CN115058465A - 一种岩藻糖基化软骨素及其制备方法和应用 - Google Patents
一种岩藻糖基化软骨素及其制备方法和应用 Download PDFInfo
- Publication number
- CN115058465A CN115058465A CN202210763300.XA CN202210763300A CN115058465A CN 115058465 A CN115058465 A CN 115058465A CN 202210763300 A CN202210763300 A CN 202210763300A CN 115058465 A CN115058465 A CN 115058465A
- Authority
- CN
- China
- Prior art keywords
- chondroitin
- glca
- fucosyltransferase
- fucosylated
- galnac
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 229920002567 Chondroitin Polymers 0.000 title claims abstract description 134
- DLGJWSVWTWEWBJ-HGGSSLSASA-N chondroitin Chemical compound CC(O)=N[C@@H]1[C@H](O)O[C@H](CO)[C@H](O)[C@@H]1OC1[C@H](O)[C@H](O)C=C(C(O)=O)O1 DLGJWSVWTWEWBJ-HGGSSLSASA-N 0.000 title claims abstract description 89
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
- 230000033581 fucosylation Effects 0.000 claims abstract description 44
- 238000000034 method Methods 0.000 claims abstract description 40
- 230000004048 modification Effects 0.000 claims abstract description 25
- 238000012986 modification Methods 0.000 claims abstract description 25
- 239000000758 substrate Substances 0.000 claims abstract description 19
- 235000000346 sugar Nutrition 0.000 claims abstract description 15
- 108010019236 Fucosyltransferases Proteins 0.000 claims abstract description 8
- 102000006471 Fucosyltransferases Human genes 0.000 claims abstract description 8
- 229920001542 oligosaccharide Polymers 0.000 claims description 55
- -1 chondroitin oligosaccharide Chemical class 0.000 claims description 51
- 102220470698 BUD13 homolog_V30P_mutation Human genes 0.000 claims description 20
- AEMOLEFTQBMNLQ-AQKNRBDQSA-N D-glucopyranuronic acid Chemical compound OC1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-AQKNRBDQSA-N 0.000 claims description 20
- AEMOLEFTQBMNLQ-UHFFFAOYSA-N beta-D-galactopyranuronic acid Natural products OC1OC(C(O)=O)C(O)C(O)C1O AEMOLEFTQBMNLQ-UHFFFAOYSA-N 0.000 claims description 18
- 239000002773 nucleotide Substances 0.000 claims description 16
- 125000003729 nucleotide group Chemical group 0.000 claims description 16
- 238000002156 mixing Methods 0.000 claims description 15
- 102200044886 rs121913409 Human genes 0.000 claims description 15
- 108010009115 chondroitin synthase Proteins 0.000 claims description 13
- 102000003960 Ligases Human genes 0.000 claims description 12
- 108090000364 Ligases Proteins 0.000 claims description 12
- MBLBDJOUHNCFQT-UHFFFAOYSA-N N-acetyl-D-galactosamine Natural products CC(=O)NC(C=O)C(O)C(O)C(O)CO MBLBDJOUHNCFQT-UHFFFAOYSA-N 0.000 claims description 12
- OVRNDRQMDRJTHS-KEWYIRBNSA-N N-acetyl-D-galactosamine Chemical compound CC(=O)N[C@H]1C(O)O[C@H](CO)[C@H](O)[C@@H]1O OVRNDRQMDRJTHS-KEWYIRBNSA-N 0.000 claims description 11
- LFTYTUAZOPRMMI-NESSUJCYSA-N UDP-N-acetyl-alpha-D-galactosamine Chemical compound O1[C@H](CO)[C@H](O)[C@H](O)[C@@H](NC(=O)C)[C@H]1O[P@](O)(=O)O[P@](O)(=O)OC[C@@H]1[C@@H](O)[C@@H](O)[C@H](N2C(NC(=O)C=C2)=O)O1 LFTYTUAZOPRMMI-NESSUJCYSA-N 0.000 claims description 9
- LFTYTUAZOPRMMI-UHFFFAOYSA-N UNPD164450 Natural products O1C(CO)C(O)C(O)C(NC(=O)C)C1OP(O)(=O)OP(O)(=O)OCC1C(O)C(O)C(N2C(NC(=O)C=C2)=O)O1 LFTYTUAZOPRMMI-UHFFFAOYSA-N 0.000 claims description 7
- 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 6
- 102100039847 Globoside alpha-1,3-N-acetylgalactosaminyltransferase 1 Human genes 0.000 claims description 6
- 101000887519 Homo sapiens Globoside alpha-1,3-N-acetylgalactosaminyltransferase 1 Proteins 0.000 claims description 6
- LQEBEXMHBLQMDB-UHFFFAOYSA-N GDP-L-fucose Natural products OC1C(O)C(O)C(C)OC1OP(O)(=O)OP(O)(=O)OCC1C(O)C(O)C(N2C3=C(C(N=C(N)N3)=O)N=C2)O1 LQEBEXMHBLQMDB-UHFFFAOYSA-N 0.000 claims description 5
- LQEBEXMHBLQMDB-JGQUBWHWSA-N GDP-beta-L-fucose Chemical compound O[C@H]1[C@H](O)[C@H](O)[C@H](C)O[C@@H]1OP(O)(=O)OP(O)(=O)OC[C@@H]1[C@@H](O)[C@@H](O)[C@H](N2C3=C(C(NC(N)=N3)=O)N=C2)O1 LQEBEXMHBLQMDB-JGQUBWHWSA-N 0.000 claims description 5
- 238000000746 purification Methods 0.000 claims description 5
- 150000008163 sugars Chemical class 0.000 claims description 3
- HDYANYHVCAPMJV-UHFFFAOYSA-N Uridine diphospho-D-glucuronic acid Natural products O1C(N2C(NC(=O)C=C2)=O)C(O)C(O)C1COP(O)(=O)OP(O)(=O)OC1OC(C(O)=O)C(O)C(O)C1O HDYANYHVCAPMJV-UHFFFAOYSA-N 0.000 claims description 2
- 238000006911 enzymatic reaction Methods 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 3
- WMWKTCPGFOEPBD-YGIWDPDDSA-N azane;(2s,3s,4s,5r,6r)-6-[[[(2r,3s,4r,5r)-5-(2,4-dioxopyrimidin-1-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-hydroxyphosphoryl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound N.C([C@@H]1[C@H]([C@H]([C@@H](O1)N1C(NC(=O)C=C1)=O)O)O)OP(O)(=O)OP(O)(=O)O[C@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O WMWKTCPGFOEPBD-YGIWDPDDSA-N 0.000 claims 1
- 102000004190 Enzymes Human genes 0.000 abstract description 9
- 108090000790 Enzymes Proteins 0.000 abstract description 9
- SQDAZGGFXASXDW-UHFFFAOYSA-N 5-bromo-2-(trifluoromethoxy)pyridine Chemical compound FC(F)(F)OC1=CC=C(Br)C=N1 SQDAZGGFXASXDW-UHFFFAOYSA-N 0.000 abstract description 6
- 229920001287 Chondroitin sulfate Polymers 0.000 abstract description 6
- 229940059329 chondroitin sulfate Drugs 0.000 abstract description 6
- 239000003146 anticoagulant agent Substances 0.000 abstract description 5
- 230000002194 synthesizing effect Effects 0.000 abstract description 4
- 238000009776 industrial production Methods 0.000 abstract description 3
- 238000006555 catalytic reaction Methods 0.000 abstract description 2
- 150000001720 carbohydrates Chemical class 0.000 abstract 1
- 238000006243 chemical reaction Methods 0.000 description 95
- LQEBEXMHBLQMDB-QIXZNPMTSA-N GDP-L-fucose Chemical compound O[C@H]1[C@H](O)[C@H](O)[C@H](C)OC1OP(O)(=O)OP(O)(=O)OC[C@@H]1[C@@H](O)[C@@H](O)[C@H](N2C3=C(C(NC(N)=N3)=O)N=C2)O1 LQEBEXMHBLQMDB-QIXZNPMTSA-N 0.000 description 38
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 36
- 239000002808 molecular sieve Substances 0.000 description 32
- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 description 32
- 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 27
- 239000003957 anion exchange resin Substances 0.000 description 26
- 238000011033 desalting Methods 0.000 description 24
- 239000000047 product Substances 0.000 description 20
- 239000000243 solution Substances 0.000 description 20
- 238000001514 detection method Methods 0.000 description 16
- SHZGCJCMOBCMKK-DHVFOXMCSA-N L-fucopyranose Chemical group C[C@@H]1OC(O)[C@@H](O)[C@H](O)[C@@H]1O SHZGCJCMOBCMKK-DHVFOXMCSA-N 0.000 description 15
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 14
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 12
- 239000006228 supernatant Substances 0.000 description 12
- 238000003786 synthesis reaction Methods 0.000 description 11
- 239000007864 aqueous solution Substances 0.000 description 9
- 230000015572 biosynthetic process Effects 0.000 description 9
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 8
- HDYANYHVCAPMJV-LXQIFKJMSA-N UDP-alpha-D-glucuronic acid Chemical compound C([C@@H]1[C@H]([C@H]([C@@H](O1)N1C(NC(=O)C=C1)=O)O)O)OP(O)(=O)OP(O)(=O)O[C@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O HDYANYHVCAPMJV-LXQIFKJMSA-N 0.000 description 8
- PNNNRSAQSRJVSB-SLPGGIOYSA-N Fucose Natural products C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)C=O PNNNRSAQSRJVSB-SLPGGIOYSA-N 0.000 description 7
- 238000001035 drying Methods 0.000 description 7
- 150000001875 compounds Chemical class 0.000 description 6
- 150000002482 oligosaccharides Chemical class 0.000 description 6
- 238000000926 separation method Methods 0.000 description 6
- PZUPAGRIHCRVKN-UHFFFAOYSA-N 5-[5-[3,4-dihydroxy-6-[(3,4,5-trihydroxyoxan-2-yl)oxymethyl]-5-[3,4,5-trihydroxy-6-[(3,4,5-trihydroxyoxan-2-yl)oxymethyl]oxan-2-yl]oxyoxan-2-yl]oxy-3,4-dihydroxy-6-[(3,4,5-trihydroxyoxan-2-yl)oxymethyl]oxan-2-yl]oxy-6-(hydroxymethyl)oxane-2,3,4-triol Chemical compound OCC1OC(O)C(O)C(O)C1OC1C(O)C(O)C(OC2C(C(O)C(OC3C(C(O)C(O)C(COC4C(C(O)C(O)CO4)O)O3)O)C(COC3C(C(O)C(O)CO3)O)O2)O)C(COC2C(C(O)C(O)CO2)O)O1 PZUPAGRIHCRVKN-UHFFFAOYSA-N 0.000 description 5
- 241000606124 Bacteroides fragilis Species 0.000 description 5
- 241000590002 Helicobacter pylori Species 0.000 description 5
- 239000007853 buffer solution Substances 0.000 description 5
- 229940037467 helicobacter pylori Drugs 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 4
- 150000001413 amino acids Chemical group 0.000 description 4
- 108010062796 arginyllysine Proteins 0.000 description 4
- 230000003197 catalytic effect Effects 0.000 description 4
- 239000013598 vector Substances 0.000 description 4
- 241000588724 Escherichia coli Species 0.000 description 3
- 241000251511 Holothuroidea Species 0.000 description 3
- 229940127219 anticoagulant drug Drugs 0.000 description 3
- 239000008194 pharmaceutical composition Substances 0.000 description 3
- 229920001282 polysaccharide Polymers 0.000 description 3
- 239000005017 polysaccharide Substances 0.000 description 3
- 238000000108 ultra-filtration Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- DVJSJDDYCYSMFR-ZKWXMUAHSA-N Ala-Ile-Gly Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(O)=O DVJSJDDYCYSMFR-ZKWXMUAHSA-N 0.000 description 2
- BYLSYQASFJJBCL-DCAQKATOSA-N Asn-Pro-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O BYLSYQASFJJBCL-DCAQKATOSA-N 0.000 description 2
- BLQBMRNMBAYREH-UWJYBYFXSA-N Asp-Ala-Tyr Chemical compound N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O BLQBMRNMBAYREH-UWJYBYFXSA-N 0.000 description 2
- 241001135228 Bacteroides ovatus Species 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- VCUNGPMMPNJSGS-JYJNAYRXSA-N Gln-Tyr-Lys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)N)N)O VCUNGPMMPNJSGS-JYJNAYRXSA-N 0.000 description 2
- 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 2
- 241000589989 Helicobacter Species 0.000 description 2
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 2
- WGVPDSNCHDEDBP-KKUMJFAQSA-N His-Asp-Phe Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O WGVPDSNCHDEDBP-KKUMJFAQSA-N 0.000 description 2
- 241000282414 Homo sapiens Species 0.000 description 2
- FADYJNXDPBKVCA-UHFFFAOYSA-N L-Phenylalanyl-L-lysin Natural products NCCCCC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FADYJNXDPBKVCA-UHFFFAOYSA-N 0.000 description 2
- LHSGPCFBGJHPCY-UHFFFAOYSA-N L-leucine-L-tyrosine Natural products CC(C)CC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 LHSGPCFBGJHPCY-UHFFFAOYSA-N 0.000 description 2
- 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 2
- OVRNDRQMDRJTHS-CBQIKETKSA-N N-Acetyl-D-Galactosamine Chemical compound CC(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@H](O)[C@@H]1O OVRNDRQMDRJTHS-CBQIKETKSA-N 0.000 description 2
- XZFYRXDAULDNFX-UHFFFAOYSA-N N-L-cysteinyl-L-phenylalanine Natural products SCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XZFYRXDAULDNFX-UHFFFAOYSA-N 0.000 description 2
- XMBSYZWANAQXEV-UHFFFAOYSA-N N-alpha-L-glutamyl-L-phenylalanine Natural products OC(=O)CCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XMBSYZWANAQXEV-UHFFFAOYSA-N 0.000 description 2
- 241000606856 Pasteurella multocida Species 0.000 description 2
- 208000007536 Thrombosis Diseases 0.000 description 2
- 241000607626 Vibrio cholerae Species 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 108010077245 asparaginyl-proline Proteins 0.000 description 2
- 108010047857 aspartylglycine Proteins 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 150000002016 disaccharides Chemical class 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 108010001671 galactoside 3-fucosyltransferase Proteins 0.000 description 2
- 108010081551 glycylphenylalanine Proteins 0.000 description 2
- 229960002897 heparin Drugs 0.000 description 2
- 229920000669 heparin Polymers 0.000 description 2
- 239000003456 ion exchange resin Substances 0.000 description 2
- 229920003303 ion-exchange polymer Polymers 0.000 description 2
- 239000003055 low molecular weight heparin Substances 0.000 description 2
- 229910001629 magnesium chloride Inorganic materials 0.000 description 2
- 238000001728 nano-filtration Methods 0.000 description 2
- 229940051027 pasteurella multocida Drugs 0.000 description 2
- 239000013600 plasmid vector Substances 0.000 description 2
- 229920002401 polyacrylamide Polymers 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 108010026333 seryl-proline Proteins 0.000 description 2
- 238000005556 structure-activity relationship Methods 0.000 description 2
- 108010078580 tyrosylleucine Proteins 0.000 description 2
- 239000013603 viral vector Substances 0.000 description 2
- JNTMAZFVYNDPLB-PEDHHIEDSA-N (2S,3S)-2-[[[(2S)-1-[(2S,3S)-2-amino-3-methyl-1-oxopentyl]-2-pyrrolidinyl]-oxomethyl]amino]-3-methylpentanoic acid Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JNTMAZFVYNDPLB-PEDHHIEDSA-N 0.000 description 1
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 1
- 241000606791 Actinobacillus ureae Species 0.000 description 1
- 241000186361 Actinobacteria <class> Species 0.000 description 1
- 241000186046 Actinomyces Species 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 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 1
- 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 1
- LDLSENBXQNDTPB-DCAQKATOSA-N Ala-Lys-Arg Chemical compound NCCCC[C@H](NC(=O)[C@@H](N)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N LDLSENBXQNDTPB-DCAQKATOSA-N 0.000 description 1
- SDZRIBWEVVRDQI-CIUDSAMLSA-N Ala-Lys-Asp Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(O)=O SDZRIBWEVVRDQI-CIUDSAMLSA-N 0.000 description 1
- KQESEZXHYOUIIM-CQDKDKBSSA-N Ala-Lys-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O KQESEZXHYOUIIM-CQDKDKBSSA-N 0.000 description 1
- CNQAFFMNJIQYGX-DRZSPHRISA-N Ala-Phe-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C)CC1=CC=CC=C1 CNQAFFMNJIQYGX-DRZSPHRISA-N 0.000 description 1
- JAQNUEWEJWBVAY-WBAXXEDZSA-N Ala-Phe-Phe Chemical compound C([C@H](NC(=O)[C@@H](N)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 JAQNUEWEJWBVAY-WBAXXEDZSA-N 0.000 description 1
- CQJHFKKGZXKZBC-BPNCWPANSA-N Ala-Pro-Tyr Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 CQJHFKKGZXKZBC-BPNCWPANSA-N 0.000 description 1
- DYXOFPBJBAHWFY-JBDRJPRFSA-N Ala-Ser-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](C)N DYXOFPBJBAHWFY-JBDRJPRFSA-N 0.000 description 1
- PEEYDECOOVQKRZ-DLOVCJGASA-N Ala-Ser-Phe Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O PEEYDECOOVQKRZ-DLOVCJGASA-N 0.000 description 1
- 101710098620 Alpha-1,2-fucosyltransferase Proteins 0.000 description 1
- ZTKHZAXGTFXUDD-VEVYYDQMSA-N Arg-Asn-Thr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O ZTKHZAXGTFXUDD-VEVYYDQMSA-N 0.000 description 1
- HAVKMRGWNXMCDR-STQMWFEESA-N Arg-Gly-Phe Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O HAVKMRGWNXMCDR-STQMWFEESA-N 0.000 description 1
- NMRHDSAOIURTNT-RWMBFGLXSA-N Arg-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N NMRHDSAOIURTNT-RWMBFGLXSA-N 0.000 description 1
- UVTGNSWSRSCPLP-UHFFFAOYSA-N Arg-Tyr Natural products NC(CCNC(=N)N)C(=O)NC(Cc1ccc(O)cc1)C(=O)O UVTGNSWSRSCPLP-UHFFFAOYSA-N 0.000 description 1
- QQEWINYJRFBLNN-DLOVCJGASA-N Asn-Ala-Phe Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 QQEWINYJRFBLNN-DLOVCJGASA-N 0.000 description 1
- WVCJSDCHTUTONA-FXQIFTODSA-N Asn-Asp-Arg Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O WVCJSDCHTUTONA-FXQIFTODSA-N 0.000 description 1
- LVHMEJJWEXBMKK-GMOBBJLQSA-N Asn-Ile-Met Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)O)NC(=O)[C@H](CC(=O)N)N LVHMEJJWEXBMKK-GMOBBJLQSA-N 0.000 description 1
- HFPXZWPUVFVNLL-GUBZILKMSA-N Asn-Leu-Gln Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O HFPXZWPUVFVNLL-GUBZILKMSA-N 0.000 description 1
- RCFGLXMZDYNRSC-CIUDSAMLSA-N Asn-Lys-Ala Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O RCFGLXMZDYNRSC-CIUDSAMLSA-N 0.000 description 1
- ORJQQZIXTOYGGH-SRVKXCTJSA-N Asn-Lys-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O ORJQQZIXTOYGGH-SRVKXCTJSA-N 0.000 description 1
- ZYPWIUFLYMQZBS-SRVKXCTJSA-N Asn-Lys-Lys Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)N)N ZYPWIUFLYMQZBS-SRVKXCTJSA-N 0.000 description 1
- RTFWCVDISAMGEQ-SRVKXCTJSA-N Asn-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](CC(=O)N)N RTFWCVDISAMGEQ-SRVKXCTJSA-N 0.000 description 1
- JTXVXGXTRXMOFJ-FXQIFTODSA-N Asn-Pro-Asn Chemical compound NC(=O)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(O)=O JTXVXGXTRXMOFJ-FXQIFTODSA-N 0.000 description 1
- OOXUBGLNDRGOKT-FXQIFTODSA-N Asn-Ser-Arg Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O OOXUBGLNDRGOKT-FXQIFTODSA-N 0.000 description 1
- 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 1
- JBDLMLZNDRLDIX-HJGDQZAQSA-N Asn-Thr-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O JBDLMLZNDRLDIX-HJGDQZAQSA-N 0.000 description 1
- 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 1
- LMIWYCWRJVMAIQ-NHCYSSNCSA-N Asn-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)N)N LMIWYCWRJVMAIQ-NHCYSSNCSA-N 0.000 description 1
- PBVLJOIPOGUQQP-CIUDSAMLSA-N Asp-Ala-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O PBVLJOIPOGUQQP-CIUDSAMLSA-N 0.000 description 1
- ILJQISGMGXRZQQ-IHRRRGAJSA-N Asp-Arg-Tyr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ILJQISGMGXRZQQ-IHRRRGAJSA-N 0.000 description 1
- GHODABZPVZMWCE-FXQIFTODSA-N Asp-Glu-Glu Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O GHODABZPVZMWCE-FXQIFTODSA-N 0.000 description 1
- DGKCOYGQLNWNCJ-ACZMJKKPSA-N Asp-Glu-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O DGKCOYGQLNWNCJ-ACZMJKKPSA-N 0.000 description 1
- PZXPWHFYZXTFBI-YUMQZZPRSA-N Asp-Gly-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC(O)=O PZXPWHFYZXTFBI-YUMQZZPRSA-N 0.000 description 1
- 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 1
- QNMKWNONJGKJJC-NHCYSSNCSA-N Asp-Leu-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O QNMKWNONJGKJJC-NHCYSSNCSA-N 0.000 description 1
- IDDMGSKZQDEDGA-SRVKXCTJSA-N Asp-Phe-Asn Chemical compound OC(=O)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CC(N)=O)C(O)=O)CC1=CC=CC=C1 IDDMGSKZQDEDGA-SRVKXCTJSA-N 0.000 description 1
- MVRGBQGZSDJBSM-GMOBBJLQSA-N Asp-Pro-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](CC(=O)O)N MVRGBQGZSDJBSM-GMOBBJLQSA-N 0.000 description 1
- KGHLGJAXYSVNJP-WHFBIAKZSA-N Asp-Ser-Gly Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O KGHLGJAXYSVNJP-WHFBIAKZSA-N 0.000 description 1
- QSFHZPQUAAQHAQ-CIUDSAMLSA-N Asp-Ser-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O QSFHZPQUAAQHAQ-CIUDSAMLSA-N 0.000 description 1
- ZQFRDAZBTSFGGW-SRVKXCTJSA-N Asp-Ser-Phe Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O ZQFRDAZBTSFGGW-SRVKXCTJSA-N 0.000 description 1
- MGSVBZIBCCKGCY-ZLUOBGJFSA-N Asp-Ser-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O MGSVBZIBCCKGCY-ZLUOBGJFSA-N 0.000 description 1
- RSMZEHCMIOKNMW-GSSVUCPTSA-N Asp-Thr-Thr 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)O)C(O)=O RSMZEHCMIOKNMW-GSSVUCPTSA-N 0.000 description 1
- FIRWLDUOFOULCA-XIRDDKMYSA-N Asp-Trp-Lys Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)O)N FIRWLDUOFOULCA-XIRDDKMYSA-N 0.000 description 1
- OTKUAVXGMREHRX-CFMVVWHZSA-N Asp-Tyr-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC(O)=O)CC1=CC=C(O)C=C1 OTKUAVXGMREHRX-CFMVVWHZSA-N 0.000 description 1
- 241000228212 Aspergillus Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241000606125 Bacteroides Species 0.000 description 1
- GMXSSZUVDNPRMA-FXQIFTODSA-N Cys-Arg-Asp Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(O)=O GMXSSZUVDNPRMA-FXQIFTODSA-N 0.000 description 1
- XRTISHJEPHMBJG-SRVKXCTJSA-N Cys-Asp-Tyr Chemical compound SC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 XRTISHJEPHMBJG-SRVKXCTJSA-N 0.000 description 1
- FIADUEYFRSCCIK-CIUDSAMLSA-N Cys-Glu-Arg Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O FIADUEYFRSCCIK-CIUDSAMLSA-N 0.000 description 1
- SRIRHERUAMYIOQ-CIUDSAMLSA-N Cys-Leu-Ser Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O SRIRHERUAMYIOQ-CIUDSAMLSA-N 0.000 description 1
- OZSBRCONEMXYOJ-AVGNSLFASA-N Cys-Phe-Glu Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CS)N OZSBRCONEMXYOJ-AVGNSLFASA-N 0.000 description 1
- 108010090461 DFG peptide Proteins 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 description 1
- ZFADFBPRMSBPOT-KKUMJFAQSA-N Gln-Arg-Phe Chemical compound N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](Cc1ccccc1)C(O)=O ZFADFBPRMSBPOT-KKUMJFAQSA-N 0.000 description 1
- AAOBFSKXAVIORT-GUBZILKMSA-N Gln-Asn-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O AAOBFSKXAVIORT-GUBZILKMSA-N 0.000 description 1
- CKNUKHBRCSMKMO-XHNCKOQMSA-N Gln-Asn-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)N)NC(=O)[C@H](CCC(=O)N)N)C(=O)O CKNUKHBRCSMKMO-XHNCKOQMSA-N 0.000 description 1
- ICRKQMRFXYDYMK-LAEOZQHASA-N Gln-Val-Asn Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O ICRKQMRFXYDYMK-LAEOZQHASA-N 0.000 description 1
- RLZBLVSJDFHDBL-KBIXCLLPSA-N Glu-Ala-Ile Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O RLZBLVSJDFHDBL-KBIXCLLPSA-N 0.000 description 1
- KKCUFHUTMKQQCF-SRVKXCTJSA-N Glu-Arg-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O KKCUFHUTMKQQCF-SRVKXCTJSA-N 0.000 description 1
- 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 1
- FGSGPLRPQCZBSQ-AVGNSLFASA-N Glu-Phe-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O FGSGPLRPQCZBSQ-AVGNSLFASA-N 0.000 description 1
- UDEPRBFQTWGLCW-CIUDSAMLSA-N Glu-Pro-Asp Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(O)=O UDEPRBFQTWGLCW-CIUDSAMLSA-N 0.000 description 1
- ZKONLKQGTNVAPR-DCAQKATOSA-N Glu-Pro-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](CCC(=O)O)N ZKONLKQGTNVAPR-DCAQKATOSA-N 0.000 description 1
- HMJULNMJWOZNFI-XHNCKOQMSA-N Glu-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)[C@H](CCC(=O)O)N)C(=O)O HMJULNMJWOZNFI-XHNCKOQMSA-N 0.000 description 1
- CQGBSALYGOXQPE-HTUGSXCWSA-N Glu-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](CCC(=O)O)N)O CQGBSALYGOXQPE-HTUGSXCWSA-N 0.000 description 1
- HJTSRYLPAYGEEC-SIUGBPQLSA-N Glu-Tyr-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](CCC(=O)O)N HJTSRYLPAYGEEC-SIUGBPQLSA-N 0.000 description 1
- JXYMPBCYRKWJEE-BQBZGAKWSA-N Gly-Arg-Ala Chemical compound [H]NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(O)=O JXYMPBCYRKWJEE-BQBZGAKWSA-N 0.000 description 1
- HDNXXTBKOJKWNN-WDSKDSINSA-N Gly-Glu-Asn Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O HDNXXTBKOJKWNN-WDSKDSINSA-N 0.000 description 1
- GAFKBWKVXNERFA-QWRGUYRKSA-N Gly-Phe-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CC=CC=C1 GAFKBWKVXNERFA-QWRGUYRKSA-N 0.000 description 1
- GGAPHLIUUTVYMX-QWRGUYRKSA-N Gly-Phe-Ser Chemical compound OC[C@@H](C([O-])=O)NC(=O)[C@@H](NC(=O)C[NH3+])CC1=CC=CC=C1 GGAPHLIUUTVYMX-QWRGUYRKSA-N 0.000 description 1
- ZLCLYFGMKFCDCN-XPUUQOCRSA-N Gly-Ser-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CO)NC(=O)CN)C(O)=O ZLCLYFGMKFCDCN-XPUUQOCRSA-N 0.000 description 1
- 241001503513 Helicobacter bilis Species 0.000 description 1
- WZOGEMJIZBNFBK-CIUDSAMLSA-N His-Asp-Asn Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O WZOGEMJIZBNFBK-CIUDSAMLSA-N 0.000 description 1
- GNBHSMFBUNEWCJ-DCAQKATOSA-N His-Pro-Asn Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(O)=O GNBHSMFBUNEWCJ-DCAQKATOSA-N 0.000 description 1
- IXQGOKWTQPCIQM-YJRXYDGGSA-N His-Thr-His Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CC2=CN=CN2)N)O IXQGOKWTQPCIQM-YJRXYDGGSA-N 0.000 description 1
- HGNUKGZQASSBKQ-PCBIJLKTSA-N Ile-Asp-Phe Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N HGNUKGZQASSBKQ-PCBIJLKTSA-N 0.000 description 1
- LLZLRXBTOOFODM-QSFUFRPTSA-N Ile-Asp-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](C(C)C)C(=O)O)N LLZLRXBTOOFODM-QSFUFRPTSA-N 0.000 description 1
- LPXHYGGZJOCAFR-MNXVOIDGSA-N Ile-Glu-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC(C)C)C(=O)O)N LPXHYGGZJOCAFR-MNXVOIDGSA-N 0.000 description 1
- XLCZWMJPVGRWHJ-KQXIARHKSA-N Ile-Glu-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N1CCC[C@@H]1C(=O)O)N XLCZWMJPVGRWHJ-KQXIARHKSA-N 0.000 description 1
- 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 1
- CDGLBYSAZFIIJO-RCOVLWMOSA-N Ile-Gly-Gly Chemical compound CC[C@H](C)[C@H]([NH3+])C(=O)NCC(=O)NCC([O-])=O CDGLBYSAZFIIJO-RCOVLWMOSA-N 0.000 description 1
- TWPSALMCEHCIOY-YTFOTSKYSA-N Ile-Ile-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(=O)O)N TWPSALMCEHCIOY-YTFOTSKYSA-N 0.000 description 1
- GVKKVHNRTUFCCE-BJDJZHNGSA-N Ile-Leu-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)O)N GVKKVHNRTUFCCE-BJDJZHNGSA-N 0.000 description 1
- ADDYYRVQQZFIMW-MNXVOIDGSA-N Ile-Lys-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N ADDYYRVQQZFIMW-MNXVOIDGSA-N 0.000 description 1
- FFAUOCITXBMRBT-YTFOTSKYSA-N Ile-Lys-Ile Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O FFAUOCITXBMRBT-YTFOTSKYSA-N 0.000 description 1
- MLSUZXHSNRBDCI-CYDGBPFRSA-N Ile-Pro-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(=O)O)N MLSUZXHSNRBDCI-CYDGBPFRSA-N 0.000 description 1
- OMDWJWGZGMCQND-CFMVVWHZSA-N Ile-Tyr-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC(=O)O)C(=O)O)N OMDWJWGZGMCQND-CFMVVWHZSA-N 0.000 description 1
- 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 1
- YHFPHRUWZMEOIX-CYDGBPFRSA-N Ile-Val-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(=O)O)N YHFPHRUWZMEOIX-CYDGBPFRSA-N 0.000 description 1
- 108010065920 Insulin Lispro Proteins 0.000 description 1
- 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 1
- 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 1
- 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 1
- OGCQGUIWMSBHRZ-CIUDSAMLSA-N Leu-Asn-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O OGCQGUIWMSBHRZ-CIUDSAMLSA-N 0.000 description 1
- YKNBJXOJTURHCU-DCAQKATOSA-N Leu-Asp-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N YKNBJXOJTURHCU-DCAQKATOSA-N 0.000 description 1
- MYGQXVYRZMKRDB-SRVKXCTJSA-N Leu-Asp-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCCCN MYGQXVYRZMKRDB-SRVKXCTJSA-N 0.000 description 1
- XVSJMWYYLHPDKY-DCAQKATOSA-N Leu-Asp-Met Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCSC)C(O)=O XVSJMWYYLHPDKY-DCAQKATOSA-N 0.000 description 1
- JQSXWJXBASFONF-KKUMJFAQSA-N Leu-Asp-Phe Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O JQSXWJXBASFONF-KKUMJFAQSA-N 0.000 description 1
- IIKJNQWOQIWWMR-CIUDSAMLSA-N Leu-Cys-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CC(C)C)N IIKJNQWOQIWWMR-CIUDSAMLSA-N 0.000 description 1
- VPKIQULSKFVCSM-SRVKXCTJSA-N Leu-Gln-Arg Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O VPKIQULSKFVCSM-SRVKXCTJSA-N 0.000 description 1
- HQUXQAMSWFIRET-AVGNSLFASA-N Leu-Glu-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CCCCN HQUXQAMSWFIRET-AVGNSLFASA-N 0.000 description 1
- AOFYPTOHESIBFZ-KKUMJFAQSA-N Leu-His-His Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](Cc1cnc[nH]1)C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O AOFYPTOHESIBFZ-KKUMJFAQSA-N 0.000 description 1
- IAJFFZORSWOZPQ-SRVKXCTJSA-N Leu-Leu-Asn Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O IAJFFZORSWOZPQ-SRVKXCTJSA-N 0.000 description 1
- 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 1
- 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 1
- ZAVCJRJOQKIOJW-KKUMJFAQSA-N Leu-Phe-Asp Chemical compound CC(C)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CC(O)=O)C(O)=O)CC1=CC=CC=C1 ZAVCJRJOQKIOJW-KKUMJFAQSA-N 0.000 description 1
- SIGZKCWZEBFNAK-QAETUUGQSA-N Leu-Ser-Ser-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 SIGZKCWZEBFNAK-QAETUUGQSA-N 0.000 description 1
- RIHIGSWBLHSGLV-CQDKDKBSSA-N Leu-Tyr-Ala 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](C)C(O)=O RIHIGSWBLHSGLV-CQDKDKBSSA-N 0.000 description 1
- VJGQRELPQWNURN-JYJNAYRXSA-N Leu-Tyr-Glu 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(O)=O)C(O)=O VJGQRELPQWNURN-JYJNAYRXSA-N 0.000 description 1
- XZNJZXJZBMBGGS-NHCYSSNCSA-N Leu-Val-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O XZNJZXJZBMBGGS-NHCYSSNCSA-N 0.000 description 1
- YQFZRHYZLARWDY-IHRRRGAJSA-N Leu-Val-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCCN YQFZRHYZLARWDY-IHRRRGAJSA-N 0.000 description 1
- 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 1
- 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 1
- QYOXSYXPHUHOJR-GUBZILKMSA-N Lys-Asn-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O QYOXSYXPHUHOJR-GUBZILKMSA-N 0.000 description 1
- YVSHZSUKQHNDHD-KKUMJFAQSA-N Lys-Asn-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CCCCN)N YVSHZSUKQHNDHD-KKUMJFAQSA-N 0.000 description 1
- LMVOVCYVZBBWQB-SRVKXCTJSA-N Lys-Asp-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCCCN LMVOVCYVZBBWQB-SRVKXCTJSA-N 0.000 description 1
- IMAKMJCBYCSMHM-AVGNSLFASA-N Lys-Glu-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CCCCN IMAKMJCBYCSMHM-AVGNSLFASA-N 0.000 description 1
- JYXBNQOKPRQNQS-YTFOTSKYSA-N Lys-Ile-Ile Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JYXBNQOKPRQNQS-YTFOTSKYSA-N 0.000 description 1
- ZXFRGTAIIZHNHG-AJNGGQMLSA-N Lys-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](CCCCN)N ZXFRGTAIIZHNHG-AJNGGQMLSA-N 0.000 description 1
- CBNMHRCLYBJIIZ-XUXIUFHCSA-N Lys-Ile-Met Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)O)NC(=O)[C@H](CCCCN)N CBNMHRCLYBJIIZ-XUXIUFHCSA-N 0.000 description 1
- YXPJCVNIDDKGOE-MELADBBJSA-N Lys-Lys-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCCN)NC(=O)[C@H](CCCCN)N)C(=O)O YXPJCVNIDDKGOE-MELADBBJSA-N 0.000 description 1
- QQPSCXKFDSORFT-IHRRRGAJSA-N Lys-Lys-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CCCCN QQPSCXKFDSORFT-IHRRRGAJSA-N 0.000 description 1
- URGPVYGVWLIRGT-DCAQKATOSA-N Lys-Met-Ala Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](C)C(O)=O URGPVYGVWLIRGT-DCAQKATOSA-N 0.000 description 1
- AZOFEHCPMBRNFD-BZSNNMDCSA-N Lys-Phe-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CCCCN)C(O)=O)CC1=CC=CC=C1 AZOFEHCPMBRNFD-BZSNNMDCSA-N 0.000 description 1
- SQXZLVXQXWILKW-KKUMJFAQSA-N Lys-Ser-Phe Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O SQXZLVXQXWILKW-KKUMJFAQSA-N 0.000 description 1
- 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 1
- MIMXMVDLMDMOJD-BZSNNMDCSA-N Lys-Tyr-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(O)=O MIMXMVDLMDMOJD-BZSNNMDCSA-N 0.000 description 1
- HGKJFNCLOHKEHS-FXQIFTODSA-N Met-Cys-Asp Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CS)C(=O)N[C@H](C(O)=O)CC(O)=O HGKJFNCLOHKEHS-FXQIFTODSA-N 0.000 description 1
- 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 1
- CGUYGMFQZCYJSG-DCAQKATOSA-N Met-Lys-Ser Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O CGUYGMFQZCYJSG-DCAQKATOSA-N 0.000 description 1
- KBTQZYASLSUFJR-KKUMJFAQSA-N Met-Phe-Gln Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N KBTQZYASLSUFJR-KKUMJFAQSA-N 0.000 description 1
- RSOMVHWMIAZNLE-HJWJTTGWSA-N Met-Phe-Ile Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O RSOMVHWMIAZNLE-HJWJTTGWSA-N 0.000 description 1
- YLDSJJOGQNEQJK-AVGNSLFASA-N Met-Pro-Leu Chemical compound CSCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O YLDSJJOGQNEQJK-AVGNSLFASA-N 0.000 description 1
- 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 1
- 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 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- KIEPQOIQHFKQLK-PCBIJLKTSA-N Phe-Asn-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O KIEPQOIQHFKQLK-PCBIJLKTSA-N 0.000 description 1
- MECSIDWUTYRHRJ-KKUMJFAQSA-N Phe-Asn-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O MECSIDWUTYRHRJ-KKUMJFAQSA-N 0.000 description 1
- WGXOKDLDIWSOCV-MELADBBJSA-N Phe-Asn-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC2=CC=CC=C2)N)C(=O)O WGXOKDLDIWSOCV-MELADBBJSA-N 0.000 description 1
- LXVFHIBXOWJTKZ-BZSNNMDCSA-N Phe-Asn-Tyr Chemical compound N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O LXVFHIBXOWJTKZ-BZSNNMDCSA-N 0.000 description 1
- JIYJYFIXQTYDNF-YDHLFZDLSA-N Phe-Asn-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC1=CC=CC=C1)N JIYJYFIXQTYDNF-YDHLFZDLSA-N 0.000 description 1
- DDYIRGBOZVKRFR-AVGNSLFASA-N Phe-Asp-Glu Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N DDYIRGBOZVKRFR-AVGNSLFASA-N 0.000 description 1
- 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 1
- WFDAEEUZPZSMOG-SRVKXCTJSA-N Phe-Cys-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CS)C(=O)N[C@@H](CO)C(O)=O WFDAEEUZPZSMOG-SRVKXCTJSA-N 0.000 description 1
- WPTYDQPGBMDUBI-QWRGUYRKSA-N Phe-Gly-Asn Chemical compound N[C@@H](Cc1ccccc1)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O WPTYDQPGBMDUBI-QWRGUYRKSA-N 0.000 description 1
- QPVFUAUFEBPIPT-CDMKHQONSA-N Phe-Gly-Thr Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O QPVFUAUFEBPIPT-CDMKHQONSA-N 0.000 description 1
- WKTSCAXSYITIJJ-PCBIJLKTSA-N Phe-Ile-Asn Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(O)=O WKTSCAXSYITIJJ-PCBIJLKTSA-N 0.000 description 1
- DVOCGBNHAUHKHJ-DKIMLUQUSA-N Phe-Ile-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O DVOCGBNHAUHKHJ-DKIMLUQUSA-N 0.000 description 1
- CWFGECHCRMGPPT-MXAVVETBSA-N Phe-Ile-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O CWFGECHCRMGPPT-MXAVVETBSA-N 0.000 description 1
- RORUIHAWOLADSH-HJWJTTGWSA-N Phe-Ile-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CC1=CC=CC=C1 RORUIHAWOLADSH-HJWJTTGWSA-N 0.000 description 1
- YCCUXNNKXDGMAM-KKUMJFAQSA-N Phe-Leu-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YCCUXNNKXDGMAM-KKUMJFAQSA-N 0.000 description 1
- RMKGXGPQIPLTFC-KKUMJFAQSA-N Phe-Lys-Asn Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O RMKGXGPQIPLTFC-KKUMJFAQSA-N 0.000 description 1
- DOXQMJCSSYZSNM-BZSNNMDCSA-N Phe-Lys-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O DOXQMJCSSYZSNM-BZSNNMDCSA-N 0.000 description 1
- JLDZQPPLTJTJLE-IHPCNDPISA-N Phe-Trp-Asp Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC2=CNC3=CC=CC=C32)C(=O)N[C@@H](CC(=O)O)C(=O)O)N JLDZQPPLTJTJLE-IHPCNDPISA-N 0.000 description 1
- JSGWNFKWZNPDAV-YDHLFZDLSA-N Phe-Val-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 JSGWNFKWZNPDAV-YDHLFZDLSA-N 0.000 description 1
- JTKGCYOOJLUETJ-ULQDDVLXSA-N Phe-Val-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 JTKGCYOOJLUETJ-ULQDDVLXSA-N 0.000 description 1
- SOACYAXADBWDDT-CYDGBPFRSA-N Pro-Ile-Arg Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O SOACYAXADBWDDT-CYDGBPFRSA-N 0.000 description 1
- FXGIMYRVJJEIIM-UWVGGRQHSA-N Pro-Leu-Gly Chemical compound OC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@@H]1CCCN1 FXGIMYRVJJEIIM-UWVGGRQHSA-N 0.000 description 1
- 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 1
- MRYUJHGPZQNOAD-IHRRRGAJSA-N Pro-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@@H]1CCCN1 MRYUJHGPZQNOAD-IHRRRGAJSA-N 0.000 description 1
- SUENWIFTSTWUKD-AVGNSLFASA-N Pro-Leu-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O SUENWIFTSTWUKD-AVGNSLFASA-N 0.000 description 1
- MHBSUKYVBZVQRW-HJWJTTGWSA-N Pro-Phe-Ile Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O MHBSUKYVBZVQRW-HJWJTTGWSA-N 0.000 description 1
- SBVPYBFMIGDIDX-SRVKXCTJSA-N Pro-Pro-Pro Chemical compound OC(=O)[C@@H]1CCCN1C(=O)[C@H]1N(C(=O)[C@H]2NCCC2)CCC1 SBVPYBFMIGDIDX-SRVKXCTJSA-N 0.000 description 1
- PRKWBYCXBBSLSK-GUBZILKMSA-N Pro-Ser-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O PRKWBYCXBBSLSK-GUBZILKMSA-N 0.000 description 1
- YIPFBJGBRCJJJD-FHWLQOOXSA-N Pro-Trp-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)[C@@H]3CCCN3 YIPFBJGBRCJJJD-FHWLQOOXSA-N 0.000 description 1
- OOZJHTXCLJUODH-QXEWZRGKSA-N Pro-Val-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H]1CCCN1 OOZJHTXCLJUODH-QXEWZRGKSA-N 0.000 description 1
- 108010025216 RVF peptide Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- FIXILCYTSAUERA-FXQIFTODSA-N Ser-Ala-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O FIXILCYTSAUERA-FXQIFTODSA-N 0.000 description 1
- OOKCGAYXSNJBGQ-ZLUOBGJFSA-N Ser-Asn-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O OOKCGAYXSNJBGQ-ZLUOBGJFSA-N 0.000 description 1
- RNMRYWZYFHHOEV-CIUDSAMLSA-N Ser-Gln-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O RNMRYWZYFHHOEV-CIUDSAMLSA-N 0.000 description 1
- LOKXAXAESFYFAX-CIUDSAMLSA-N Ser-His-Cys Chemical compound OC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CS)C(O)=O)CC1=CN=CN1 LOKXAXAESFYFAX-CIUDSAMLSA-N 0.000 description 1
- 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 1
- JEHPKECJCALLRW-CUJWVEQBSA-N Ser-His-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O JEHPKECJCALLRW-CUJWVEQBSA-N 0.000 description 1
- CJINPXGSKSZQNE-KBIXCLLPSA-N Ser-Ile-Gln Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(N)=O)C(O)=O CJINPXGSKSZQNE-KBIXCLLPSA-N 0.000 description 1
- FPCGZYMRFFIYIH-CIUDSAMLSA-N Ser-Lys-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O FPCGZYMRFFIYIH-CIUDSAMLSA-N 0.000 description 1
- XVWDJUROVRQKAE-KKUMJFAQSA-N Ser-Phe-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CO)CC1=CC=CC=C1 XVWDJUROVRQKAE-KKUMJFAQSA-N 0.000 description 1
- PCMZJFMUYWIERL-ZKWXMUAHSA-N Ser-Val-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O PCMZJFMUYWIERL-ZKWXMUAHSA-N 0.000 description 1
- 241001655322 Streptomycetales Species 0.000 description 1
- UHBPFYOQQPFKQR-JHEQGTHGSA-N Thr-Gln-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O UHBPFYOQQPFKQR-JHEQGTHGSA-N 0.000 description 1
- AQAMPXBRJJWPNI-JHEQGTHGSA-N Thr-Gly-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O AQAMPXBRJJWPNI-JHEQGTHGSA-N 0.000 description 1
- XPNSAQMEAVSQRD-FBCQKBJTSA-N Thr-Gly-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)NCC(=O)NCC(O)=O XPNSAQMEAVSQRD-FBCQKBJTSA-N 0.000 description 1
- CRZNCABIJLRFKZ-IUKAMOBKSA-N Thr-Ile-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H]([C@@H](C)O)N CRZNCABIJLRFKZ-IUKAMOBKSA-N 0.000 description 1
- 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 1
- FQPDRTDDEZXCEC-SVSWQMSJSA-N Thr-Ile-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O FQPDRTDDEZXCEC-SVSWQMSJSA-N 0.000 description 1
- IHAPJUHCZXBPHR-WZLNRYEVSA-N Thr-Ile-Tyr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)NC(=O)[C@H]([C@@H](C)O)N IHAPJUHCZXBPHR-WZLNRYEVSA-N 0.000 description 1
- XIULAFZYEKSGAJ-IXOXFDKPSA-N Thr-Leu-His Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CC1=CNC=N1 XIULAFZYEKSGAJ-IXOXFDKPSA-N 0.000 description 1
- VBMOVTMNHWPZJR-SUSMZKCASA-N Thr-Thr-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(O)=O VBMOVTMNHWPZJR-SUSMZKCASA-N 0.000 description 1
- JAWUQFCGNVEDRN-MEYUZBJRSA-N Thr-Tyr-Leu Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC(C)C)C(=O)O)N)O JAWUQFCGNVEDRN-MEYUZBJRSA-N 0.000 description 1
- YEGMNOHLZNGOCG-UBHSHLNASA-N Trp-Asn-Asn Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O YEGMNOHLZNGOCG-UBHSHLNASA-N 0.000 description 1
- HQJOVVWAPQPYDS-ZFWWWQNUSA-N Trp-Gly-Arg Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O HQJOVVWAPQPYDS-ZFWWWQNUSA-N 0.000 description 1
- WNGMGTMSUBARLB-RXVVDRJESA-N Trp-Trp-Gly Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CC=3C4=CC=CC=C4NC=3)N)C(=O)NCC(O)=O)=CNC2=C1 WNGMGTMSUBARLB-RXVVDRJESA-N 0.000 description 1
- PCXFIOFKIMNHGR-UHFFFAOYSA-N Tyr Trp Gly Ser Chemical compound C=1NC2=CC=CC=C2C=1CC(C(=O)NCC(=O)NC(CO)C(O)=O)NC(=O)C(N)CC1=CC=C(O)C=C1 PCXFIOFKIMNHGR-UHFFFAOYSA-N 0.000 description 1
- TVOGEPLDNYTAHD-CQDKDKBSSA-N Tyr-Ala-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 TVOGEPLDNYTAHD-CQDKDKBSSA-N 0.000 description 1
- AKFLVKKWVZMFOT-IHRRRGAJSA-N Tyr-Arg-Asn Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(O)=O AKFLVKKWVZMFOT-IHRRRGAJSA-N 0.000 description 1
- GAYLGYUVTDMLKC-UWJYBYFXSA-N Tyr-Asp-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 GAYLGYUVTDMLKC-UWJYBYFXSA-N 0.000 description 1
- HGEHWFGAKHSIDY-SRVKXCTJSA-N Tyr-Asp-Cys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CS)C(=O)O)N)O HGEHWFGAKHSIDY-SRVKXCTJSA-N 0.000 description 1
- CRHFOYCJGVJPLE-AVGNSLFASA-N Tyr-Gln-Asn Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)O CRHFOYCJGVJPLE-AVGNSLFASA-N 0.000 description 1
- UNUZEBFXGWVAOP-DZKIICNBSA-N Tyr-Glu-Val 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](C(C)C)C(O)=O UNUZEBFXGWVAOP-DZKIICNBSA-N 0.000 description 1
- IJUTXXAXQODRMW-KBPBESRZSA-N Tyr-Gly-His Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)NCC(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N)O IJUTXXAXQODRMW-KBPBESRZSA-N 0.000 description 1
- NMKJPMCEKQHRPD-IRXDYDNUSA-N Tyr-Gly-Tyr Chemical compound C([C@H](N)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 NMKJPMCEKQHRPD-IRXDYDNUSA-N 0.000 description 1
- QSFJHIRIHOJRKS-ULQDDVLXSA-N Tyr-Leu-Arg Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O QSFJHIRIHOJRKS-ULQDDVLXSA-N 0.000 description 1
- KSCVLGXNQXKUAR-JYJNAYRXSA-N Tyr-Leu-Glu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O KSCVLGXNQXKUAR-JYJNAYRXSA-N 0.000 description 1
- PRONOHBTMLNXCZ-BZSNNMDCSA-N Tyr-Leu-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 PRONOHBTMLNXCZ-BZSNNMDCSA-N 0.000 description 1
- WDGDKHLSDIOXQC-ACRUOGEOSA-N Tyr-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=C(O)C=C1 WDGDKHLSDIOXQC-ACRUOGEOSA-N 0.000 description 1
- 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 1
- PHKQVWWHRYUCJL-HJOGWXRNSA-N Tyr-Phe-Tyr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O PHKQVWWHRYUCJL-HJOGWXRNSA-N 0.000 description 1
- KWKJGBHDYJOVCR-SRVKXCTJSA-N Tyr-Ser-Cys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)O)N)O KWKJGBHDYJOVCR-SRVKXCTJSA-N 0.000 description 1
- IEWKKXZRJLTIOV-AVGNSLFASA-N Tyr-Ser-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(O)=O IEWKKXZRJLTIOV-AVGNSLFASA-N 0.000 description 1
- NZBSVMQZQMEUHI-WZLNRYEVSA-N Tyr-Thr-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N NZBSVMQZQMEUHI-WZLNRYEVSA-N 0.000 description 1
- NUQZCPSZHGIYTA-HKUYNNGSSA-N Tyr-Trp-Gly Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CC3=CC=C(C=C3)O)N NUQZCPSZHGIYTA-HKUYNNGSSA-N 0.000 description 1
- OJCISMMNNUNNJA-BZSNNMDCSA-N Tyr-Tyr-Asp Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC(O)=O)C(O)=O)C1=CC=C(O)C=C1 OJCISMMNNUNNJA-BZSNNMDCSA-N 0.000 description 1
- HSCJRCZFDFQWRP-UHFFFAOYSA-N Uridindiphosphoglukose Natural products OC1C(O)C(O)C(CO)OC1OP(O)(=O)OP(O)(=O)OCC1C(O)C(O)C(N2C(NC(=O)C=C2)=O)O1 HSCJRCZFDFQWRP-UHFFFAOYSA-N 0.000 description 1
- AZSHAZJLOZQYAY-FXQIFTODSA-N Val-Ala-Ser Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O AZSHAZJLOZQYAY-FXQIFTODSA-N 0.000 description 1
- QPZMOUMNTGTEFR-ZKWXMUAHSA-N Val-Asn-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](C(C)C)N QPZMOUMNTGTEFR-ZKWXMUAHSA-N 0.000 description 1
- DBOXBUDEAJVKRE-LSJOCFKGSA-N Val-Asn-Val Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](C(C)C)C(=O)O)N DBOXBUDEAJVKRE-LSJOCFKGSA-N 0.000 description 1
- OQWNEUXPKHIEJO-NRPADANISA-N Val-Glu-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CO)C(=O)O)N OQWNEUXPKHIEJO-NRPADANISA-N 0.000 description 1
- SYOMXKPPFZRELL-ONGXEEELSA-N Val-Gly-Lys Chemical compound CC(C)[C@@H](C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)O)N SYOMXKPPFZRELL-ONGXEEELSA-N 0.000 description 1
- 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 1
- AIWLHFZYOUUJGB-UFYCRDLUSA-N Val-Phe-Tyr Chemical compound C([C@H](NC(=O)[C@@H](N)C(C)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=CC=C1 AIWLHFZYOUUJGB-UFYCRDLUSA-N 0.000 description 1
- UVHFONIHVHLDDQ-IFFSRLJSSA-N Val-Thr-Glu Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](C(C)C)N)O UVHFONIHVHLDDQ-IFFSRLJSSA-N 0.000 description 1
- DFQZDQPLWBSFEJ-LSJOCFKGSA-N Val-Val-Asn Chemical compound CC(C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(=O)N)C(=O)O)N DFQZDQPLWBSFEJ-LSJOCFKGSA-N 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 108010045023 alanyl-prolyl-tyrosine Proteins 0.000 description 1
- 108010070944 alanylhistidine Proteins 0.000 description 1
- 230000002785 anti-thrombosis Effects 0.000 description 1
- 230000010100 anticoagulation Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 108010040443 aspartyl-aspartic acid Proteins 0.000 description 1
- 108010069205 aspartyl-phenylalanine Proteins 0.000 description 1
- 108010068265 aspartyltyrosine Proteins 0.000 description 1
- 239000000022 bacteriostatic agent Substances 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 238000007036 catalytic synthesis reaction Methods 0.000 description 1
- 239000003610 charcoal Substances 0.000 description 1
- 238000005352 clarification Methods 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- FSXRLASFHBWESK-UHFFFAOYSA-N dipeptide phenylalanyl-tyrosine Natural products C=1C=C(O)C=CC=1CC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FSXRLASFHBWESK-UHFFFAOYSA-N 0.000 description 1
- 108010054812 diprotin A Proteins 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 238000001641 gel filtration chromatography Methods 0.000 description 1
- 229940097043 glucuronic acid Drugs 0.000 description 1
- 108010079547 glutamylmethionine Proteins 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 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 1
- XKUKSGPZAADMRA-UHFFFAOYSA-N glycyl-glycyl-glycine Natural products NCC(=O)NCC(=O)NCC(O)=O XKUKSGPZAADMRA-UHFFFAOYSA-N 0.000 description 1
- 108010051307 glycyl-glycyl-proline Proteins 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000004191 hydrophobic interaction chromatography Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000007603 infrared drying Methods 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 108010076756 leucyl-alanyl-phenylalanine Proteins 0.000 description 1
- 108010034529 leucyl-lysine Proteins 0.000 description 1
- 108010044056 leucyl-phenylalanine Proteins 0.000 description 1
- 108010012058 leucyltyrosine Proteins 0.000 description 1
- 238000004246 ligand exchange chromatography Methods 0.000 description 1
- 229940127215 low-molecular weight heparin Drugs 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 108010054155 lysyllysine Proteins 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 108010016686 methionyl-alanyl-serine Proteins 0.000 description 1
- 108010056582 methionylglutamic acid Proteins 0.000 description 1
- 108010085203 methionylmethionine Proteins 0.000 description 1
- 238000001471 micro-filtration Methods 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 239000002304 perfume Substances 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 108010084572 phenylalanyl-valine Proteins 0.000 description 1
- 108010073025 phenylalanylphenylalanine Proteins 0.000 description 1
- 108010083476 phenylalanyltryptophan Proteins 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108010053725 prolylvaline Proteins 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000001223 reverse osmosis Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 108010069117 seryl-lysyl-aspartic acid Proteins 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000001694 spray drying Methods 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 230000019635 sulfation Effects 0.000 description 1
- 238000005670 sulfation reaction Methods 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 108010051110 tyrosyl-lysine Proteins 0.000 description 1
- 108010003137 tyrosyltyrosine Proteins 0.000 description 1
- 241001446247 uncultured actinomycete Species 0.000 description 1
- 229940118696 vibrio cholerae Drugs 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
Images
Classifications
-
- 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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
-
- 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
- C12N9/10—Transferases (2.)
- C12N9/1048—Glycosyltransferases (2.4)
- C12N9/1051—Hexosyltransferases (2.4.1)
-
- 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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/18—Preparation of compounds containing saccharide radicals produced by the action of a glycosyl transferase, e.g. alpha-, beta- or gamma-cyclodextrins
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
本发明公开了一种岩藻糖基化软骨素及其制备方法和应用。本发明利用酶的底物宽泛性,筛选不同来源的岩藻糖基转移酶进行底物适应性研究,构建多酶级联催化反应体系,开发了高效的酶法合成岩藻糖基化软骨素的方法,采用本发明的方法可以在软骨素糖链的不同位点实现不同程度的岩藻糖基化修饰,为岩藻糖基硫酸软骨素的工业化生产奠定了基础,推动糖类抗凝药物的开发。
Description
技术领域
本发明涉及酶工程技术及合成生物学领域,特别涉及一种岩藻糖基化软骨素及其制备方法和应用。
背景技术
随着人类生活水平的普遍提高和人口老龄化进程的加快,血栓性疾病日益增多,已成为一个严重影响人类生命健康的问题。肝素和低分子肝素(LMWHs)作为抗凝剂被广泛应用于血栓性疾病的治疗。肝素和低分子肝素在治疗期间有严重出血的风险,并且不能口服。因此,研究人员们一直在继续寻找更有效和更安全的抗凝药物。岩藻糖基化硫酸软骨素(FuCS)是一种从海参中分离出来的结构独特的活性多糖,具有显著的抗凝和抗血栓活性,可以作为一种潜在的候选药物,其合成制备以及生物学功能研究成为了国内外科学家关注的热点。然而,由于均质寡糖的缺乏,阻碍了进一步的构效关系研究。
目前,FuCS的获得主要有海参提取和化学法合成两种方法。提取法受海参品质和来源的限制,无法获得结构明确的FuCS糖链,限制了构效关系和生物研究(如ACS ChemBiol 2020,15(8),2232-2246.或CarbohydrPolym 2017,165,7-12.或CarbohydrPolym2021,262, 117969.);化学合成FuCS近年来得到了长足发展,但受糖链复杂性影响,依然存在步骤繁琐、收率低等问题,无法做到经济高效合成(如Angew Chem Int Ed Engl 2018,57(39), 12880-12885.或J Org Chem 2020,85,24,15908–15919.)。
FuCS糖链结构是由硫酸化修饰的二糖重复单元与GlcA O-3位分支修饰岩藻糖残基构成,其二糖重复单元主要由糖醛酸(葡萄糖醛酸GlcA)和N-乙酰氨基半乳糖(GalNAc)通过β糖苷键连接构成。不同物种来源的FuCS在硫酸化位点及岩藻糖基化分支的数目上存在差异,目前现有技术中尚未有酶法合成FuCS寡糖片段的相关报道,鉴于此,特提出本发明。
发明内容
本发明克服了现有技术中存在的不足,提供了一种岩藻糖基化软骨素及其制备方法和应用。
本发明的第一方面提供了一种岩藻糖基化软骨素的制备方法,所述的方法包括如下步骤:
将L-岩藻糖基转移酶与底物GDP-L-岩藻糖和软骨素寡糖混合,酶促反应,得到岩藻糖基化软骨素。
进一步地,所述软骨素寡糖中可岩藻糖基化的GlcA与GDP-L-岩藻糖(鸟苷二磷酸岩藻糖)的摩尔比为0.5-2:1-4,优选地,所述摩尔比为0.5-1.5:1-3.5,更优选地,所述摩尔比为 1:1-3.5(如1:1、1:1.5、1:2、1:2.5、1:3、1:3.5)。
在本发明的一个具体实施方式中,所述软骨素寡糖中可岩藻糖基化的GlcA与GDP-L- 岩藻糖(鸟苷二磷酸岩藻糖)的摩尔比为1:2。
进一步地,所述反应的pH为6.5-8.5,优选地,所述反应的pH为7-8,更优选地,所述反应的pH为7.5。
进一步地,所述反应的温度为30-45℃,优选的,所述反应的温度为35-40℃,更优选的,所述反应的温度为37℃。
进一步地,所述反应的时间为10-30h,优选的,所述反应的时间为10-25h,更优选的,所述反应的时间为12-24h。
进一步地,所述L-岩藻糖基转移酶的浓度为0.1-2mg/mL,优选的,所述L-岩藻糖基转移酶的浓度为0.5-1.5mg/mL,更优选的,所述L-岩藻糖基转移酶的浓度为1mg/mL。
进一步地,所述的软骨素寡糖具有如下结构:
进一步地,所述n1为选自1-10的整数(如1、2、3、4、5、6、7、8、9、10)。
优选地,所述的n1为选自1-5的整数,更优选地,所述的n1为选自1-3的整数。
在本发明的一个具体实施方式中,所述的软骨素寡糖选自如下结构:
进一步地,所述L-岩藻糖基转移酶的核苷酸序列如SEQ ID NO:1或SEQ ID NO:2所示,氨基酸序列如SEQ ID NO:3或SEQ ID NO:4所示;
进一步地,所述的L-岩藻糖基转移酶Bfα1,3FT的核苷酸序列如SEQ ID NO:1所示,氨基酸序列如SEQ ID NO:3所示;
进一步地,所述的L-岩藻糖基转移酶Hpα1,3FT S45F V30G的核苷酸序列如SEQ IDNO: 2所示,氨基酸序列如SEQ ID NO:4所示。
进一步地,所述的L-岩藻糖基转移酶选自α-1,2-岩藻糖基转移酶或α-1,3-岩藻糖基转移酶中的一种。
优选地,所述的L-岩藻糖基转移酶为α-1,3-岩藻糖基转移酶。
优选地,所述的L-岩藻糖基转移酶选自:SEQ ID NO:3-4中任一项所表示的多肽、SEQ ID NO3-4中任一项所表示的多肽的功能性变体、SEQ ID NO:3-4中的任一项所表示的多肽的功能性片段以及SEQ ID NO:3-4中任一项所表示的多肽的功能性片段的功能性变体。
进一步地,所述L-岩藻糖基转移酶的来源为:脆弱拟杆菌(Bacteroidesfragilis)、幽门螺杆菌(Helicobacter pylori)、霍乱弧菌(Vibriocholera)、胆型螺杆菌(H.bilis)、朱奈德螺杆菌 (H.cinaede)、卵形拟杆菌(Bacteroides ovatus)和普通拟杆菌(Bacteroides vulgatus)中的一种,优选地,所述L-岩藻糖基转移酶的来源为脆弱拟杆菌(Bacteroides fragilis)和/或幽门螺杆菌 (Helicobacter pylori)。
在本发明的一个具体实施方式中,所述的L-岩藻糖基转移酶为Bfα1,3FT(来源于脆弱拟杆菌,Bacteroides fragilis)。
在本发明的一个具体实施方式中,所述的L-岩藻糖基转移酶为Hpα1,3FT(来源于幽门螺杆菌,Helicobacter pylori)。
在本发明的一个具体实施方式中,所述的L-岩藻糖基转移酶为Hpα1,3FT S45FV30G突变体,其催化活性有所提高。
Bfα1,3FT可识别寡糖内所有的D-GalNAc1-4-β-D-GlcA二糖单元并在GlcA O-3位进行岩藻糖基化修饰,而Hpα1,3FT仅可识别非还原端为D-GalNAc1-4-β-D-GlcA结构的寡糖并且仅对其非还原端的GlcA O-3位进行单位点的岩藻糖基化修饰。Bfα1,3FT催化识别非还原端为 D-GlcA1-3-β-D-GalNAc结构的寡糖时,非还原端大于等于两个糖未经岩藻糖基化修饰,可由软骨素合成酶催化其继续延伸,并根据需要在特定位置进一步进行岩藻糖基化修饰。
在本发明的一个具体实施方式中,所述的Hpα1,3FT S45F V30G突变体可识别的软骨素寡糖结构如式(Ⅰ)所示:
在本发明的一个具体实施方式中,所述的Hpα1,3FT S45F V30G突变体可识别的软骨素寡糖的结构分别如式(IV)、(VI)、(Ⅷ)所示:
在本发明的一个具体实施方式中,所述的Hpα1,3FT S45F V30G突变体催化合成的单一位点岩藻糖基化修饰的软骨素寡糖的结构分别如式(Ⅸ)、(X)、(XI)所示:
在本发明的一个具体实施方式中,所述Bfα1,3FT可识别的软骨素寡糖结构分别如式(Ⅰ) 和(Ⅱ)所示:
在本发明的一个具体实施方式中,所述Bfα1,3FT可识别的软骨素寡糖结构分别如式(IV)、 (V)、(VI)、(VII)、(VIII)所示:
在本发明的一个具体实施方式中,所述Bfα1,3FT催化合成的岩藻糖基化软骨素寡糖结构分别如式(Ⅸ)、(Ⅻ)、(XIII)、(XIV)、(XV)所示:
进一步地,所述的制备方法还包括分离、纯化的步骤。
进一步地,当岩藻糖基化软骨素寡糖的非还原端大于等于两个糖未岩藻糖基化修饰,软骨素合成酶PmCS可催化主链进一步延伸,并根据需要在特定位置由Bfα1,3FT或 Hpα1,3FTS45F V30G催化进一步岩藻糖基化修饰,合成结构确定、岩藻糖基化程度确定的软骨素寡糖。
进一步地,所述L-岩藻糖基转移酶Hpα1,3FT S45F V30G催化非还原端结构为 D-GalNAc1-4-β-D-GlcA的GlcA O-3位进行单位点的岩藻糖基化修饰。
进一步地,所述L-岩藻糖基转移酶Bfα1,3FT催化结构为D-GalNAc1-4-β-D-GlcA的所有 GlcA O-3位进行岩藻糖基化修饰。进一步地,所述的延伸岩藻糖基化软骨素主链的步骤包括将所述主链与软骨素合成酶PmCS、糖核苷酸混合,反应。
进一步地,所述的糖核苷酸选自:UDP-GalNAc、UDP-GlcA。
在本发明的一个实施方式中,所述岩藻糖基化软骨素的制备方法包括如下步骤:首先使用软骨素合成酶延伸岩藻糖基化软骨素的主链,然后使用L-岩藻糖基转移酶Hpα1,3FT S45F V30G或Bfα1,3FT催化特定位点的岩藻糖基化修饰。
在本发明的一个实施方式中,所述岩藻糖基化软骨素的制备方法包括如下步骤:首先使用软骨素合成酶延伸岩藻糖基化软骨素的主链,然后在非还原端结构为 D-GalNAc1-4-β-D-GlcA的GlcA O-3位使用L-岩藻糖基转移酶Hpα1,3FT S45F V30G催化其进一步进行单位点的岩藻糖基化修饰。
在本发明的一个实施方式中,所述岩藻糖基化软骨素的制备方法包括如下步骤:首先使用软骨素合成酶延伸岩藻糖基化软骨素的主链,然后在结构为D-GalNAc1-4-β-D-GlcA的所有GlcA O-3位使用L-岩藻糖基转移酶Bfα1,3FT催化其进一步岩藻糖基化修饰。
在本发明的一个具体实施方式中,可由软骨素合成酶PmCS进一步延伸的岩藻糖基化软骨素主链的结构分别如(Ⅻ)、(XIV)所示:
在本发明的一个具体实施方式中,所述岩藻糖基化软骨素主链延伸后的结构分别如 (XVI)、(XVII)、(XVIII)、(XIX)所示:
在本发明的一个具体实施方式中,使用软骨素合成酶延伸岩藻糖基化软骨素主链的方法包括如下步骤:使用软骨素合成酶PmCS催化岩藻糖基化软骨素(Ⅻ)进一步进行岩藻糖基化修饰,合成岩藻糖基化软骨素(XVI)。
在本发明的一个具体实施方式中,使用软骨素合成酶延伸岩藻糖基化软骨素主链的方法包括如下步骤:使用软骨素合成酶PmCS催化岩藻糖基化软骨素(XVI)进一步进行岩藻糖基化修饰,合成岩藻糖基化软骨素(XVIII)。
在本发明的一个具体实施方式中,使用软骨素合成酶延伸岩藻糖基化软骨素主链的方法包括如下步骤:使用软骨素合成酶PmCS催化岩藻糖基化软骨素(XVIII)进一步进行岩藻糖基化修饰,合成岩藻糖基化软骨素(XIX)。
在本发明的一个具体实施方式中,使用软骨素合成酶延伸岩藻糖基化软骨素主链的方法包括如下步骤:使用软骨素合成酶PmCS催化岩藻糖基化软骨素(XIV)进一步进行岩藻糖基化修饰,合成岩藻糖基化软骨素(XVII)。
进一步地,所述的L-岩藻糖基转移酶Hpα1,3FTS45F V30G可识别延伸后的岩藻糖基化软骨素,催化其进一步进行岩藻糖基化修饰。
在本发明的一个具体实施方式中,所述的岩藻糖基化软骨素的制备方法包括如下步骤:使用L-岩藻糖基转移酶Hpα1,3FTS45F V30G催化岩藻糖基化软骨素(XⅥ)进一步进行岩藻糖基化修饰,合成岩藻糖基化软骨素(XⅢ)。
在本发明的一个具体实施方式中,所述的岩藻糖基化软骨素的制备方法包括如下步骤:使用L-岩藻糖基转移酶Hpα1,3FTS45F V30G催化岩藻糖基化软骨素(XVII)进一步进行岩藻糖基化修饰,合成岩藻糖基化软骨素(XV)。
在本发明的一个具体实施方式中,所述的岩藻糖基化软骨素的制备方法包括如下步骤:使用L-岩藻糖基转移酶Hpα1,3FTS45F V30G催化岩藻糖基化软骨素(XⅨ)进一步进行岩藻糖基化修饰,合成岩藻糖基化软骨素(XX)。
进一步地,所述的L-岩藻糖基转移酶Bfα1,3FT可识别延伸后的岩藻糖基化软骨素,催化其进一步进行岩藻糖基化修饰。
在本发明的一个具体实施方式中,所述的岩藻糖基化软骨素的制备方法包括如下步骤:使用L-岩藻糖基转移酶Bfα1,3FT催化岩藻糖基化软骨素(XⅧ)进一步进行岩藻糖基化修饰,合成岩藻糖基化软骨素(XIV)。
在本发明的一个具体实施方式中,所述的岩藻糖基化软骨素的制备方法包括如下步骤:使用L-岩藻糖基转移酶Bfα1,3FT催化岩藻糖基化软骨素(XIX)进一步进行岩藻糖基化修饰,合成岩藻糖基化软骨素(XV)。
进一步地,所述的软骨素寡糖的制备方法包括如下步骤:将软骨素合成酶与供体底物和受体底物混合,反应,得到软骨素寡糖。
进一步地,所述的供体底物(即糖核苷酸)选自:UDP-GalNAc、UDP-GlcA。
进一步地,所述的受体底物选自:GalNAc。
本发明所述的UDP-GalNAc:中文全称为UDP-N-乙酰半乳糖胺,其作用是为软骨素寡糖或软骨素多糖的合成提供活化的乙酰半乳糖胺供体(供体底物)。
本发明所述的UDP-GlcA:中文全称为UDP-葡萄糖醛酸,其作用是为软骨素寡糖或软骨素多糖的合成提供活化的葡萄糖醛酸供体(供体底物)。
进一步地,所述的单糖GalNAc与糖核苷酸的摩尔比为0.5-2:1-5,优选地,所述的软骨素寡糖与糖核苷酸的摩尔比为0.5-1.5:1-5(如0.5:1-5、0.6:1-5、0.7:1-5、0.8:1-5、0.9:1-5、1:1-5、 1.1:1-5、1.2:1-5、1.3:1-5、1.4:1-5、1.5:1-5),更优选地,所述的软骨素寡糖与糖核苷酸的摩尔比为1:1-5(如1:1、1:1.5、1:2、1:2.5、1:3、1:3.5、1:4、1:5)。
在本发明的一个具体实施方式中,所述的软骨素寡糖受体与糖核苷酸的摩尔比为1:1.5-1:2。
进一步地,所述的软骨素合成酶来源于:多杀巴斯德杆菌(Pasteurellamultocida)、大肠杆菌(Escherichia coli)、尿放线杆菌(ActinobacillusUreae)。
进一步地,所述方法还包括分离的步骤。
进一步地,所述的分离选自:离子交换树脂、分子筛、澄清、超滤、纳米过滤、反渗透、微滤、活性木炭或碳处理、切向流高效过滤、切向流超滤、亲和色谱、离子交换色谱、疏水相互作用色谱和/或凝胶过滤和配体交换色谱中的一种。
进一步地,所述的分子筛选自:聚丙烯酰胺分子筛(如Bio-Gel P2到P300)、琼脂糖分子筛、葡聚糖分子筛。
进一步地,所述的离子交换树脂为阴离子交换树脂,所述的阴离子交换树脂为强碱性阴离子交换树脂或弱碱性阴离子交换树脂,优选地,所述的阴离子交换树脂为强碱性阴离子交换树脂。
进一步地,所述的强碱性阴离子交换树脂为凝胶型强碱性阴离子交换树脂。
在本发明的一个具体实施方式中,所述的分离为阴离子交换树脂。
在本发明的一个具体实施方式中,所述的分离为强碱性阴离子交换树脂。
在本发明的一个具体实施方式中,所述的分离为分子筛。
在本发明的一个具体实施方式中,所述的分离为聚丙烯酰胺分子筛。
在本发明的一个具体实施方式中,所述的分离为Bio-Gel P2分子筛。
进一步地,所述方法还包括纯化的步骤。
进一步地,所述的纯化包括使用碱、盐、缓冲溶液或酸的适当稀释的溶液洗涤软骨素寡糖的步骤。
进一步地,所述的纯化还包括使用活性木炭或碳,使用木炭,纳米过滤,超滤或离子交换,使用醇,使用含水醇混合物进行结晶,蒸发,浓缩、沉淀,干燥的步骤。
进一步地,所述的浓缩选自:常压浓缩、减压浓缩、薄膜浓缩、多效浓缩,所述干燥选自:冷冻干燥、常压干燥、减压干燥、沸腾干燥、喷雾干燥、红外线干燥、微波干燥。
在本发明的一个具体实施方式中,所述的干燥为冷冻干燥。
进一步地,所述的岩藻糖基化软骨素,其具有如下结构:
在本发明的一个具体实施方式中,所述的岩藻糖基化软骨素具有如下结构:
本发明的第二方面提供了一种岩藻糖基化软骨素,其具有如下结构:
在本发明的一个具体实施方式中,所述的岩藻糖基化软骨素具有如下结构:
本发明的第三方面提供了两种岩藻糖基转移酶,所述岩藻糖基转移酶的核苷酸序列如SEQ ID NO:1或SEQ ID NO:2所示,氨基酸序列如SEQ ID NO:3或SEQ ID NO: 4所示。
本发明的第四方面提供了一种重组载体,所述重组载体是由在质粒载体中***如第三方面所述的如SEQ ID NO:1或SEQ ID NO:2所示的岩藻糖基转移酶的核苷酸序列构建得到的。
进一步地,所述的载体为病毒载体或非病毒载体。在本发明的一个实施方式中,所述的载体为质粒载体。
在本发明的一个实施方式中,所述的载体为PMAL-C2X。
在本发明的一个实施方式中,所述的载体为pET-41a。
本发明的第五方面提供了一种重组细胞。
进一步地,所述的重组细胞选自:***、革兰氏阴性细菌、放线菌、真菌;
优选地,所述的***选自:枯草芽孢杆菌;
优选地,所述的革兰氏阴性细菌选自:大肠杆菌;所述的放线菌为链霉菌;
优选地,所述的真菌为酵母类、曲酶菌。
本发明的第六方面提供了一种包含如第二方面所述的岩藻糖基化软骨素的药物组合物。
进一步地,所述的药物组合物还包括药学上可接受的辅料。
进一步的,所述的药学上可接受的辅料选自载体、赋形剂、稀释剂、润滑剂、润湿剂、乳化剂、防腐剂、抗氧化剂、缓冲剂、抑菌剂、使制剂与接受者的血液等渗的溶质、悬浮剂、助悬剂、增溶剂、增稠剂、稳定剂、甜味剂以及香料中的一种或两种以上的组合。
本发明的第七方面提供了一种如第二方面所述的岩藻糖基化软骨素或如第六方面所述的药物组合物在制备抗凝血药物或岩藻糖基化硫酸软骨素中的用途。
本发明的第八方面提供了一种如第三方面所述的岩藻糖基转移酶在合成岩藻糖基化软骨素中的应用。
本发明的第九方面提供了如第二方面所述的一种岩藻糖基化软骨素在制备岩藻糖基化硫酸软骨素中的应用。
本发明所述的生物酶(岩藻糖基转移酶和软骨素合成酶)来自原核生物,具有蛋白表达量高、底物适应性宽、催化效率高等优点。酶法合成过程具有高效性及高区域选择性,操作简单且安全,为岩藻糖基硫酸软骨素的工业化生产奠定了基础,解决酶法合成岩藻糖基化软骨素的瓶颈问题,提供体外多酶级联制备岩藻糖基化软骨素的方法,本发明利用酶的底物宽泛性,筛选不同来源的岩藻糖基转移酶进行底物适应性研究,开发了高效的酶法合成岩藻糖基化软骨素的方法,采用本发明的方法可对底物软骨素寡糖进行不同程度的岩藻糖基化修饰,寡糖非还原端为岩藻糖基硫酸软骨素的工业化生产奠定了基础,推动抗凝药物的开发。
附图说明
图1为岩藻糖基化软骨素的制备示意图。
具体实施方式
为了能够更清楚地理解本发明的技术内容,特举以下实施例详细说明,其目的仅在于更好理解本发明的内容而非限制本发明的保护范围。
本发明中所述的式(Ⅸ)~式(XX)结构分别如下所示:
实施例1Hpα1,3FT S45F V30G催化合成单一位点岩藻糖基化修饰的软骨素寡糖
(1)将鸟苷二磷酸岩藻糖(GDP-Fuc)、软骨素三糖(IV)、MgCl2、Tris-HCl配制成水溶液,反应体系中GDP-Fuc、MgCl2的终浓度为20mM,软骨素三糖(IV)的终浓度为10mM,Tris-HCl的终浓度为50mM,反应体系pH为7.5,然后添加L-岩藻糖基转移酶,反应12h,检测反应完成后,加入冰乙醇终止反应;将反应液离心10-20min,将上清通过Bio-Gel P2 分子筛脱盐后,利用强碱性阴离子交换树脂介质分离纯化,再次利用Bio-Gel P2分子筛脱盐后,得到产物IX,转化率达到90%,其1H NMR、13C NMR及MS数据如下,
1H NMR(600MHz,D2O)δ5.40(d,J=4.0Hz,2H,H-1Fuc),5.22(d,J=3.8Hz,1H, H-1GalNAc-A),4.69(d,J=8.5Hz,1H,H-1GalNAc-A),4.57(d,J=8.0Hz,1H),4.52(d,J=7.9Hz,1H),4.46(dd,J=8.4,2.5Hz,2H),4.29(dd,J=11.1,3.8Hz,1H),4.22(d,J=3.1Hz,1H),4.16-4.11(m,2H),4.02-3.89(m,11H),3.83(d,J=3.3Hz,2H),3.81-3.64(m,19H),3.61-3.56(m, 4H),2.05(dd,J=26.0,2.3Hz,12H,COOCH3,COOCH3),1.25(d,J=6.6Hz,6H,CH3 ofFucose).
13C NMR(151MHz,D2O)δ175.42,175.40,175.06,174.92,174.62,104.01,103.85,100.00, 98.31,95.17,91.19,80.77,77.73,76.73,76.67,76.62,75.22,75.18,74.95,74.91,74.22,74.19, 74.16,72.06,71.54,70.25,69.25,68.42,68.13,67.71,67.53,66.66,61.50,61.26,61.05,52.24, 48.83,22.47,22.22,22.01,15.37.
ESI-MS(negative mode)calcd for C28H45N2O21 -[M-H]-m/z745.2520,found745.2527.;
(2)将鸟苷二磷酸岩藻糖(GDP-Fuc)、软骨素五糖(VI)、MgCl2、Tris-HCl配制成水溶液,反应体系中GDP-Fuc、MgCl2的终浓度为20mM,软骨素五糖(VI)的终浓度为10mM,Tris-HCl的终浓度为50mM,反应体系pH为7.5,然后添加L-岩藻糖基转移酶,反应12h,检测反应完成后,加入冰乙醇终止反应;将反应液离心10-20min,将上清通过Bio-Gel P2 分子筛脱盐后,利用强碱性阴离子交换树脂介质分离纯化,再次利用Bio-Gel P2分子筛脱盐后,得到产物X,转化率达到86%,其1H NMR、13C NMR及MS数据如下,
1H NMR(600MHz,D2O)δ5.39(d,J=3.9Hz,2H,H-1Fuc),5.22(d,J=3.7Hz,1H, H-1GalNAc-A),4.68(d,J=8.5Hz,1H,H-1GalNAc-A),4.58-4.43(m,8H,H-1GlcA-A,H-1GalNAc-B, H-1GlcA -B,H-1GalNAc-C),4.29(dd,J=11.1,3.7Hz,1H),4.20(d,J=3.1Hz,1H),4.15-4.11(m,4H),4.03-3.89(m,12H),3.83-3.68(m,31H),3.67-3.54(m,9H),3.40-3.36(m,2H),2.09-2.00(m,18H, COOCH3,COOCH3,COOCH3),1.24(d,J=6.5Hz,6H,CH3 of Fucose).
13C NMR(151MHz,D2O)δ175.40,175.06,174.98,174.93,174.67,174.58,104.28,104.10, 104.03,100.84,99.95,98.28,95.18,91.18,80.70,80.34,79.71,77.32,76.59,76.52,76.38,75.13, 74.95,74.91,74.20,73.79,73.75,72.49,72.42,72.07,71.57,70.30,69.24,68.51,68.11,67.83, 67.60,67.53,66.62,61.50,61.26,61.08,61.04,52.31,52.24,50.88,48.91,22.48,22.45,22.22, 21.99,15.35.
ESI-MS(negative mode)calcd for C42H66N3O32 -[M-H]-m/z1124.3635,found1124.3399; calcd for C42H65N3O32 2-[M-2H]2-m/z561.6781,found 561.6675.;
(3)将鸟苷二磷酸岩藻糖(GDP-Fuc)、软骨素七糖(VIII)、MgCl2、Tris-HCl配制成水溶液,反应体系中GDP-Fuc、MgCl2的终浓度为20mM,软骨素七糖(VIII)的终浓度为 10mM,Tris-HCl的终浓度为50mM,反应体系pH为7.5,然后添加L-岩藻糖基转移酶,反应12h,检测反应完成后,加入冰乙醇终止反应;将反应液离心10-20min,将上清通过 Bio-Gel P2分子筛脱盐后,利用强碱性阴离子交换树脂介质分离纯化,再次利用Bio-Gel P2 分子筛脱盐后,得到产物XI,转化率达到90%,其1H NMR、13C NMR及MS数据如下,
1H NMR(600MHz,D2O)δ5.39(d,J=4.0Hz,2H,H-1Fuc),5.22(d,J=3.7Hz,1H, H-1GalNAc-A),4.68(d,J=8.5Hz,1H,H-1GalNAc-A),4.58-4.43(m,12H,H-1GlcA-A,H-1GalNAc-B, H-1GlcA-B,H-1GalNAc-C,H-1GlcA-C,H-1GalNAc-D),4.29(dd,J=11.1,3.8Hz,1H),4.20(d,J=3.1Hz,1H),4.14-4.12(m,6H),4.02-3.88(m,13H),3.84-3.67(m,43H),3.67-3.63(m,2H),3.62-3.54(m, 8H),3.41-3.33(m,4H),2.08-2.00(m,24H,COOCH3,COOCH3,COOCH3,COOCH3),1.24(d,J= 6.5Hz,6H,CH3 of Fucose).
13C NMR(151MHz,D2O)δ175.38,175.07,175.00,174.98,174.93,174.67,174.50,104.31, 104.29,104.11,104.04,100.87,100.81,99.94,98.28,95.18,91.18,80.72,80.32,79.75,79.66, 77.30,76.57,76.48,76.32,76.26,75.13,74.95,74.91,74.20,73.79,73.76,73.68,72.48,72.44, 72.07,72.04,71.58,70.31,69.24,68.52,68.37,68.11,67.84,67.71,67.64,67.61,67.53,66.61, 61.49,61.26,61.07,52.32,52.25,50.97,50.89,48.92,22.45,22.21,22.13,21.99,15.35.
ESI-MS(negative mode)calcd for C56H86N4O43 2-[M-2H]2-m/z751.2338,found751.2213.。
实施例2Bfα1,3FT催化合成岩藻糖基化修饰的软骨素寡糖
(1)将鸟苷二磷酸岩藻糖(GDP-Fuc)、软骨素四糖(V)、MgCl2、Tris-HCl配制成水溶液,反应体系中GDP-Fuc、MgCl2的终浓度为20mM,软骨素四糖(V)的终浓度为10mM, Tris-HCl的终浓度为50mM,反应体系pH为7.5,然后添加L-岩藻糖基转移酶Bfα1,3FT,反应12h,检测反应完成后,加入冰乙醇终止反应;将反应液离心10-20min,将上清通过Bio-Gel P2分子筛脱盐后,利用强碱性阴离子交换树脂介质分离纯化,再次利用Bio-Gel P2 分子筛脱盐后,得到产物XII,其1H NMR、13C NMR及MS数据如下,
1H NMR(600MHz,D2O)δ5.36(d,J=3.9Hz,2H,H-1Fuc),5.17(d,J=3.7Hz,1H, H-1GalNAc-A),4.63(d,J=8.5Hz,1H,H-1GalNAc-A),4.53-4.41(m,6H,H-1GlcA-A,H-1GalNAc-B, H-1GlcA -B),4.24(dd,J=11.2,3.8Hz,1H),4.17(d,J=3.0Hz,1H),4.11-4.06(m,4H,),4.03-3.98(m,2H),3.97-3.86(m,6H),3.79(d,J=3.3Hz,2H),3.76-3.62(m,22H),3.55-3.51(m,4H),3.47-3.41(m,4H),3.28(t,J=8.4Hz,2H),1.99(dd,J=13.5,2.3Hz,12H,COOCH3,COOCH3),1.21(d,J=6.6Hz,6H,CH3 of Fucose).
13C NMR(151MHz,D2O)δ175.85,175.20,175.18,174.83,174.78,174.49,103.96,103.81, 99.69,98.18,95.04,91.06,80.66,80.52,80.50,77.61,76.48,76.46,76.40,76.08,75.14,74.84, 74.80,74.58,74.11,74.07,72.62,71.93,71.71,70.14,69.06,68.31,67.97,67.59,67.47,66.55, 61.47,61.15,60.93,59.17,52.08,50.78,48.70,22.39,22.09,21.87,15.24.
ESI-MS(negative mode)calcd for C34H53N2O27 -[M-H]-m/z921.2841,found921.2821.;
(2)将鸟苷二磷酸岩藻糖(GDP-Fuc)、软骨素五糖(VI)、MgCl2、Tris-HCl配制成水溶液,反应体系中GDP-Fuc的终浓度为40mM,MgCl2的终浓度为20mM,软骨素五糖(VI) 的终浓度为10mM,Tris-HCl的终浓度为50mM,反应体系pH为7.5,然后添加L-岩藻糖基转移酶Bfα1,3FT,反应24h,检测反应完成后,加入冰乙醇终止反应;将反应液离心10-20 min,将上清通过Bio-Gel P2分子筛脱盐后,利用强碱性阴离子交换树脂介质分离纯化,再次利用Bio-Gel P2分子筛脱盐后,得到产物XIII,其1H NMR、13C NMR及MS数据如下,
1H NMR(600MHz,D2O)δ5.36(t,J=3.6Hz,4H,,H-1Fuc-A,H-1Fuc-B),5.18(d,J=3.7Hz, 1H,H-1GalNAc-A),4.65(d,J=8.5Hz,1H,H-1GalNAc-A),4.53-4.38(m,8H,H-1GlcA-A,H-1GalNAc-B, H-1GlcA-B,H-1GalNAc-C),4.25(dd,J=11.0,3.9Hz,1H),4.18(d,J=3.1Hz,1H),4.12-4.04(m,4H), 4.03-3.97(m,2H),3.97-3.85(m,15H),3.8-3.78(m,4H),3.77-3.65(m,30H),3.62(dd,J=10.7, 3.2Hz,2H),3.57-3.50(m,9H),2.05-1.98(m,18H,COOCH3,COOCH3,COOCH3),1.21(dd,J= 6.6,1.9Hz,12H,CH3 of Fucose-A,CH3 of Fucose-B).
13C NMR(151MHz,D2O)δ175.33,175.19,175.17,174.95,174.75,174.50,103.96,103.86, 103.81,99.83,99.65,98.19,98.15,95.07,91.07,81.05,80.66,77.60,76.47,76.42,75.02,74.85, 74.83,74.51,74.12,71.96,71.93,71.44,70.16,69.13,69.08,68.32,68.00,67.61,67.42,67.39, 66.56,66.52,61.49,61.41,61.17,60.94,52.13,50.66,48.72,22.39,22.35,22.11,21.88,15.25.
ESI-MS(negative mode)calcd for C48H76N3O36 -[M-H]-m/z 1270.4214,found1270.3972; calcd for C48H75N3O36 2-[M-2H]2-m/z 634.7071,found 634.6962.;
(3)将鸟苷二磷酸岩藻糖(GDP-Fuc)、软骨素六糖(VII)、MgCl2、Tris-HCl配制成水溶液,反应体系中GDP-Fuc的终浓度为40mM,MgCl2的终浓度为20mM,软骨素六糖(VII) 的终浓度为10mM,Tris-HCl的终浓度为50mM,反应体系pH为7.5,然后添加L-岩藻糖基转移酶Bfα1,3FT,反应48h,检测反应完成后,加入冰乙醇终止反应;将反应液离心10-20min,将上清通过Bio-Gel P2分子筛脱盐后,利用强碱性阴离子交换树脂介质分离纯化,再次利用 Bio-Gel P2分子筛脱盐后,得到产物XIV,其1H NMR、MS数据如下,
1H NMR(600MHz,D2O)δ5.40(d,J=4.0Hz,4H,H-1Fuc-A,H-1Fuc-B),5.22(d,J=3.7Hz,1H,H-1GalNAc-A),4.69(d,J=8.5Hz,1H,H-1GalNAc-A),4.57-4.44(m,10H,H-1GlcA-A,H-1GalNAc-B,H-1GlcA-B,H-1GalNAc-C,H-1GlcA-C),4.29(dd,J=10.7,3.9Hz,1H),4.22(d,J=2.1Hz,1H),4.16-4.12(m,4H),4.10-3.91(m,14H),3.84(s,4H),3.80-3.68(m,38H),3.60-3.56(m,8H),3.53-3.46(m,4H),3.33(t,J=8.2Hz,2H),2.07-2.01(m,18H,COOCH3,COOCH3,COOCH3),1.25(d,J=6.1Hz,12H,CH3 of Fucose-A,CH3 of Fucose-B).
ESI-MS(negative mode)calcd for C54H83N3O42 2-[M-2H]2-m/z 722.7231,found722.7092.;
(4)将鸟苷二磷酸岩藻糖(GDP-Fuc)、软骨素七糖(VIII)、MgCl2、Tris-HCl配制成水溶液,反应体系中GDP-Fuc的终浓度为60mM,MgCl2的终浓度为20mM,软骨素七糖 (VIII)的终浓度为10mM,Tris-HCl的终浓度为50mM,反应体系pH为7.5,然后添加 L-岩藻糖基转移酶Bfα1,3FT,反应48h,检测反应完成后,加入冰乙醇终止反应;将反应液离心10-20min,将上清通过Bio-Gel P2分子筛脱盐后,利用强碱性阴离子交换树脂介质分离纯化,再次利用Bio-Gel P2分子筛脱盐后,得到产物XV,其1H NMR、MS数据如下,
1H NMR(600MHz,D2O)δ5.36(d,J=3.3Hz,6H,H-1Fuc-A,H-1Fuc-B,H-1Fuc-C),5.18(d,J= 3.5Hz,1H),4.65(d,J=8.3Hz,1H,H-1GalNAc-A),4.53(d,J=8.1Hz,1H,H-1GalNAc-A),4.54-4.40 (m,12H,H-1GlcA-A,H-1GalNAc-B,H-1GlcA-B,H-1GalNAc-C,H-1GlcA-C,H-1GalNAc-D),4.25(d,J=11.6Hz, 1H),4.18(d,J=2.7Hz,1H),4.12-3.85(m,21H),3.80(m,6H),3.75-3.61(m,53H),3.55-3.52(m, 14H),2.04-1.99(m,24H,COOCH3,COOCH3,COOCH3,COOCH3),1.21(d,J=6.5Hz,18H, CH3 of Fucose-A,CH3 of Fucose-B,CH3 of Fucose-C).
ESI-MS(negative mode)calcd for C68H106N4O51 2-[M-2H]2-m/z 897.2917,found897.2679.。
实施例3岩藻糖基化软骨素寡糖的主链延伸
(1)20mM尿苷二磷酸N-乙酰氨基半乳糖(UDP-GalNAc)、10mM岩藻糖基化软骨素寡糖受体(XII)、20mM MgCl2、50mM Tris-HCl缓冲液(pH=7.5),1mg/mL软骨素合成酶的反应体系,30℃反应12h,检测反应完成后,加入冰乙醇终止反应;将反应液离心 10-20min,利用强碱性阴离子交换树脂介质分离纯化,得到产物XVI,其1H NMR、13C NMR 及MS数据如下,
1H NMR(600MHz,D2O)δ5.38(d,J=3.9Hz,2H,H-1Fuc),5.20(d,J=3.7Hz,1H, H-1GalNAc-A),4.66(d,J=8.5Hz,1H,H-1GalNAc-A),4.56-4.44(m,8H,H-1GlcA-A,H-1GalNAc-B, H-1GlcA -B,H-1GalNAc-C),4.27(dd,J=11.0,3.8Hz,1H),4.20(d,J=3.1Hz,1H),4.14-4.09(m,2H),4.07-4.02(m,3H),3.98-3.85(m,10H),3.83-3.66(m,35H),3.58-3.54(m,6H),3.37-3.33(m,2H), 2.05-1.98(m,18H,COOCH3,COOCH3,COOCH3),1.23(d,J=6.6Hz,6H,CH3 ofFucose).
13C NMR(151MHz,D2O)δ175.21,175.19,174.97,174.87,174.81,174.63,174.52,104.16, 103.97,103.82,100.87,99.69,98.21,95.08,91.09,80.66,79.47,77.62,76.54,76.27,75.14,74.87, 74.84,74.56,74.14,74.11,73.55,72.43,71.96,71.00,70.17,69.11,68.34,68.01,67.70,67.62, 67.53,66.57,61.49,61.18,60.95,52.22,50.77,48.74,22.42,22.37,22.13,21.91,15.27.
ESI-MS(negative mode)calcd for C42H66N3O32 -[M-H]-m/z1124.3635,found1124.3574; calcd for C42H65N3O32 2-[M-2H]2-m/z561.6781,found 561.6755.;
(2)20mM尿苷二磷酸N-乙酰氨基半乳糖(UDP-GalNAc)、10mM岩藻糖基化软骨素寡糖受体(XIV)、20mM MgCl2、50mM Tris-HCl缓冲液(pH=7.5),1mg/mL软骨素合成酶的反应体系,30℃反应12h,检测反应完成后,加入冰乙醇终止反应;将反应液离心10-20min,利用强碱性阴离子交换树脂介质分离纯化,得到产物XVII,其1H NMR、MS数据如下,
1H NMR(600MHz,D2O)δ5.40(d,J=3.7Hz,4H,H-1Fuc-A,H-1Fuc-B),5.22(d,J=3.7Hz,1H,H-1GalNAc-A),4.69(d,J=8.6Hz,1H,H-1GalNAc-A),4.59-4.46(m,12H,H-1GlcA-A,H-1GalNAc-B,H-1GlcA-B,H-1GalNAc-C,H-1GlcA-C,H-1GalNAc-D),4.29(dd,J=11.1,3.8Hz,1H),4.22(d,J=3.1Hz, 1H),4.15-3.88(m,24H),3.85-3.69(m,50H),3.61-3.54(m,10H),3.37(dd,J=9.5,7.9Hz,2H), 2.06-2.01(m,24H,COOCH3,COOCH3,COOCH3,COOCH3),1.25(d,J=6.6Hz,12H,CH3of Fucose-A,CH3 of Fucose-B).
ESI-MS(negative mode)calcd for C62H96N4O47 2-[M-2H]2-m/z824.2628,found824.2469.;
(3)20mM尿苷二磷酸葡糖醛酸(UDP-GlcA)、10mM岩藻糖基化软骨素寡糖受体(XVI)、20mM MgCl2、50mM Tris-HCl缓冲液(pH=7.5),1mg/mL软骨素合成酶的反应体系,30℃反应12h,检测反应完成后,加入冰乙醇终止反应;将反应液离心10-20min,利用强碱性阴离子交换树脂介质分离纯化,得到产物XVIII,其1H NMR、13C NMR及MS 数据如下,
1H NMR(600MHz,D2O)δ5.38(d,J=4.0Hz,2H,H-1Fuc),5.19(d,J=3.8Hz,1H, H-1GalNAc-A),4.66(d,J=8.5Hz,1H,H-1GalNAc-A),4.54-4.44(m,10H,H-1GlcA-A,H-1GalNAc-B, H-1GlcA-B,H-1GalNAc-C,H-1GlcA-C),4.26(dd,J=11.1,3.8Hz,1H),4.20-4.18(m,1H),4.15(d,J=3.1Hz,2H),4.13-4.08(m,2H),4.06-4.02(m,2H),4.02-3.89(m,9H),3.82-3.65(m,37H),3.58-3.53(m,6H),3.49-3.43(m,4H),3.36-3.28(m,4H),2.03-1.99(m,18H,COOCH3,COOCH3,COOCH3),1.23(d,J=6.6Hz,6H,CH3 of Fucose).
13C NMR(151MHz,D2O)δ175.79,175.14,175.13,174.87,174.85,174.80,174.50,174.46, 104.16,104.05,103.97,103.82,100.73,99.68,98.19,95.07,91.06,80.65,80.21,79.59,77.61, 76.49,76.44,76.18,76.17,75.99,75.17,74.88,74.83,74.55,74.12,73.55,72.60,72.37,71.94, 71.69,70.16,69.08,68.33,67.99,67.58,67.51,66.57,62.36,61.48,61.17,60.98,52.11,50.91, 50.76,48.72,22.39,22.11,21.88,15.25.
ESI-MS(negative mode)calcd for C48H74N3O38 -[M-H]-m/z1300.3956,found;calcd for C48H73N3O38 2-[M-2H]2-m/z649.6942,found 649.6843.;
(4)20mM尿苷二磷酸葡糖醛酸(UDP-GlcA)、10mM岩藻糖基化软骨素寡糖受体(XVIII)、20mM MgCl2、50mM Tris-HCl缓冲液(pH=7.5),1mg/mL软骨素合成酶的反应体系,30℃反应12h,检测反应完成后,加入冰乙醇终止反应;将反应液离心10-20min,利用强碱性阴离子交换树脂介质分离纯化,得到产物XIX,其1H NMR、13C NMR及MS数据如下,
1H NMR(600MHz,D2O)δ5.36(d,J=3.5Hz,2H,H-1Fuc),5.18(d,J=3.8Hz, 1H,H-1GalNAc-A),4.64(d,J=8.3Hz,1H,H-1GalNAc-A),4.54-4.42(m,12H,H-1GlcA-A,H-1GalNAc-B, H-1GlcA-B,H-1GalNAc-C,H-1GlcA-C,H-1GalNAc-D),4.25(dd,J=11.1,3.7Hz,1H),4.19-4.14(m,2H),4.12-4.08(m,4H),4.05-3.83(m,15H),3.80-3.65(m,46H),3.57-3.52(m,8H),3.35-3.31(m,4H), 2.03-1.98(m,24H,COOCH3,COOCH3,COOCH3,COOCH3),1.21(d,J=6.6Hz,6H,CH3 ofFucose).
13C NMR(151MHz,D2O)δ174.95,174.90,174.86,174.82,174.52,174.37,174.20,104.23, 104.17,100.92,100.77,99.71,98.21,95.09,91.09,80.66,80.60,80.23,79.63,79.46,77.61,76.44, 76.06,75.98,75.14,74.85,74.56,74.13,74.09,73.56,72.36,71.95,70.97,70.18,69.10,68.34, 68.01,67.73,67.69,67.64,67.51,66.58,62.37,61.48,61.18,60.96,60.94,52.23,52.14,50.89, 50.77,48.74,45.86,22.40,22.38,22.36,22.13,21.90,15.26.
ESI-MS(negative mode)calcd for C56H86N4O43 2-[M-2H]2-m/z751.2338,found751.2199.。
实施例4岩藻糖基化软骨素寡糖主链延伸后的按需岩藻糖基化修饰
(1)将鸟苷二磷酸岩藻糖(GDP-Fuc)、岩藻糖基化软骨素寡糖受体(XVI)、MgCl2、Tris-HCl配制成水溶液,反应体系中GDP-Fuc的终浓度为20mM、MgCl2的终浓度为20mM,岩藻糖基化软骨素寡糖(XVI)的终浓度为10mM,Tris-HCl的终浓度为50mM,反应体系 pH为7.5,然后添加L-岩藻糖基转移酶Hpα1,3FT S45F V30G,反应12h,检测反应完成后,加入冰乙醇终止反应;将反应液离心10-20min,将上清通过Bio-Gel P2分子筛脱盐后,利用强碱性阴离子交换树脂介质分离纯化,再次利用Bio-Gel P2分子筛脱盐后,得到产物XIII;
(2)将鸟苷二磷酸岩藻糖(GDP-Fuc)、岩藻糖基化软骨素寡糖受体(XVII)、MgCl2、Tris-HCl配制成水溶液,反应体系中GDP-Fuc的终浓度为20mM、MgCl2的终浓度为20mM,岩藻糖基化软骨素寡糖(XVII)的终浓度为10mM,Tris-HCl的终浓度为50mM,反应体系pH为7.5,然后添加L-岩藻糖基转移酶Hpα1,3FT S45F V30G,反应12h,检测反应完成后,加入冰乙醇终止反应;将反应液离心10-20min,将上清通过Bio-Gel P2分子筛脱盐后,利用强碱性阴离子交换树脂介质分离纯化,再次利用Bio-Gel P2分子筛脱盐后,得到产物XV;
(3)将鸟苷二磷酸岩藻糖(GDP-Fuc)、岩藻糖基化软骨素寡糖受体(XVIII)、MgCl2、Tris-HCl配制成水溶液,反应体系中GDP-Fuc的终浓度为20mM、MgCl2的终浓度为20mM,岩藻糖基化软骨素寡糖(XVIII)的终浓度为10mM,Tris-HCl的终浓度为50mM,反应体系pH为7.5,然后添加L-岩藻糖基转移酶Bfα1,3FT,检测反应完成后,加入冰乙醇终止反应;将反应液离心10-20min,将上清通过Bio-Gel P2分子筛脱盐后,利用强碱性阴离子交换树脂介质分离纯化,再次利用Bio-Gel P2分子筛脱盐后,得到产物XIV;
(4)将鸟苷二磷酸岩藻糖(GDP-Fuc)、岩藻糖基化软骨素寡糖受体(XIX)、MgCl2、Tris-HCl配制成水溶液,反应体系中GDP-Fuc的终浓度为40mM、MgCl2的终浓度为20mM,岩藻糖基化软骨素寡糖(XIX)的终浓度为10mM,Tris-HCl的终浓度为50mM,反应体系 pH为7.5,然后添加L-岩藻糖基转移酶Bfα1,3FT,检测反应完成后,加入冰乙醇终止反应;将反应液离心10-20min,将上清通过Bio-Gel P2分子筛脱盐后,利用强碱性阴离子交换树脂介质分离纯化,再次利用Bio-Gel P2分子筛脱盐后,得到产物XV;
(5)将鸟苷二磷酸岩藻糖(GDP-Fuc)、岩藻糖基化软骨素寡糖受体(XIX)、MgCl2、Tris-HCl配制成水溶液,反应体系中GDP-Fuc的终浓度为20mM、MgCl2的终浓度为20mM,岩藻糖基化软骨素寡糖(XIX)的终浓度为10mM,Tris-HCl的终浓度为50mM,反应体系 pH为7.5,然后添加L-岩藻糖基转移酶Hpα1,3FT S45F V30G,反应12h,检测反应完成后,加入冰乙醇终止反应;将反应液离心10-20min,将上清通过Bio-Gel P2分子筛脱盐后,利用强碱性阴离子交换树脂介质分离纯化,再次利用Bio-Gel P2分子筛脱盐后,得到产物XX,其1H NMR和MS数据如下,
1H NMR(600MHz,D2O)δ5.40(d,J=3.7Hz,4H,H-1Fuc-A,H-1Fuc-B),5.22(d,J=3.7Hz,1H, H-1GalNAc-A),4.69(d,J=8.6Hz,1H,H-1GalNAc-A),4.59-4.46(m,12H,H-1GlcA-A,H-1GalNAc-B,H-1GlcA-B,H-1GalNAc-C,H-1GlcA-C,H-1GalNAc-D),4.29(dd,J=11.1,3.8Hz,1H),4.22(d,J=3.1Hz, 1H),4.15-3.88(m,24H),3.85-3.69(m,50H),3.61-3.54(m,10H),3.37(dd,J=9.5,7.9Hz,2H), 2.06-2.01(m,24H,COOCH3,COOCH3,COOCH3,COOCH3),1.25(d,J=6.6Hz,12H,CH3of Fucose-A,CH3 of Fucose-B).
ESI-MS(negative mode)calcd for C62H96N4O47 2-[M-2H]2-m/z 824.2628,found824.2469.。
序列表
<110> 山东大学
<120> 一种岩藻糖基化软骨素及其制备方法和应用
<130> 1
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 996
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 1
atgtgtgatt gtctgagcat tattctgctg gttaaaatga agaagatcta cctgaagttc 60
gtggacttct gggacggctt tgataccatc agcaatttta tcgtggacgc cctgagcatc 120
cagtatgaag ttgttctgag caatgaaccg gattatctgt tttatagctg cttcggcacc 180
agccatctgg aatatgattg tattaaaatc atgtttatcg gcgagaacat cgtgccggat 240
ttcaatgttt gtgattacgc aattggcttc aactacatcg acttcggcga tcgttatctg 300
cgtctgccgc tgtatgcaat ttatgatggt tttagcaatc tgcagaacaa gaagatcgac 360
gttaataagg cactggatcg taagttctgt agcatcgttg ttagcaataa caagtgggca 420
gatccgattc gtgaaacctt ttttaaactg ctgagcagct ataagaaggt tgacagcggt 480
ggtcgtgcat ggaataatat tggtggtccg gttgataata agctggactt tattagccag 540
tacaagttca acatcgcctt cgagaacagc cgcgttctgg gttataccac cgaaaaaatt 600
atggagccga tgcaggttaa tagcatcccg gtttattggg gtaatccgct ggttggtaaa 660
gattttaatg tggacagctt cgtgaatgcc cacgactttg acagcctgga gcgtctggtt 720
gaatatatta ttgaactgga cagcagcaag gacaagtacc tggaaatgct ggagaagccg 780
tggctgctgg ataaaaccta tctggattgg aaacagctgc tgctgaattt tattaacaac 840
atcatgatga agagctacaa ggacgccaaa tacctggtta actacggtca tgcaggtaaa 900
tatcgtaatg agcagcgttt ttggggtcgt tgtgaacgta aatttaaact gcagcgtatt 960
atcgagtact acagccagct gtttgatcgc aaataa 996
<210> 2
<211> 1120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
atgtttcagc cgctgttgga tgcgtatgtg gaaagcgcga gcattgaaaa aatggcgagc 60
aaatctccgc cgccgctgaa aattgcgggg gcgaactggt ggggtgatga agaaatcaaa 120
gaattcaaaa acttcgtgct gtattttatt ctgagccagc gctataccat taccctgcat 180
cagaatccga acgaatttag cgatctggtg tttggcaatc cgctgggcag cgcgcgtaaa 240
attctgagct atcagaacgc gaaacgtgtg ttttataccg gcgaaaacga aagcccgaac 300
tttaacctgt ttgattatgc gatcggcttt gatgaactgg atttcaacga tcgttatctg 360
cgtatgccgc tgtattatga tcgtctgcat cataaagcgg aaagcgtgaa cgataccacc 420
gcgccgtata aactgaaaga taacagcctg tacgcgctga aaaaaccgag ccattgcttt 480
aaagaaaaac acccgaacct gtgcgcggtg gtgaacgatg aaagcgatcc gctgaaacgt 540
ggctttgcga gctttgtggc gagcaatccg aacgcgccga ttcgtaacgc gttttatgat 600
gcgctgaaca gcattgaacc ggtgaccggc ggtggcagcg tgcgtaacac cctgggctat 660
aacgtgaaaa acaaaaacga attcctgagc cagtataaat ttaacctgtg cttcgaaaac 720
acccagggct atggctatgt gaccgaaaaa atcatcgatg cgtatttcag ccataccatc 780
ccgatttatt ggggcagccc gagcgtggcg aaagatttca atccgaaaag cttcgtgaac 840
gtgcacgatt tcaaaaactt cgatgaagcg atcgattaca tcaaatatct gcatacccac 900
aaaaacgcgt atctggatat gctgtatgaa aatccgctga acaccctgga tggcaaagcg 960
tatttttatc agaacctgag cttcaaaaaa atcctggcct tcttcaaaac cattctggaa 1020
aacgatacca tctatcacga taatccgttt atcttttgcc gtgatctgaa cgaaccgctg 1080
gtgaccattg atgatctgcg tgttaactac ctcgagcacc 1120
<210> 3
<211> 331
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 3
Met Cys Asp Cys Leu Ser Ile Ile Leu Leu Val Lys Met Lys Lys Ile
1 5 10 15
Tyr Leu Lys Phe Val Asp Phe Trp Asp Gly Phe Asp Thr Ile Ser Asn
20 25 30
Phe Ile Val Asp Ala Leu Ser Ile Gln Tyr Glu Val Val Leu Ser Asn
35 40 45
Glu Pro Asp Tyr Leu Phe Tyr Ser Cys Phe Gly Thr Ser His Leu Glu
50 55 60
Tyr Asp Cys Ile Lys Ile Met Phe Ile Gly Glu Asn Ile Val Pro Asp
65 70 75 80
Phe Asn Val Cys Asp Tyr Ala Ile Gly Phe Asn Tyr Ile Asp Phe Gly
85 90 95
Asp Arg Tyr Leu Arg Leu Pro Leu Tyr Ala Ile Tyr Asp Gly Phe Ser
100 105 110
Asn Leu Gln Asn Lys Lys Ile Asp Val Asn Lys Ala Leu Asp Arg Lys
115 120 125
Phe Cys Ser Ile Val Val Ser Asn Asn Lys Trp Ala Asp Pro Ile Arg
130 135 140
Glu Thr Phe Phe Lys Leu Leu Ser Ser Tyr Lys Lys Val Asp Ser Gly
145 150 155 160
Gly Arg Ala Trp Asn Asn Ile Gly Gly Pro Val Asp Asn Lys Leu Asp
165 170 175
Phe Ile Ser Gln Tyr Lys Phe Asn Ile Ala Phe Glu Asn Ser Arg Val
180 185 190
Leu Gly Tyr Thr Thr Glu Lys Ile Met Glu Pro Met Gln Val Asn Ser
195 200 205
Ile Pro Val Tyr Trp Gly Asn Pro Leu Val Gly Lys Asp Phe Asn Val
210 215 220
Asp Ser Phe Val Asn Ala His Asp Phe Asp Ser Leu Glu Arg Leu Val
225 230 235 240
Glu Tyr Ile Ile Glu Leu Asp Ser Ser Lys Asp Lys Tyr Leu Glu Met
245 250 255
Leu Glu Lys Pro Trp Leu Leu Asp Lys Thr Tyr Leu Asp Trp Lys Gln
260 265 270
Leu Leu Leu Asn Phe Ile Asn Asn Ile Met Met Lys Ser Tyr Lys Asp
275 280 285
Ala Lys Tyr Leu Val Asn Tyr Gly His Ala Gly Lys Tyr Arg Asn Glu
290 295 300
Gln Arg Phe Trp Gly Arg Cys Glu Arg Lys Phe Lys Leu Gln Arg Ile
305 310 315 320
Ile Glu Tyr Tyr Ser Gln Leu Phe Asp Arg Lys
325 330
<210> 4
<211> 373
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 4
Met Phe Gln Pro Leu Leu Asp Ala Tyr Val Glu Ser Ala Ser Ile Glu
1 5 10 15
Lys Met Ala Ser Lys Ser Pro Pro Pro Leu Lys Ile Ala Gly Ala Asn
20 25 30
Trp Trp Gly Asp Glu Glu Ile Lys Glu Phe Lys Asn Phe Val Leu Tyr
35 40 45
Phe Ile Leu Ser Gln Arg Tyr Thr Ile Thr Leu His Gln Asn Pro Asn
50 55 60
Glu Phe Ser Asp Leu Val Phe Gly Asn Pro Leu Gly Ser Ala Arg Lys
65 70 75 80
Ile Leu Ser Tyr Gln Asn Ala Lys Arg Val Phe Tyr Thr Gly Glu Asn
85 90 95
Glu Ser Pro Asn Phe Asn Leu Phe Asp Tyr Ala Ile Gly Phe Asp Glu
100 105 110
Leu Asp Phe Asn Asp Arg Tyr Leu Arg Met Pro Leu Tyr Tyr Asp Arg
115 120 125
Leu His His Lys Ala Glu Ser Val Asn Asp Thr Thr Ala Pro Tyr Lys
130 135 140
Leu Lys Asp Asn Ser Leu Tyr Ala Leu Lys Lys Pro Ser His Cys Phe
145 150 155 160
Lys Glu Lys His Pro Asn Leu Cys Ala Val Val Asn Asp Glu Ser Asp
165 170 175
Pro Leu Lys Arg Gly Phe Ala Ser Phe Val Ala Ser Asn Pro Asn Ala
180 185 190
Pro Ile Arg Asn Ala Phe Tyr Asp Ala Leu Asn Ser Ile Glu Pro Val
195 200 205
Thr Gly Gly Gly Ser Val Arg Asn Thr Leu Gly Tyr Asn Val Lys Asn
210 215 220
Lys Asn Glu Phe Leu Ser Gln Tyr Lys Phe Asn Leu Cys Phe Glu Asn
225 230 235 240
Thr Gln Gly Tyr Gly Tyr Val Thr Glu Lys Ile Ile Asp Ala Tyr Phe
245 250 255
Ser His Thr Ile Pro Ile Tyr Trp Gly Ser Pro Ser Val Ala Lys Asp
260 265 270
Phe Asn Pro Lys Ser Phe Val Asn Val His Asp Phe Lys Asn Phe Asp
275 280 285
Glu Ala Ile Asp Tyr Ile Lys Tyr Leu His Thr His Lys Asn Ala Tyr
290 295 300
Leu Asp Met Leu Tyr Glu Asn Pro Leu Asn Thr Leu Asp Gly Lys Ala
305 310 315 320
Tyr Phe Tyr Gln Asn Leu Ser Phe Lys Lys Ile Leu Ala Phe Phe Lys
325 330 335
Thr Ile Leu Glu Asn Asp Thr Ile Tyr His Asp Asn Pro Phe Ile Phe
340 345 350
Cys Arg Asp Leu Asn Glu Pro Leu Val Thr Ile Asp Asp Leu Arg Val
355 360 365
Asn Tyr Leu Glu His
370
Claims (9)
1.一种岩藻糖基化软骨素的制备方法,所述的方法包括如下步骤:
将L-岩藻糖基转移酶与底物GDP-L-岩藻糖和软骨素寡糖混合,酶促反应,得到岩藻糖基化软骨素;
优选地,所述的L-岩藻糖基转移酶Bfα1,3FT的核苷酸序列如SEQ ID NO:1所示,氨基酸序列如SEQ ID NO:3所示;
优选地,所述的L-岩藻糖基转移酶Hpα1,3FT S45F V30G的核苷酸序列如SEQ ID NO:2所示,氨基酸序列如SEQ ID NO:4所示。
2.权利要求1所述的制备方法,其特征在于,所述制备方法还包括如下步骤:使用软骨素寡糖,在非还原端结构为D-GalNAc1-4-β-D-GlcA的GlcA O-3位使用L-岩藻糖基转移酶Hpα1,3FT S45F V30G催化进行单一位点的岩藻糖基化修饰。
3.权利要求1所述的制备方法,其特征在于,所述制备方法还包括如下步骤:使用软骨素寡糖,在D-GalNAc1-4-β-D-GlcA结构的所有GlcA O-3位使用L-岩藻糖基转移酶Bfα1,3FT催化进行岩藻糖基化修饰。
4.权利要求1所述的制备方法,其特征在于,所述制备方法还包括如下步骤:当岩藻糖基化软骨素寡糖的非还原端大于等于两个糖未岩藻糖基化修饰时,首先使用软骨素合成酶PmCS催化延伸岩藻糖基化软骨素的主链,然后在结构为D-GalNAc1-4-β-D-GlcA的GlcA O-3位使用L-岩藻糖基转移酶Bfα1,3FT或Hpα1,3FT S45F V30G催化其进一步进行岩藻糖基化修饰。
5.权利要求1所述的制备方法,其特征在于,所述软骨素寡糖中可岩藻糖基化的GlcA与GDP-L-岩藻糖的摩尔比为0.5-2:1-4。
8.权利要求7所述的制备方法,其特征在于,所述的n1为选自1-5的整数,优选地,所述的n1为选自1-3的整数。
9.一种岩藻糖基转移酶,其特征在于,所述岩藻糖基转移酶的核苷酸序列如SEQ IDNO:1或SEQ ID NO:2所示,氨基酸序列如SEQ ID NO:3或SEQ ID NO:4所示。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210763300.XA CN115058465A (zh) | 2022-06-30 | 2022-06-30 | 一种岩藻糖基化软骨素及其制备方法和应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210763300.XA CN115058465A (zh) | 2022-06-30 | 2022-06-30 | 一种岩藻糖基化软骨素及其制备方法和应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN115058465A true CN115058465A (zh) | 2022-09-16 |
Family
ID=83203919
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210763300.XA Pending CN115058465A (zh) | 2022-06-30 | 2022-06-30 | 一种岩藻糖基化软骨素及其制备方法和应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN115058465A (zh) |
Citations (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20040241645A1 (en) * | 1997-01-31 | 2004-12-02 | Genentech, Inc. | O-fucosyltransferase |
US6878819B1 (en) * | 1999-09-28 | 2005-04-12 | Jari Natunen | Fucosylated oligosaccharides and process for their preparation |
US20110053792A1 (en) * | 2007-10-08 | 2011-03-03 | Wolfgang Kemmner | Microarray for expression analysis of cellular glycosylation mechanism |
CN102329397A (zh) * | 2011-10-19 | 2012-01-25 | 中国科学院昆明植物研究所 | 一种岩藻糖化糖胺聚糖衍生物及其制备方法 |
CN103201380A (zh) * | 2010-10-11 | 2013-07-10 | 詹尼温生物技术有限责任公司 | 新岩藻糖基转移酶及它们的应用 |
CN108103039A (zh) * | 2016-11-25 | 2018-06-01 | 上海交通大学 | 一组岩藻糖基转移酶突变体及其筛选方法和应用 |
CN108794653A (zh) * | 2017-05-05 | 2018-11-13 | 北京大学 | 岩藻糖基化硫酸软骨素寡糖糖簇及其制备方法 |
CN110869508A (zh) * | 2017-07-07 | 2020-03-06 | 詹尼温生物技术有限责任公司 | 岩藻糖基转移酶及其在生产岩藻糖基化低聚糖中的用途 |
CN113056562A (zh) * | 2018-10-02 | 2021-06-29 | 齐米科技股份有限公司 | 使用底物输入体输出寡糖 |
CN113195509A (zh) * | 2018-12-18 | 2021-07-30 | 因比奥斯公司 | 生产3-岩藻糖基乳糖和转化乳糖的α-1,3-岩藻糖基转移酶 |
CN113528480A (zh) * | 2021-07-07 | 2021-10-22 | 上海交通大学 | 一种α-1,2-岩藻糖基转移酶突变体及其构建方法和应用 |
-
2022
- 2022-06-30 CN CN202210763300.XA patent/CN115058465A/zh active Pending
Patent Citations (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20040241645A1 (en) * | 1997-01-31 | 2004-12-02 | Genentech, Inc. | O-fucosyltransferase |
US6878819B1 (en) * | 1999-09-28 | 2005-04-12 | Jari Natunen | Fucosylated oligosaccharides and process for their preparation |
US20110053792A1 (en) * | 2007-10-08 | 2011-03-03 | Wolfgang Kemmner | Microarray for expression analysis of cellular glycosylation mechanism |
CN103201380A (zh) * | 2010-10-11 | 2013-07-10 | 詹尼温生物技术有限责任公司 | 新岩藻糖基转移酶及它们的应用 |
CN102329397A (zh) * | 2011-10-19 | 2012-01-25 | 中国科学院昆明植物研究所 | 一种岩藻糖化糖胺聚糖衍生物及其制备方法 |
CN108103039A (zh) * | 2016-11-25 | 2018-06-01 | 上海交通大学 | 一组岩藻糖基转移酶突变体及其筛选方法和应用 |
CN108794653A (zh) * | 2017-05-05 | 2018-11-13 | 北京大学 | 岩藻糖基化硫酸软骨素寡糖糖簇及其制备方法 |
CN110869508A (zh) * | 2017-07-07 | 2020-03-06 | 詹尼温生物技术有限责任公司 | 岩藻糖基转移酶及其在生产岩藻糖基化低聚糖中的用途 |
CN113056562A (zh) * | 2018-10-02 | 2021-06-29 | 齐米科技股份有限公司 | 使用底物输入体输出寡糖 |
CN113195509A (zh) * | 2018-12-18 | 2021-07-30 | 因比奥斯公司 | 生产3-岩藻糖基乳糖和转化乳糖的α-1,3-岩藻糖基转移酶 |
CN113528480A (zh) * | 2021-07-07 | 2021-10-22 | 上海交通大学 | 一种α-1,2-岩藻糖基转移酶突变体及其构建方法和应用 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Dumitriu | Polysaccharides in medicinal applications | |
Paulsen | Syntheses, conformations and X‐ray structure analyses of the saccharide chains from the core regions of glycoproteins | |
Nilsson | Enzymatic synthesis of oligosaccharides | |
US8193166B2 (en) | Epimerized derivatives of K5 polysaccharide with a very high degree of sulfation | |
WO1995024497A2 (en) | Enzymatic preparation of polysaccharides | |
JPH07501684A (ja) | 抗凝固剤およびその調製方法 | |
JP2004512013A (ja) | コンドロイチンシンターゼ遺伝子並びにその生成方法及び使用方法 | |
CN107312765B (zh) | 一种糖胺聚糖裂解酶及其编码基因与应用 | |
CN113603732B (zh) | 一种非动物源硫酸软骨素寡糖及其制备方法 | |
WO1998003524A1 (fr) | Nouveaux oligosaccharides a base de lactosamine et procede de preparation | |
Liu et al. | Glycan assembly strategy: from concept to application | |
Pan et al. | Highly convergent synthesis of a β-mannuronic acid alginate hexadecasaccharide | |
EP3897902B1 (en) | Separation of oligosaccharides | |
JPH01104089A (ja) | 二官能性オリゴ糖類およびそれから誘導される活性化合物類 | |
CN115058465A (zh) | 一种岩藻糖基化软骨素及其制备方法和应用 | |
US20040132142A1 (en) | Process for producing hyaluronic acid or its derivative | |
Cai et al. | Preparation and application of a ‘clickable’acceptor for enzymatic synthesis of heparin oligosaccharides | |
EP0839210B1 (en) | Method of producing derivatives of glc-beta 1-4glc-n-acetyl | |
WO1995018864A1 (en) | Method of producing derivatives of lactosamine | |
EP1801118A1 (en) | Process for synthesis of mucin-type peptides and muc1-related glycopeptides | |
JP3650409B2 (ja) | 低分子量分岐β−1,3−グルカン及び分岐ラミナリオリゴ糖の製造方法 | |
JP4768936B2 (ja) | 硫酸基を有するオリゴ糖 | |
JP5089998B2 (ja) | 4−硫酸化ヒアルロン酸 | |
JP4763997B2 (ja) | 構造明確なコンドロイチン4−硫酸及びその製造法 | |
Rosencrantz et al. | Chemo‐enzymatic cascade reactions for the synthesis of glycoconjugates |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination |