagaagg caacattaat cctattctta 960
ttgaatacct tgtgcaggca gcatatcagc atgtcgtcgg ttacccagaa ttaatgggta 1020
tgatggatga tattccttta acagacatat tcgaagatgc gatcgcgggt ttgttataag 1080
cgccgaaagc gatgcgtctg ttgcgtcgct tgctgcgttt acagaaaaaa ggcatt 1136
<210> 75
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> primer pET16-L
<400> 75
cgtagaggat cgagatctcg atcc 24
<210> 76
<211> 60
<212> DNA
<213> Artificial Sequence
<220>
<223> primer pET-R(Par)
<400> 76
gccttcgcac gcaccttgcc tccctgtttg cctcttcctt tacgaccttc gatatggccg 60
60
<210> 77
<211> 62
<212> DNA
<213> Artificial Sequence
<220>
<223> primer TliA-rev(Par)
<400> 77
gggaggcaag gtgcgtgcga aggcaaagac acgttcatcc ggtgtatttg actacaagaa 60
cc 62
<210> 78
<211> 35
<212> DNA
<213> Artificial Sequence
<220>
<223> primer NdeI-TliA-for
<400> 78
cttaaggcat atgtcaactg atcagcacac cctcg 35
<210> 79
<211> 60
<212> DNA
<213> Artificial Sequence
<220>
<223> primer pET-R(Buf)
<400> 79
cgacgcagca gacgaccaac cgggaactgc agaccagcac gacgaccttc gatatggccg 60
60
<210> 80
<211> 68
<212> DNA
<213> Artificial Sequence
<220>
<223> primer TliA-rev(Buf)
<400> 80
cccggttggt cgtctgctgc gtcgtctgct gcgtcgtctg ctgcgtggtg tatttgacta 60
caagaacc 68
<210> 81
<211> 44
<212> DNA
<213> Artificial Sequence
<220>
<223> primer pET-R(B0)
<400> 81
caaccgggaa ctgcagacca gcacgacgac cttcgatatg gccg 44
<210> 82
<211> 46
<212> DNA
<213> Artificial Sequence
<220>
<223> primer TliA-rev(B0)
<400> 82
gctggtctgc agttcccggt tggtggtgta tttgactaca agaacc 46
<210> 83
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> primer pET-R(con)
<400> 83
acgaccttcg atatggccg 19
<210> 84
<211> 41
<212> DNA
<213> Artificial Sequence
<220>
<223> primer TliA-rev(con)
<400> 84
cggccatatc gaaggtcgtg gtgtatttga ctacaagaac c 41
<210> 85
<211> 68
<212> DNA
<213> Artificial Sequence
<220>
<223> primer M37-rev(Buf)
<400> 85
cccggttggt cgtctgctgc gtcgtctgct gcgtcgtctg ctgcgtgcat ctccacgcgc 60
caatgatg 68
<210> 86
<211> 35
<212> DNA
<213> Artificial Sequence
<220>
<223> primer Nde-M37-for
<400> 86
cttaaggcat atgttataac aaacccgcga tcgca 35
<210> 87
<211> 62
<212> DNA
<213> Artificial Sequence
<220>
<223> primer M37-rev(Par)
<400> 87
gggaggcaag gtgcgtgcga aggcaaagac acgttcatcc gcatctccac gcgccaatga 60
tg 62
<210> 88
<211> 46
<212> DNA
<213> Artificial Sequence
<220>
<223> primer M37-rev(B0)
<400> 88
gctggtctgc agttcccggt tggtgcatct ccacgcgcca atgatg 46
<210> 89
<211> 62
<212> DNA
<213> Artificial Sequence
<220>
<223> primer M37-rev(NKC)
<400> 89
actgttgaag aaattgctga aattacagaa aaaaggcatt gcatctccac gcgccaatga 60
tg 62
<210> 90
<211> 62
<212> DNA
<213> Artificial Sequence
<220>
<223> primer M37-rev(NRC)
<400> 90
tctgttgcgt cgcttgctgc gtttacagaa aaaaggcatt gcatctccac gcgccaatga 60
tg 62
<210> 91
<211> 42
<212> DNA
<213> Artificial Sequence
<220>
<223> primer M37-rev(con)
<400> 91
gcagcggcca tatcgaaggt cgtgcatctc cacgcgccaa tg 42
Claims (18)
- 양친매성 펩타이드 및 리파아제가 연결된 양친매성 펩타이드-리파아제 결합체를 포함하는, 양친매성 펩타이드 비결합 리파아제 대비, 리파아제의 기질 친화도 개선용 조성물.
- 제1항에 있어서,
상기 양친매성 펩타이드는 부포린 IIb, B0, 파라신 I, NKC 및 NRC로 이루어진 그룹으로부터 선택되는 것인, 조성물.
- 제1항에 있어서, 상기 양친매성 펩타이드는 서열번호 1로 기재된 아미노산 서열로 정의되는 부포린 IIb, 서열번호 2로 기재된 아미노산 서열로 정의되는 B0, 서열번호 3으로 기재된 아미노산 서열로 정의되는 파라신 I, 서열번호 4로 기재된 아미노산 서열로 정의되는 NKC 및 서열번호 5로 기재된 아미노산 서열로 정의되는 NRC로 이루어진 그룹으로부터 선택되는 것인 조성물.
- 제1항에 있어서, 상기 리파아제는 슈도모나스 플루오레슨스(Pseudomonas fluorescens) 유래의 리파아제 TliA(thermostable lipase) 또는 포토박테리움 리포리티쿰(Photobacterium lipolyticum) 유래의 리파아제 M37인 것인 조성물.
- 제4항에 있어서, 상기 TliA는 서열번호 6으로 기재된 아미노산 서열로 정의되는 리파아제이고, M37은 서열번호 7로 기재된 아미노산 서열로 정의되는 리파아제인 것인 조성물.
- 제1항에 있어서, 상기 양친매성 펩타이드는 리파아제의 N-말단 또는 C-말단에 위치하거나, 두 개의 양친매성 펩타이드가 리파아제의 N-말단 및 C-말단 모두에 위치한 것인 조성물.
- 제1항에 있어서, 상기 양친매성 펩타이드와 리파아제는 링커(linker)에 의해 연결된 것인 조성물.
- 제7항에 있어서, 상기 링커는 비펩타이드성 링커 또는 펩타이드성 링커인 것인 조성물.
- 제8항에 있어서, 상기 비펩타이드성 링커는 폴리에틸렌 글리콜 단독 중합체, 폴리프로필렌 글리콜 단독 중합체, 에틸렌 글리콜-프로필렌 글리콜 공중합체, 폴리옥시 에틸화 폴리올, 폴리비닐 알콜, 폴리사카라이드, 덱스트란, 폴리비닐 에틸 에테르, 생분해성 고분자, 지질 중합체, 키틴류, 히아루론산 및 이들의 조합으로 이루어진 군에서 선택된 것인 조성물.
- 제1항에 있어서, 상기 결합체는 서열번호 8 내지 17로 이루어지는 군으로부터 선택되는 아미노산 서열로 정의되는 것인 조성물.
- 제1항 내지 제10항 중 어느 한 항의 조성물에 포함된 양친매성 펩타이드-리파아제 결합체를 코딩하는 폴리뉴클레오티드.
- 제11항에 있어서, 상기 폴리뉴클레오티드는 서열번호 45 내지 74로 이루어지는 군으로부터 선택되는 뉴클레오티드 서열을 포함하는 것인 폴리뉴클레오티드.
- 제11항의 폴리뉴클레오티드를 포함하는 발현벡터.
- 제13항의 발현벡터가 도입된 형질전환체.
- (a) 제14항의 형질전환체를 배양하는 단계; 및
(b) 상기 배양된 형질전환체 또는 이의 배양액으로부터 양친매성 펩타이드-리파아제 결합체를 회수하여 양친매성 펩타이드-리파아제 결합체를 준비하는 단계를 포함하는,
양친매성 펩타이드-리파아제 결합체를 포함하는, 리파아제의 기질 친화성 개선용 조성물의 제조방법.
- 제15항에 있어서, 상기 양친매성 펩타이드-리파아제 결합체는 서열번호 8 내지 17로 이루어지는 군으로부터 선택되는 아미노산 서열로 정의되는 것인 제조방법.
- 제1항 내지 제10항 중 어느 한 항의 조성물과 지질을 반응시키는 단계를 포함하는, 지질의 분해방법.
- 제1항 내지 제10항 중 어느 한 항의 조성물과 유지 및 알코올을 반응시키는 단계를 포함하는, 바이오디젤의 생산방법.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020110079609 | 2011-08-10 | ||
KR20110079609 | 2011-08-10 |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
KR1020120088042A Division KR101677959B1 (ko) | 2011-08-10 | 2012-08-10 | 리파아제 활성이 증대된 양친매성 펩타이드-리파아제 결합체 및 이의 용도 |
Publications (2)
Publication Number | Publication Date |
---|---|
KR20150132813A true KR20150132813A (ko) | 2015-11-26 |
KR101887732B1 KR101887732B1 (ko) | 2018-09-10 |
Family
ID=47669117
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
KR1020120088042A KR101677959B1 (ko) | 2011-08-10 | 2012-08-10 | 리파아제 활성이 증대된 양친매성 펩타이드-리파아제 결합체 및 이의 용도 |
KR1020150155169A KR101887732B1 (ko) | 2011-08-10 | 2015-11-05 | 리파아제 활성이 증대된 양친매성 펩타이드-리파아제 결합체 및 이의 용도 |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
KR1020120088042A KR101677959B1 (ko) | 2011-08-10 | 2012-08-10 | 리파아제 활성이 증대된 양친매성 펩타이드-리파아제 결합체 및 이의 용도 |
Country Status (6)
Country | Link |
---|---|
US (1) | US9527886B2 (ko) |
EP (1) | EP2742069B1 (ko) |
JP (1) | JP5933001B2 (ko) |
KR (2) | KR101677959B1 (ko) |
CN (2) | CN103687880B (ko) |
WO (1) | WO2013022320A2 (ko) |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR102138254B1 (ko) * | 2018-01-22 | 2020-07-29 | 재단법인 지능형 바이오 시스템 설계 및 합성 연구단 | 접착성이 부여된 항균 펩타이드의 제조 방법 및 이의 용도 |
CN113321708B (zh) * | 2021-04-05 | 2022-09-30 | 江苏亢钧生物科技有限公司 | 一种人工设计抗菌肽的制备及其在水产上的应用 |
CN114149987B (zh) * | 2021-12-07 | 2024-02-13 | 安徽大学 | 一种人工改造的β-半乳糖苷酶GaLT1及其在水解乳糖中的应用 |
CN116139248A (zh) * | 2022-11-18 | 2023-05-23 | 中国科学院过程工程研究所 | 一种肽水凝胶、其制备方法及其在治疗中的应用 |
WO2024121058A1 (en) * | 2022-12-05 | 2024-06-13 | Novozymes A/S | A composition comprising a lipase and a peptide |
Family Cites Families (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR100314721B1 (ko) * | 1998-01-22 | 2001-11-23 | 김일웅 | 생물학적 활성이 있는 신규한 펩타이드 |
WO1999048912A1 (en) * | 1998-03-25 | 1999-09-30 | Korea Advanced Institute Of Science And Technology | A novel antimicrobial peptide isolated from parasilurus asotus and its uses |
KR100330136B1 (ko) | 1998-03-25 | 2002-03-27 | 윤덕용 | 메기로부터 분리한 신규한 항균 펩타이드 파라신ⅰ과 그 용도 |
CN1249079C (zh) * | 1999-03-25 | 2006-04-05 | 瓦申泰克尼利南塔基马斯凯斯库斯公司 | 蛋白质的分配方法 |
US6833441B2 (en) * | 2001-08-01 | 2004-12-21 | Abmaxis, Inc. | Compositions and methods for generating chimeric heteromultimers |
WO2005079523A2 (en) * | 2004-02-18 | 2005-09-01 | University Of Georgia Research Foundation, Inc. | Novel teleost derived antimicrobal polypeptides |
KR100836596B1 (ko) * | 2007-01-19 | 2008-06-10 | 한국과학기술원 | 높은 염농도에서 항균 활성을 갖는 펩타이드들 및 이들을포함하는 항균 조성물 |
KR100958095B1 (ko) * | 2007-12-31 | 2010-05-14 | 한국과학기술원 | 번역 동반 시스템을 이용한 항균 펩타이드의 대량 발현방법 |
US8936787B2 (en) | 2008-04-15 | 2015-01-20 | The United States Of America As Represented By The Secretary, Department Of Health And Human Services | Peptides promoting lipid efflux |
WO2011066511A1 (en) | 2009-11-30 | 2011-06-03 | The U.S.A., As Represented By The Secretary Department Of Health And Human Services | Synthetic apoa-1 mimetic amphipathic peptides and methods of use thereof |
CN102120991B (zh) * | 2010-12-21 | 2012-10-10 | 清华大学 | 脂肪酶 |
-
2012
- 2012-08-10 KR KR1020120088042A patent/KR101677959B1/ko active IP Right Grant
- 2012-08-10 CN CN201280029248.XA patent/CN103687880B/zh not_active Expired - Fee Related
- 2012-08-10 JP JP2014523877A patent/JP5933001B2/ja not_active Expired - Fee Related
- 2012-08-10 CN CN201610663163.7A patent/CN106632685A/zh active Pending
- 2012-08-10 EP EP12822946.5A patent/EP2742069B1/en not_active Not-in-force
- 2012-08-10 US US14/123,513 patent/US9527886B2/en not_active Expired - Fee Related
- 2012-08-10 WO PCT/KR2012/006412 patent/WO2013022320A2/en active Application Filing
-
2015
- 2015-11-05 KR KR1020150155169A patent/KR101887732B1/ko active IP Right Grant
Non-Patent Citations (1)
Title |
---|
Journal of Laboratory Automation, Vol. 16, No. 1, Pages 17-37(공개일: 2011. 2.)* * |
Also Published As
Publication number | Publication date |
---|---|
EP2742069A2 (en) | 2014-06-18 |
WO2013022320A3 (en) | 2013-05-02 |
WO2013022320A9 (en) | 2013-05-30 |
CN103687880B (zh) | 2016-09-14 |
EP2742069A4 (en) | 2015-04-01 |
US20140162331A1 (en) | 2014-06-12 |
JP5933001B2 (ja) | 2016-06-08 |
EP2742069B1 (en) | 2018-05-02 |
JP2014524239A (ja) | 2014-09-22 |
KR20130018202A (ko) | 2013-02-20 |
KR101677959B1 (ko) | 2016-11-21 |
US9527886B2 (en) | 2016-12-27 |
CN106632685A (zh) | 2017-05-10 |
WO2013022320A2 (en) | 2013-02-14 |
KR101887732B1 (ko) | 2018-09-10 |
CN103687880A (zh) | 2014-03-26 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
KR101887732B1 (ko) | 리파아제 활성이 증대된 양친매성 펩타이드-리파아제 결합체 및 이의 용도 | |
US20140221608A1 (en) | Acid-cleavable linkers exhibiting altered rates of acid hydrolysis | |
WO2017101801A1 (zh) | 高效的不依赖锌离子的磷脂酶c突变体 | |
US20040142441A1 (en) | Enzymes with lipase/acyltransferase activity, nucleic acids encoding the same and methods of use thereof | |
KR20150136604A (ko) | 삼중 나선 단백질의 정제 | |
KR20190015393A (ko) | 생물 계면활성제 산생 재조합 미생물 | |
CN108251400B (zh) | 脂肪酶及其应用 | |
CN110846301A (zh) | 一种重组几丁质脱乙酰酶及其制备方法和应用 | |
CN110951711B (zh) | 一种具有降解手性酯活性的酯酶及其编码基因和应用 | |
EP3872182A1 (en) | Recombinant microalgae able to produce peptides, polypeptides or proteins of collagen, elastin and their derivatives in the chloroplast of microalgae and associated method thereof | |
KR101383546B1 (ko) | 심해 해저에서 분리된 에스터라제 ktl 4 | |
KR101596435B1 (ko) | 남극 유래 바실러스 푸밀러스 리파아제 | |
KR101383548B1 (ko) | 심해 해저에서 분리된 에스터라제 ktl 9 | |
KR101741873B1 (ko) | 활성이 증진된 에스터라제 융합단백질 | |
RU2818353C2 (ru) | Мутант фосфолипазы С с высокой ферментативной активностью | |
EP4389884A1 (en) | Efficient production of enantiopure d-3-hydroxybutyrate | |
KR102566555B1 (ko) | 델타-9-지방산불포화 효소를 암호화 하는 유전자 및 이의 용도 | |
CN109957028B (zh) | 提高分泌型蛋白的成熟肽产生的方法 | |
KR101438149B1 (ko) | 스타필러코커스 해모라이티쿠스 l62 리파아제의 세포 표면 발현과 트랜스에스테르화 생물 전환 공정의 응용 | |
CN117535253A (zh) | 甲酸酯脱氢酶铁硫亚基及其编码基因、融合蛋白以及它们的应用 | |
KR101383547B1 (ko) | 심해 해저에서 분리된 에스터라제 ktl 7 | |
KR101897189B1 (ko) | 페니실리움 크리소게넘 균주 유래 리파제 및 이를 이용한 리파제의 제조방법 | |
KR20130055507A (ko) | 갯벌 메타게놈 유래 신규 유전자 및 이로부터 얻어지는 포스포리파제 및 리파제 동시 활성을 보이는 신규 단백질 | |
KR101202261B1 (ko) | 심해 해저에서 분리된 리파제 em2l7 | |
KR100997735B1 (ko) | 갯벌 메타게놈 유래 신규 저온성 리파제 및 이의 제조방법 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A107 | Divisional application of patent | ||
A201 | Request for examination | ||
E902 | Notification of reason for refusal | ||
AMND | Amendment | ||
E601 | Decision to refuse application | ||
AMND | Amendment | ||
X701 | Decision to grant (after re-examination) | ||
GRNT | Written decision to grant |