CN105969751A - 一种β-葡萄糖苷酶基因及其应用 - Google Patents
一种β-葡萄糖苷酶基因及其应用 Download PDFInfo
- Publication number
- CN105969751A CN105969751A CN201610424102.5A CN201610424102A CN105969751A CN 105969751 A CN105969751 A CN 105969751A CN 201610424102 A CN201610424102 A CN 201610424102A CN 105969751 A CN105969751 A CN 105969751A
- Authority
- CN
- China
- Prior art keywords
- gly
- ala
- val
- leu
- lys
- 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.)
- Granted
Links
- 108010047754 beta-Glucosidase Proteins 0.000 title claims abstract description 58
- 239000002773 nucleotide Substances 0.000 claims abstract description 8
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 8
- 210000004027 cell Anatomy 0.000 claims description 21
- 239000013598 vector Substances 0.000 claims description 10
- 241001198387 Escherichia coli BL21(DE3) Species 0.000 claims description 4
- 210000001236 prokaryotic cell Anatomy 0.000 claims description 4
- 102000006995 beta-Glucosidase Human genes 0.000 abstract description 27
- KDYFGRWQOYBRFD-UHFFFAOYSA-N succinic acid Chemical compound OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 abstract description 14
- 239000002689 soil Substances 0.000 abstract description 11
- 238000000034 method Methods 0.000 abstract description 9
- GUBGYTABKSRVRQ-CUHNMECISA-N D-Cellobiose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-CUHNMECISA-N 0.000 abstract description 7
- 239000001384 succinic acid Substances 0.000 abstract description 7
- 230000014509 gene expression Effects 0.000 abstract description 6
- 108010076504 Protein Sorting Signals Proteins 0.000 abstract description 4
- 102000004190 Enzymes Human genes 0.000 description 76
- 108090000790 Enzymes Proteins 0.000 description 76
- 229940088598 enzyme Drugs 0.000 description 76
- 108090000623 proteins and genes Proteins 0.000 description 30
- 239000012634 fragment Substances 0.000 description 29
- 238000006243 chemical reaction Methods 0.000 description 28
- 241000894006 Bacteria Species 0.000 description 25
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 17
- 239000007788 liquid Substances 0.000 description 16
- 239000000872 buffer Substances 0.000 description 12
- 230000000694 effects Effects 0.000 description 12
- 241000588724 Escherichia coli Species 0.000 description 11
- 239000000758 substrate Substances 0.000 description 11
- 230000009471 action Effects 0.000 description 10
- 238000010367 cloning Methods 0.000 description 10
- 239000013612 plasmid Substances 0.000 description 10
- 238000005516 engineering process Methods 0.000 description 9
- 239000000047 product Substances 0.000 description 9
- 238000012360 testing method Methods 0.000 description 9
- 239000000284 extract Substances 0.000 description 8
- 238000012163 sequencing technique Methods 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 108010073969 valyllysine Proteins 0.000 description 8
- 241000282326 Felis catus Species 0.000 description 7
- 108010077245 asparaginyl-proline Proteins 0.000 description 7
- 108010015792 glycyllysine Proteins 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 102000005744 Glycoside Hydrolases Human genes 0.000 description 6
- 108010031186 Glycoside Hydrolases Proteins 0.000 description 6
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 6
- 102000003960 Ligases Human genes 0.000 description 6
- 108090000364 Ligases Proteins 0.000 description 6
- 102000016943 Muramidase Human genes 0.000 description 6
- 108010014251 Muramidase Proteins 0.000 description 6
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 239000011543 agarose gel Substances 0.000 description 6
- 238000013467 fragmentation Methods 0.000 description 6
- 238000006062 fragmentation reaction Methods 0.000 description 6
- 229960000274 lysozyme Drugs 0.000 description 6
- 239000004325 lysozyme Substances 0.000 description 6
- 235000010335 lysozyme Nutrition 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 238000001816 cooling Methods 0.000 description 5
- 230000029087 digestion Effects 0.000 description 5
- VCSABYLVNWQYQE-UHFFFAOYSA-N Ala-Lys-Lys Natural products NCCCCC(NC(=O)C(N)C)C(=O)NC(CCCCN)C(O)=O VCSABYLVNWQYQE-UHFFFAOYSA-N 0.000 description 4
- BYKZWDGMJLNFJY-XKBZYTNZSA-N Gln-Ser-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(=O)N)N)O BYKZWDGMJLNFJY-XKBZYTNZSA-N 0.000 description 4
- 239000006137 Luria-Bertani broth Substances 0.000 description 4
- HAUUXTXKJNVIFY-ONGXEEELSA-N Lys-Gly-Val Chemical compound [H]N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O HAUUXTXKJNVIFY-ONGXEEELSA-N 0.000 description 4
- RBEATVHTWHTHTJ-KKUMJFAQSA-N Lys-Leu-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(O)=O RBEATVHTWHTHTJ-KKUMJFAQSA-N 0.000 description 4
- YDDDRTIPNTWGIG-SRVKXCTJSA-N Lys-Lys-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O YDDDRTIPNTWGIG-SRVKXCTJSA-N 0.000 description 4
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 4
- SITLTJHOQZFJGG-UHFFFAOYSA-N N-L-alpha-glutamyl-L-valine Natural products CC(C)C(C(O)=O)NC(=O)C(N)CCC(O)=O SITLTJHOQZFJGG-UHFFFAOYSA-N 0.000 description 4
- PZVGOVRNGKEFCB-KKHAAJSZSA-N Thr-Asn-Val Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](C(C)C)C(=O)O)N)O PZVGOVRNGKEFCB-KKHAAJSZSA-N 0.000 description 4
- 238000000246 agarose gel electrophoresis Methods 0.000 description 4
- 108010047495 alanylglycine Proteins 0.000 description 4
- 229960000723 ampicillin Drugs 0.000 description 4
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 4
- 230000003321 amplification Effects 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 108010043240 arginyl-leucyl-glycine Proteins 0.000 description 4
- 108010038633 aspartylglutamate Proteins 0.000 description 4
- 108010092854 aspartyllysine Proteins 0.000 description 4
- 238000006911 enzymatic reaction Methods 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 230000008676 import Effects 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 108010034529 leucyl-lysine Proteins 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 229910021645 metal ion Inorganic materials 0.000 description 4
- 238000003199 nucleic acid amplification method Methods 0.000 description 4
- 239000003960 organic solvent Substances 0.000 description 4
- 239000008363 phosphate buffer Substances 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 108010029020 prolylglycine Proteins 0.000 description 4
- 230000004044 response Effects 0.000 description 4
- 108010071207 serylmethionine Proteins 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 108010061238 threonyl-glycine Proteins 0.000 description 4
- 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 3
- 230000004544 DNA amplification Effects 0.000 description 3
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 3
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 3
- 108010005054 Deoxyribonuclease BamHI Proteins 0.000 description 3
- 108010042407 Endonucleases Proteins 0.000 description 3
- 102000004533 Endonucleases Human genes 0.000 description 3
- 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 3
- 239000006142 Luria-Bertani Agar Substances 0.000 description 3
- 239000001888 Peptone Substances 0.000 description 3
- 108010080698 Peptones Proteins 0.000 description 3
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 3
- 235000009392 Vitis Nutrition 0.000 description 3
- 241000219095 Vitis Species 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- BKHZIBWEHPHYAI-UHFFFAOYSA-N chloroform;3-methylbutan-1-ol Chemical compound ClC(Cl)Cl.CC(C)CCO BKHZIBWEHPHYAI-UHFFFAOYSA-N 0.000 description 3
- 230000004087 circulation Effects 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 230000001186 cumulative effect Effects 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 239000012160 loading buffer Substances 0.000 description 3
- 230000010355 oscillation Effects 0.000 description 3
- 235000019319 peptone Nutrition 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 235000018102 proteins Nutrition 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 238000010926 purge Methods 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 230000035939 shock Effects 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- BTJIUGUIPKRLHP-UHFFFAOYSA-N 4-nitrophenol Chemical compound OC1=CC=C([N+]([O-])=O)C=C1 BTJIUGUIPKRLHP-UHFFFAOYSA-N 0.000 description 2
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- CXRCVCURMBFFOL-FXQIFTODSA-N Ala-Ala-Pro Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(O)=O CXRCVCURMBFFOL-FXQIFTODSA-N 0.000 description 2
- JBVSSSZFNTXJDX-YTLHQDLWSA-N Ala-Ala-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](C)N JBVSSSZFNTXJDX-YTLHQDLWSA-N 0.000 description 2
- FXKNPWNXPQZLES-ZLUOBGJFSA-N Ala-Asn-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O FXKNPWNXPQZLES-ZLUOBGJFSA-N 0.000 description 2
- WCBVQNZTOKJWJS-ACZMJKKPSA-N Ala-Cys-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(O)=O)C(O)=O WCBVQNZTOKJWJS-ACZMJKKPSA-N 0.000 description 2
- CSAHOYQKNHGDHX-ACZMJKKPSA-N Ala-Gln-Asn Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O CSAHOYQKNHGDHX-ACZMJKKPSA-N 0.000 description 2
- OMMDTNGURYRDAC-NRPADANISA-N Ala-Glu-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O OMMDTNGURYRDAC-NRPADANISA-N 0.000 description 2
- BEMGNWZECGIJOI-WDSKDSINSA-N Ala-Gly-Glu Chemical compound [H]N[C@@H](C)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O BEMGNWZECGIJOI-WDSKDSINSA-N 0.000 description 2
- SIGTYDNEPYEXGK-ZANVPECISA-N Ala-Gly-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)CNC(=O)[C@@H](N)C)C(O)=O)=CNC2=C1 SIGTYDNEPYEXGK-ZANVPECISA-N 0.000 description 2
- LNNSWWRRYJLGNI-NAKRPEOUSA-N Ala-Ile-Val Chemical compound C[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C(C)C)C(O)=O LNNSWWRRYJLGNI-NAKRPEOUSA-N 0.000 description 2
- QUIGLPSHIFPEOV-CIUDSAMLSA-N Ala-Lys-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O QUIGLPSHIFPEOV-CIUDSAMLSA-N 0.000 description 2
- MDNAVFBZPROEHO-DCAQKATOSA-N Ala-Lys-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O MDNAVFBZPROEHO-DCAQKATOSA-N 0.000 description 2
- MDNAVFBZPROEHO-UHFFFAOYSA-N Ala-Lys-Val Natural products CC(C)C(C(O)=O)NC(=O)C(NC(=O)C(C)N)CCCCN MDNAVFBZPROEHO-UHFFFAOYSA-N 0.000 description 2
- RTZCUEHYUQZIDE-WHFBIAKZSA-N Ala-Ser-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O RTZCUEHYUQZIDE-WHFBIAKZSA-N 0.000 description 2
- YEBZNKPPOHFZJM-BPNCWPANSA-N Ala-Tyr-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C(C)C)C(O)=O YEBZNKPPOHFZJM-BPNCWPANSA-N 0.000 description 2
- ZCUFMRIQCPNOHZ-NRPADANISA-N Ala-Val-Gln Chemical compound C[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N ZCUFMRIQCPNOHZ-NRPADANISA-N 0.000 description 2
- XCIGOVDXZULBBV-DCAQKATOSA-N Ala-Val-Lys Chemical compound CC(C)[C@H](NC(=O)[C@H](C)N)C(=O)N[C@@H](CCCCN)C(O)=O XCIGOVDXZULBBV-DCAQKATOSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- HKRXJBBCQBAGIM-FXQIFTODSA-N Arg-Asp-Ser Chemical compound C(C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CO)C(=O)O)N)CN=C(N)N HKRXJBBCQBAGIM-FXQIFTODSA-N 0.000 description 2
- VNFWDYWTSHFRRG-SRVKXCTJSA-N Arg-Gln-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O VNFWDYWTSHFRRG-SRVKXCTJSA-N 0.000 description 2
- AUFHLLPVPSMEOG-YUMQZZPRSA-N Arg-Gly-Glu Chemical compound NC(N)=NCCC[C@H](N)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O AUFHLLPVPSMEOG-YUMQZZPRSA-N 0.000 description 2
- AGVNTAUPLWIQEN-ZPFDUUQYSA-N Arg-Ile-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N AGVNTAUPLWIQEN-ZPFDUUQYSA-N 0.000 description 2
- YBZMTKUDWXZLIX-UWVGGRQHSA-N Arg-Leu-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O YBZMTKUDWXZLIX-UWVGGRQHSA-N 0.000 description 2
- ZEBDYGZVMMKZNB-SRVKXCTJSA-N Arg-Met-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@H](CCCN=C(N)N)N ZEBDYGZVMMKZNB-SRVKXCTJSA-N 0.000 description 2
- ASQKVGRCKOFKIU-KZVJFYERSA-N Arg-Thr-Ala Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N)O ASQKVGRCKOFKIU-KZVJFYERSA-N 0.000 description 2
- OGZBJJLRKQZRHL-KJEVXHAQSA-N Arg-Thr-Tyr Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 OGZBJJLRKQZRHL-KJEVXHAQSA-N 0.000 description 2
- AZHXYLJRGVMQKW-UMPQAUOISA-N Arg-Trp-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)[C@H](CCCN=C(N)N)N)O AZHXYLJRGVMQKW-UMPQAUOISA-N 0.000 description 2
- VYZBPPBKFCHCIS-WPRPVWTQSA-N Arg-Val-Gly Chemical compound OC(=O)CNC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCN=C(N)N VYZBPPBKFCHCIS-WPRPVWTQSA-N 0.000 description 2
- SLKLLQWZQHXYSV-CIUDSAMLSA-N Asn-Ala-Lys Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O SLKLLQWZQHXYSV-CIUDSAMLSA-N 0.000 description 2
- BVLIJXXSXBUGEC-SRVKXCTJSA-N Asn-Asn-Tyr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O BVLIJXXSXBUGEC-SRVKXCTJSA-N 0.000 description 2
- QHBMKQWOIYJYMI-BYULHYEWSA-N Asn-Asn-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O QHBMKQWOIYJYMI-BYULHYEWSA-N 0.000 description 2
- VJTWLBMESLDOMK-WDSKDSINSA-N Asn-Gln-Gly Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O VJTWLBMESLDOMK-WDSKDSINSA-N 0.000 description 2
- UPALZCBCKAMGIY-PEFMBERDSA-N Asn-Gln-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O UPALZCBCKAMGIY-PEFMBERDSA-N 0.000 description 2
- RAQMSGVCGSJKCL-FOHZUACHSA-N Asn-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC(N)=O RAQMSGVCGSJKCL-FOHZUACHSA-N 0.000 description 2
- SEKBHZJLARBNPB-GHCJXIJMSA-N Asn-Ile-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O SEKBHZJLARBNPB-GHCJXIJMSA-N 0.000 description 2
- VOGCFWDZYYTEOY-DCAQKATOSA-N Asn-Lys-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(=O)N)N VOGCFWDZYYTEOY-DCAQKATOSA-N 0.000 description 2
- AYOAHKWVQLNPDM-HJGDQZAQSA-N Asn-Lys-Thr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O AYOAHKWVQLNPDM-HJGDQZAQSA-N 0.000 description 2
- KNENKKKUYGEZIO-FXQIFTODSA-N Asn-Met-Asn Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N KNENKKKUYGEZIO-FXQIFTODSA-N 0.000 description 2
- FMNBYVSGRCXWEK-FOHZUACHSA-N Asn-Thr-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O FMNBYVSGRCXWEK-FOHZUACHSA-N 0.000 description 2
- UXHYOWXTJLBEPG-GSSVUCPTSA-N Asn-Thr-Thr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O UXHYOWXTJLBEPG-GSSVUCPTSA-N 0.000 description 2
- WQAOZCVOOYUWKG-LSJOCFKGSA-N Asn-Val-Val Chemical compound CC(C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H](CC(=O)N)N WQAOZCVOOYUWKG-LSJOCFKGSA-N 0.000 description 2
- UWMIZBCTVWVMFI-FXQIFTODSA-N Asp-Ala-Arg Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O UWMIZBCTVWVMFI-FXQIFTODSA-N 0.000 description 2
- WSWYMRLTJVKRCE-ZLUOBGJFSA-N Asp-Ala-Asp Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(O)=O WSWYMRLTJVKRCE-ZLUOBGJFSA-N 0.000 description 2
- XEDQMTWEYFBOIK-ACZMJKKPSA-N Asp-Ala-Glu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O XEDQMTWEYFBOIK-ACZMJKKPSA-N 0.000 description 2
- OERMIMJQPQUIPK-FXQIFTODSA-N Asp-Arg-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(O)=O OERMIMJQPQUIPK-FXQIFTODSA-N 0.000 description 2
- DBWYWXNMZZYIRY-LPEHRKFASA-N Asp-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(=O)O)N)C(=O)O DBWYWXNMZZYIRY-LPEHRKFASA-N 0.000 description 2
- NYLBGYLHBDFRHL-VEVYYDQMSA-N Asp-Arg-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(O)=O NYLBGYLHBDFRHL-VEVYYDQMSA-N 0.000 description 2
- YNQIDCRRTWGHJD-ZLUOBGJFSA-N Asp-Asn-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC(O)=O YNQIDCRRTWGHJD-ZLUOBGJFSA-N 0.000 description 2
- FRSGNOZCTWDVFZ-ACZMJKKPSA-N Asp-Asp-Gln Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O FRSGNOZCTWDVFZ-ACZMJKKPSA-N 0.000 description 2
- CELPEWWLSXMVPH-CIUDSAMLSA-N Asp-Asp-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC(O)=O CELPEWWLSXMVPH-CIUDSAMLSA-N 0.000 description 2
- POTCZYQVVNXUIG-BQBZGAKWSA-N Asp-Gly-Pro Chemical compound OC(=O)C[C@H](N)C(=O)NCC(=O)N1CCC[C@H]1C(O)=O POTCZYQVVNXUIG-BQBZGAKWSA-N 0.000 description 2
- ILQCHXURSRRIRY-YUMQZZPRSA-N Asp-His-Gly Chemical compound C1=C(NC=N1)C[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CC(=O)O)N ILQCHXURSRRIRY-YUMQZZPRSA-N 0.000 description 2
- UBPMOJLRVMGTOQ-GARJFASQSA-N Asp-His-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CN=CN2)NC(=O)[C@H](CC(=O)O)N)C(=O)O UBPMOJLRVMGTOQ-GARJFASQSA-N 0.000 description 2
- AYFVRYXNDHBECD-YUMQZZPRSA-N Asp-Leu-Gly Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O AYFVRYXNDHBECD-YUMQZZPRSA-N 0.000 description 2
- LIVXPXUVXFRWNY-CIUDSAMLSA-N Asp-Lys-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O LIVXPXUVXFRWNY-CIUDSAMLSA-N 0.000 description 2
- JXGJJQJHXHXJQF-CIUDSAMLSA-N Asp-Met-Glu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(O)=O JXGJJQJHXHXJQF-CIUDSAMLSA-N 0.000 description 2
- YFGUZQQCSDZRBN-DCAQKATOSA-N Asp-Pro-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O YFGUZQQCSDZRBN-DCAQKATOSA-N 0.000 description 2
- MJJIHRWNWSQTOI-VEVYYDQMSA-N Asp-Thr-Arg Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O MJJIHRWNWSQTOI-VEVYYDQMSA-N 0.000 description 2
- PLNJUJGNLDSFOP-UWJYBYFXSA-N Asp-Tyr-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C)C(O)=O PLNJUJGNLDSFOP-UWJYBYFXSA-N 0.000 description 2
- VHUKCUHLFMRHOD-MELADBBJSA-N Asp-Tyr-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=C(C=C2)O)NC(=O)[C@H](CC(=O)O)N)C(=O)O VHUKCUHLFMRHOD-MELADBBJSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- YJIUYQKQBBQYHZ-ACZMJKKPSA-N Gln-Ala-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O YJIUYQKQBBQYHZ-ACZMJKKPSA-N 0.000 description 2
- OVQXQLWWJSNYFV-XEGUGMAKSA-N Gln-Ala-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CCC(N)=O)C)C(O)=O)=CNC2=C1 OVQXQLWWJSNYFV-XEGUGMAKSA-N 0.000 description 2
- KZEUVLLVULIPNX-GUBZILKMSA-N Gln-Asp-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCC(=O)N)N KZEUVLLVULIPNX-GUBZILKMSA-N 0.000 description 2
- ZQPOVSJFBBETHQ-CIUDSAMLSA-N Gln-Glu-Gln Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O ZQPOVSJFBBETHQ-CIUDSAMLSA-N 0.000 description 2
- DRDSQGHKTLSNEA-GLLZPBPUSA-N Gln-Glu-Thr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O DRDSQGHKTLSNEA-GLLZPBPUSA-N 0.000 description 2
- FGYPOQPQTUNESW-IUCAKERBSA-N Gln-Gly-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CCC(=O)N)N FGYPOQPQTUNESW-IUCAKERBSA-N 0.000 description 2
- VGTDBGYFVWOQTI-RYUDHWBXSA-N Gln-Gly-Phe Chemical compound NC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 VGTDBGYFVWOQTI-RYUDHWBXSA-N 0.000 description 2
- HYPVLWGNBIYTNA-GUBZILKMSA-N Gln-Leu-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O HYPVLWGNBIYTNA-GUBZILKMSA-N 0.000 description 2
- HPCOBEHVEHWREJ-DCAQKATOSA-N Gln-Lys-Glu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O HPCOBEHVEHWREJ-DCAQKATOSA-N 0.000 description 2
- JUUNNOLZGVYCJT-JYJNAYRXSA-N Gln-Phe-Lys Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)N)N JUUNNOLZGVYCJT-JYJNAYRXSA-N 0.000 description 2
- KVQOVQVGVKDZNW-GUBZILKMSA-N Gln-Ser-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(=O)N)N KVQOVQVGVKDZNW-GUBZILKMSA-N 0.000 description 2
- UEILCTONAMOGBR-RWRJDSDZSA-N Gln-Thr-Ile Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O UEILCTONAMOGBR-RWRJDSDZSA-N 0.000 description 2
- 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 2
- ATRHMOJQJWPVBQ-DRZSPHRISA-N Glu-Ala-Phe Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O ATRHMOJQJWPVBQ-DRZSPHRISA-N 0.000 description 2
- LTUVYLVIZHJCOQ-KKUMJFAQSA-N Glu-Arg-Phe Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O LTUVYLVIZHJCOQ-KKUMJFAQSA-N 0.000 description 2
- WOSRKEJQESVHGA-CIUDSAMLSA-N Glu-Arg-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(O)=O WOSRKEJQESVHGA-CIUDSAMLSA-N 0.000 description 2
- IQACOVZVOMVILH-FXQIFTODSA-N Glu-Glu-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O IQACOVZVOMVILH-FXQIFTODSA-N 0.000 description 2
- UHVIQGKBMXEVGN-WDSKDSINSA-N Glu-Gly-Asn Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O UHVIQGKBMXEVGN-WDSKDSINSA-N 0.000 description 2
- IVGJYOOGJLFKQE-AVGNSLFASA-N Glu-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)O)N IVGJYOOGJLFKQE-AVGNSLFASA-N 0.000 description 2
- RBXSZQRSEGYDFG-GUBZILKMSA-N Glu-Lys-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O RBXSZQRSEGYDFG-GUBZILKMSA-N 0.000 description 2
- YHOJJFFTSMWVGR-HJGDQZAQSA-N Glu-Met-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O YHOJJFFTSMWVGR-HJGDQZAQSA-N 0.000 description 2
- GMVCSRBOSIUTFC-FXQIFTODSA-N Glu-Ser-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(O)=O GMVCSRBOSIUTFC-FXQIFTODSA-N 0.000 description 2
- UMZHHILWZBFPGL-LOKLDPHHSA-N Glu-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)O)N)O UMZHHILWZBFPGL-LOKLDPHHSA-N 0.000 description 2
- CAQXJMUDOLSBPF-SUSMZKCASA-N Glu-Thr-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O CAQXJMUDOLSBPF-SUSMZKCASA-N 0.000 description 2
- NTHIHAUEXVTXQG-KKUMJFAQSA-N Glu-Tyr-Arg Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](CCC(=O)O)N)O NTHIHAUEXVTXQG-KKUMJFAQSA-N 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- BRFJMRSRMOMIMU-WHFBIAKZSA-N Gly-Ala-Asn Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(O)=O BRFJMRSRMOMIMU-WHFBIAKZSA-N 0.000 description 2
- YMUFWNJHVPQNQD-ZKWXMUAHSA-N Gly-Ala-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)CN YMUFWNJHVPQNQD-ZKWXMUAHSA-N 0.000 description 2
- PHONXOACARQMPM-BQBZGAKWSA-N Gly-Ala-Met Chemical compound [H]NCC(=O)N[C@@H](C)C(=O)N[C@@H](CCSC)C(O)=O PHONXOACARQMPM-BQBZGAKWSA-N 0.000 description 2
- XBWMTPAIUQIWKA-BYULHYEWSA-N Gly-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)CN XBWMTPAIUQIWKA-BYULHYEWSA-N 0.000 description 2
- FZQLXNIMCPJVJE-YUMQZZPRSA-N Gly-Asp-Leu Chemical compound [H]NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O FZQLXNIMCPJVJE-YUMQZZPRSA-N 0.000 description 2
- KTSZUNRRYXPZTK-BQBZGAKWSA-N Gly-Gln-Glu Chemical compound NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O KTSZUNRRYXPZTK-BQBZGAKWSA-N 0.000 description 2
- QSVCIFZPGLOZGH-WDSKDSINSA-N Gly-Glu-Ser Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O QSVCIFZPGLOZGH-WDSKDSINSA-N 0.000 description 2
- QPCVIQJVRGXUSA-LURJTMIESA-N Gly-Gly-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)CNC(=O)CN QPCVIQJVRGXUSA-LURJTMIESA-N 0.000 description 2
- YWAQATDNEKZFFK-BYPYZUCNSA-N Gly-Gly-Ser Chemical compound NCC(=O)NCC(=O)N[C@@H](CO)C(O)=O YWAQATDNEKZFFK-BYPYZUCNSA-N 0.000 description 2
- ADZGCWWDPFDHCY-ZETCQYMHSA-N Gly-His-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)CN)CC1=CN=CN1 ADZGCWWDPFDHCY-ZETCQYMHSA-N 0.000 description 2
- SXJHOPPTOJACOA-QXEWZRGKSA-N Gly-Ile-Arg Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CCCN=C(N)N SXJHOPPTOJACOA-QXEWZRGKSA-N 0.000 description 2
- OQQKUTVULYLCDG-ONGXEEELSA-N Gly-Lys-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CCCCN)NC(=O)CN)C(O)=O OQQKUTVULYLCDG-ONGXEEELSA-N 0.000 description 2
- OJNZVYSGVYLQIN-BQBZGAKWSA-N Gly-Met-Asp Chemical compound [H]NCC(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(O)=O OJNZVYSGVYLQIN-BQBZGAKWSA-N 0.000 description 2
- VDCRBJACQKOSMS-JSGCOSHPSA-N Gly-Phe-Val Chemical compound [H]NCC(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O VDCRBJACQKOSMS-JSGCOSHPSA-N 0.000 description 2
- NVTPVQLIZCOJFK-FOHZUACHSA-N Gly-Thr-Asp Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(O)=O NVTPVQLIZCOJFK-FOHZUACHSA-N 0.000 description 2
- MYXNLWDWWOTERK-BHNWBGBOSA-N Gly-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)CN)O MYXNLWDWWOTERK-BHNWBGBOSA-N 0.000 description 2
- GWNIGUKSRJBIHX-STQMWFEESA-N Gly-Tyr-Arg Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)CN)O GWNIGUKSRJBIHX-STQMWFEESA-N 0.000 description 2
- UVTSZKIATYSKIR-RYUDHWBXSA-N Gly-Tyr-Glu Chemical compound [H]NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O UVTSZKIATYSKIR-RYUDHWBXSA-N 0.000 description 2
- DNAZKGFYFRGZIH-QWRGUYRKSA-N Gly-Tyr-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CC=C(O)C=C1 DNAZKGFYFRGZIH-QWRGUYRKSA-N 0.000 description 2
- DUAWRXXTOQOECJ-JSGCOSHPSA-N Gly-Tyr-Val Chemical compound [H]NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C(C)C)C(O)=O DUAWRXXTOQOECJ-JSGCOSHPSA-N 0.000 description 2
- GJHWILMUOANXTG-WPRPVWTQSA-N Gly-Val-Arg Chemical compound [H]NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O GJHWILMUOANXTG-WPRPVWTQSA-N 0.000 description 2
- RVKIPWVMZANZLI-UHFFFAOYSA-N H-Lys-Trp-OH Natural products C1=CC=C2C(CC(NC(=O)C(N)CCCCN)C(O)=O)=CNC2=C1 RVKIPWVMZANZLI-UHFFFAOYSA-N 0.000 description 2
- DAKSMIWQZPHRIB-BZSNNMDCSA-N His-Tyr-Leu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(O)=O DAKSMIWQZPHRIB-BZSNNMDCSA-N 0.000 description 2
- JXUGDUWBMKIJDC-NAKRPEOUSA-N Ile-Ala-Arg Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O JXUGDUWBMKIJDC-NAKRPEOUSA-N 0.000 description 2
- LQSBBHNVAVNZSX-GHCJXIJMSA-N Ile-Ala-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](CC(=O)N)C(=O)O)N LQSBBHNVAVNZSX-GHCJXIJMSA-N 0.000 description 2
- ATXGFMOBVKSOMK-PEDHHIEDSA-N Ile-Arg-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N ATXGFMOBVKSOMK-PEDHHIEDSA-N 0.000 description 2
- OVPYIUNCVSOVNF-KQXIARHKSA-N Ile-Gln-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N1CCC[C@@H]1C(=O)O)N OVPYIUNCVSOVNF-KQXIARHKSA-N 0.000 description 2
- OVPYIUNCVSOVNF-ZPFDUUQYSA-N Ile-Gln-Pro Natural products CC[C@H](C)[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N1CCC[C@H]1C(O)=O OVPYIUNCVSOVNF-ZPFDUUQYSA-N 0.000 description 2
- GQKSJYINYYWPMR-NGZCFLSTSA-N Ile-Gly-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N1CCC[C@@H]1C(=O)O)N GQKSJYINYYWPMR-NGZCFLSTSA-N 0.000 description 2
- XQLGNKLSPYCRMZ-HJWJTTGWSA-N Ile-Phe-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(=O)O)N XQLGNKLSPYCRMZ-HJWJTTGWSA-N 0.000 description 2
- AKQFLPNANHNTLP-VKOGCVSHSA-N Ile-Pro-Trp Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC2=CNC3=CC=CC=C32)C(=O)O)N AKQFLPNANHNTLP-VKOGCVSHSA-N 0.000 description 2
- YKZAMJXNJUWFIK-JBDRJPRFSA-N Ile-Ser-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(=O)O)N YKZAMJXNJUWFIK-JBDRJPRFSA-N 0.000 description 2
- REXAUQBGSGDEJY-IGISWZIWSA-N Ile-Tyr-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N REXAUQBGSGDEJY-IGISWZIWSA-N 0.000 description 2
- PMGDADKJMCOXHX-UHFFFAOYSA-N L-Arginyl-L-glutamin-acetat Natural products NC(=N)NCCCC(N)C(=O)NC(CCC(N)=O)C(O)=O PMGDADKJMCOXHX-UHFFFAOYSA-N 0.000 description 2
- 241000880493 Leptailurus serval Species 0.000 description 2
- OIARJGNVARWKFP-YUMQZZPRSA-N Leu-Asn-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O OIARJGNVARWKFP-YUMQZZPRSA-N 0.000 description 2
- DLCOFDAHNMMQPP-SRVKXCTJSA-N Leu-Asp-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O DLCOFDAHNMMQPP-SRVKXCTJSA-N 0.000 description 2
- MMEDVBWCMGRKKC-GARJFASQSA-N Leu-Asp-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N1CCC[C@@H]1C(=O)O)N MMEDVBWCMGRKKC-GARJFASQSA-N 0.000 description 2
- DLCXCECTCPKKCD-GUBZILKMSA-N Leu-Gln-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O DLCXCECTCPKKCD-GUBZILKMSA-N 0.000 description 2
- GPICTNQYKHHHTH-GUBZILKMSA-N Leu-Gln-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O GPICTNQYKHHHTH-GUBZILKMSA-N 0.000 description 2
- WIDZHJTYKYBLSR-DCAQKATOSA-N Leu-Glu-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O WIDZHJTYKYBLSR-DCAQKATOSA-N 0.000 description 2
- LAPSXOAUPNOINL-YUMQZZPRSA-N Leu-Gly-Asp Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC(O)=O LAPSXOAUPNOINL-YUMQZZPRSA-N 0.000 description 2
- CCQLQKZTXZBXTN-NHCYSSNCSA-N Leu-Gly-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H]([C@@H](C)CC)C(O)=O CCQLQKZTXZBXTN-NHCYSSNCSA-N 0.000 description 2
- KEVYYIMVELOXCT-KBPBESRZSA-N Leu-Gly-Phe Chemical compound CC(C)C[C@H]([NH3+])C(=O)NCC(=O)N[C@H](C([O-])=O)CC1=CC=CC=C1 KEVYYIMVELOXCT-KBPBESRZSA-N 0.000 description 2
- YFBBUHJJUXXZOF-UWVGGRQHSA-N Leu-Gly-Pro Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N1CCC[C@H]1C(O)=O YFBBUHJJUXXZOF-UWVGGRQHSA-N 0.000 description 2
- KXODZBLFVFSLAI-AVGNSLFASA-N Leu-His-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC(C)C)CC1=CN=CN1 KXODZBLFVFSLAI-AVGNSLFASA-N 0.000 description 2
- HRTRLSRYZZKPCO-BJDJZHNGSA-N Leu-Ile-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O HRTRLSRYZZKPCO-BJDJZHNGSA-N 0.000 description 2
- PDQDCFBVYXEFSD-SRVKXCTJSA-N Leu-Leu-Asp Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O PDQDCFBVYXEFSD-SRVKXCTJSA-N 0.000 description 2
- JNDYEOUZBLOVOF-AVGNSLFASA-N Leu-Leu-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O JNDYEOUZBLOVOF-AVGNSLFASA-N 0.000 description 2
- LXKNSJLSGPNHSK-KKUMJFAQSA-N Leu-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N LXKNSJLSGPNHSK-KKUMJFAQSA-N 0.000 description 2
- IEWBEPKLKUXQBU-VOAKCMCISA-N Leu-Leu-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O IEWBEPKLKUXQBU-VOAKCMCISA-N 0.000 description 2
- LZHJZLHSRGWBBE-IHRRRGAJSA-N Leu-Lys-Val Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O LZHJZLHSRGWBBE-IHRRRGAJSA-N 0.000 description 2
- WMIOEVKKYIMVKI-DCAQKATOSA-N Leu-Pro-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O WMIOEVKKYIMVKI-DCAQKATOSA-N 0.000 description 2
- RRVCZCNFXIFGRA-DCAQKATOSA-N Leu-Pro-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(O)=O RRVCZCNFXIFGRA-DCAQKATOSA-N 0.000 description 2
- KWLWZYMNUZJKMZ-IHRRRGAJSA-N Leu-Pro-Leu Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O KWLWZYMNUZJKMZ-IHRRRGAJSA-N 0.000 description 2
- JDBQSGMJBMPNFT-AVGNSLFASA-N Leu-Pro-Val Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O JDBQSGMJBMPNFT-AVGNSLFASA-N 0.000 description 2
- PPGBXYKMUMHFBF-KATARQTJSA-N Leu-Ser-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PPGBXYKMUMHFBF-KATARQTJSA-N 0.000 description 2
- SQUFDMCWMFOEBA-KKUMJFAQSA-N Leu-Ser-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 SQUFDMCWMFOEBA-KKUMJFAQSA-N 0.000 description 2
- ICYRCNICGBJLGM-HJGDQZAQSA-N Leu-Thr-Asp Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(O)=O ICYRCNICGBJLGM-HJGDQZAQSA-N 0.000 description 2
- VDIARPPNADFEAV-WEDXCCLWSA-N Leu-Thr-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O VDIARPPNADFEAV-WEDXCCLWSA-N 0.000 description 2
- JGKHAFUAPZCCDU-BZSNNMDCSA-N Leu-Tyr-Leu Chemical compound CC(C)C[C@H]([NH3+])C(=O)N[C@H](C(=O)N[C@@H](CC(C)C)C([O-])=O)CC1=CC=C(O)C=C1 JGKHAFUAPZCCDU-BZSNNMDCSA-N 0.000 description 2
- FBNPMTNBFFAMMH-UHFFFAOYSA-N Leu-Val-Arg Natural products CC(C)CC(N)C(=O)NC(C(C)C)C(=O)NC(C(O)=O)CCCN=C(N)N FBNPMTNBFFAMMH-UHFFFAOYSA-N 0.000 description 2
- 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 2
- MVJRBCJCRYGCKV-GVXVVHGQSA-N Leu-Val-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O MVJRBCJCRYGCKV-GVXVVHGQSA-N 0.000 description 2
- QESXLSQLQHHTIX-RHYQMDGZSA-N Leu-Val-Thr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O QESXLSQLQHHTIX-RHYQMDGZSA-N 0.000 description 2
- KCXUCYYZNZFGLL-SRVKXCTJSA-N Lys-Ala-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O KCXUCYYZNZFGLL-SRVKXCTJSA-N 0.000 description 2
- WSXTWLJHTLRFLW-SRVKXCTJSA-N Lys-Ala-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O WSXTWLJHTLRFLW-SRVKXCTJSA-N 0.000 description 2
- YRWCPXOFBKTCFY-NUTKFTJISA-N Lys-Ala-Trp Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](CCCCN)N YRWCPXOFBKTCFY-NUTKFTJISA-N 0.000 description 2
- VHNOAIFVYUQOOY-XUXIUFHCSA-N Lys-Arg-Ile Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O VHNOAIFVYUQOOY-XUXIUFHCSA-N 0.000 description 2
- AAORVPFVUIHEAB-YUMQZZPRSA-N Lys-Asp-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O AAORVPFVUIHEAB-YUMQZZPRSA-N 0.000 description 2
- LXNPMPIQDNSMTA-AVGNSLFASA-N Lys-Gln-His Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(O)=O)CC1=CN=CN1 LXNPMPIQDNSMTA-AVGNSLFASA-N 0.000 description 2
- IRRZDAIFYHNIIN-JYJNAYRXSA-N Lys-Gln-Tyr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O IRRZDAIFYHNIIN-JYJNAYRXSA-N 0.000 description 2
- VQXAVLQBQJMENB-SRVKXCTJSA-N Lys-Glu-Met Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCSC)C(O)=O VQXAVLQBQJMENB-SRVKXCTJSA-N 0.000 description 2
- GQFDWEDHOQRNLC-QWRGUYRKSA-N Lys-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN GQFDWEDHOQRNLC-QWRGUYRKSA-N 0.000 description 2
- FGMHXLULNHTPID-KKUMJFAQSA-N Lys-His-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CCCCN)C(O)=O)CC1=CN=CN1 FGMHXLULNHTPID-KKUMJFAQSA-N 0.000 description 2
- IUWMQCZOTYRXPL-ZPFDUUQYSA-N Lys-Ile-Asp Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(O)=O)C(O)=O IUWMQCZOTYRXPL-ZPFDUUQYSA-N 0.000 description 2
- MYZMQWHPDAYKIE-SRVKXCTJSA-N Lys-Leu-Ala Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O MYZMQWHPDAYKIE-SRVKXCTJSA-N 0.000 description 2
- WVJNGSFKBKOKRV-AJNGGQMLSA-N Lys-Leu-Ile Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WVJNGSFKBKOKRV-AJNGGQMLSA-N 0.000 description 2
- YPLVCBKEPJPBDQ-MELADBBJSA-N Lys-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCCCN)N YPLVCBKEPJPBDQ-MELADBBJSA-N 0.000 description 2
- XOQMURBBIXRRCR-SRVKXCTJSA-N Lys-Lys-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CCCCN XOQMURBBIXRRCR-SRVKXCTJSA-N 0.000 description 2
- RIJCHEVHFWMDKD-SRVKXCTJSA-N Lys-Lys-Asn Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O RIJCHEVHFWMDKD-SRVKXCTJSA-N 0.000 description 2
- 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 2
- 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 2
- SBQDRNOLGSYHQA-YUMQZZPRSA-N Lys-Ser-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)NCC(O)=O SBQDRNOLGSYHQA-YUMQZZPRSA-N 0.000 description 2
- MIFFFXHMAHFACR-KATARQTJSA-N Lys-Ser-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CCCCN MIFFFXHMAHFACR-KATARQTJSA-N 0.000 description 2
- MDDUIRLQCYVRDO-NHCYSSNCSA-N Lys-Val-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCCN MDDUIRLQCYVRDO-NHCYSSNCSA-N 0.000 description 2
- OZVXDDFYCQOPFD-XQQFMLRXSA-N Lys-Val-Pro Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCCCN)N OZVXDDFYCQOPFD-XQQFMLRXSA-N 0.000 description 2
- GILLQRYAWOMHED-DCAQKATOSA-N Lys-Val-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCCN GILLQRYAWOMHED-DCAQKATOSA-N 0.000 description 2
- HUKLXYYPZWPXCC-KZVJFYERSA-N Met-Ala-Thr Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O HUKLXYYPZWPXCC-KZVJFYERSA-N 0.000 description 2
- FRWZTWWOORIIBA-FXQIFTODSA-N Met-Asn-Asn Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(=O)N)C(=O)O)N FRWZTWWOORIIBA-FXQIFTODSA-N 0.000 description 2
- MCNGIXXCMJAURZ-VEVYYDQMSA-N Met-Asp-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCSC)N)O MCNGIXXCMJAURZ-VEVYYDQMSA-N 0.000 description 2
- KQBJYJXPZBNEIK-DCAQKATOSA-N Met-Glu-Arg Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CCCNC(N)=N KQBJYJXPZBNEIK-DCAQKATOSA-N 0.000 description 2
- YCUSPBPZVJDMII-YUMQZZPRSA-N Met-Gly-Glu Chemical compound CSCC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCC(O)=O YCUSPBPZVJDMII-YUMQZZPRSA-N 0.000 description 2
- ILKCLLLOGPDNIP-RCWTZXSCSA-N Met-Met-Thr Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O ILKCLLLOGPDNIP-RCWTZXSCSA-N 0.000 description 2
- JQHYVIKEFYETEW-IHRRRGAJSA-N Met-Phe-Ser Chemical compound CSCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CO)C(O)=O)CC1=CC=CC=C1 JQHYVIKEFYETEW-IHRRRGAJSA-N 0.000 description 2
- VSJAPSMRFYUOKS-IUCAKERBSA-N Met-Pro-Gly Chemical compound CSCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O VSJAPSMRFYUOKS-IUCAKERBSA-N 0.000 description 2
- LXCSZPUQKMTXNW-BQBZGAKWSA-N Met-Ser-Gly Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O LXCSZPUQKMTXNW-BQBZGAKWSA-N 0.000 description 2
- QAVZUKIPOMBLMC-AVGNSLFASA-N Met-Val-Leu Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CC(C)C QAVZUKIPOMBLMC-AVGNSLFASA-N 0.000 description 2
- 108010047562 NGR peptide Proteins 0.000 description 2
- FPTXMUIBLMGTQH-ONGXEEELSA-N Phe-Ala-Gly Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=CC=C1 FPTXMUIBLMGTQH-ONGXEEELSA-N 0.000 description 2
- JWQWPTLEOFNCGX-AVGNSLFASA-N Phe-Glu-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 JWQWPTLEOFNCGX-AVGNSLFASA-N 0.000 description 2
- MMJJFXWMCMJMQA-STQMWFEESA-N Phe-Pro-Gly Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)NCC(O)=O)C1=CC=CC=C1 MMJJFXWMCMJMQA-STQMWFEESA-N 0.000 description 2
- GLJZDMZJHFXJQG-BZSNNMDCSA-N Phe-Ser-Phe Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O GLJZDMZJHFXJQG-BZSNNMDCSA-N 0.000 description 2
- 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 2
- APKRGYLBSCWJJP-FXQIFTODSA-N Pro-Ala-Asp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(O)=O APKRGYLBSCWJJP-FXQIFTODSA-N 0.000 description 2
- IFMDQWDAJUMMJC-DCAQKATOSA-N Pro-Ala-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O IFMDQWDAJUMMJC-DCAQKATOSA-N 0.000 description 2
- HFZNNDWPHBRNPV-KZVJFYERSA-N Pro-Ala-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O HFZNNDWPHBRNPV-KZVJFYERSA-N 0.000 description 2
- SWXSLPHTJVAWDF-VEVYYDQMSA-N Pro-Asn-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SWXSLPHTJVAWDF-VEVYYDQMSA-N 0.000 description 2
- XUSDDSLCRPUKLP-QXEWZRGKSA-N Pro-Asp-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H]1CCCN1 XUSDDSLCRPUKLP-QXEWZRGKSA-N 0.000 description 2
- VOZIBWWZSBIXQN-SRVKXCTJSA-N Pro-Glu-Lys Chemical compound NCCCC[C@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H]1CCCN1)C(O)=O VOZIBWWZSBIXQN-SRVKXCTJSA-N 0.000 description 2
- IBGCFJDLCYTKPW-NAKRPEOUSA-N Pro-Ile-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H]1CCCN1 IBGCFJDLCYTKPW-NAKRPEOUSA-N 0.000 description 2
- VZKBJNBZMZHKRC-XUXIUFHCSA-N Pro-Ile-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O VZKBJNBZMZHKRC-XUXIUFHCSA-N 0.000 description 2
- RYJRPPUATSKNAY-STECZYCISA-N Pro-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]2CCCN2 RYJRPPUATSKNAY-STECZYCISA-N 0.000 description 2
- HFNPOYOKIPGAEI-SRVKXCTJSA-N Pro-Leu-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H]1CCCN1 HFNPOYOKIPGAEI-SRVKXCTJSA-N 0.000 description 2
- WOIFYRZPIORBRY-AVGNSLFASA-N Pro-Lys-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O WOIFYRZPIORBRY-AVGNSLFASA-N 0.000 description 2
- JIWJRKNYLSHONY-KKUMJFAQSA-N Pro-Phe-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O JIWJRKNYLSHONY-KKUMJFAQSA-N 0.000 description 2
- POQFNPILEQEODH-FXQIFTODSA-N Pro-Ser-Ala Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O POQFNPILEQEODH-FXQIFTODSA-N 0.000 description 2
- GZNYIXWOIUFLGO-ZJDVBMNYSA-N Pro-Thr-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GZNYIXWOIUFLGO-ZJDVBMNYSA-N 0.000 description 2
- VVAWNPIOYXAMAL-KJEVXHAQSA-N Pro-Thr-Tyr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O VVAWNPIOYXAMAL-KJEVXHAQSA-N 0.000 description 2
- KHRLUIPIMIQFGT-AVGNSLFASA-N Pro-Val-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O KHRLUIPIMIQFGT-AVGNSLFASA-N 0.000 description 2
- FIODMZKLZFLYQP-GUBZILKMSA-N Pro-Val-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O FIODMZKLZFLYQP-GUBZILKMSA-N 0.000 description 2
- 229920002684 Sepharose Polymers 0.000 description 2
- SRTCFKGBYBZRHA-ACZMJKKPSA-N Ser-Ala-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O SRTCFKGBYBZRHA-ACZMJKKPSA-N 0.000 description 2
- FCRMLGJMPXCAHD-FXQIFTODSA-N Ser-Arg-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(O)=O FCRMLGJMPXCAHD-FXQIFTODSA-N 0.000 description 2
- OYEDZGNMSBZCIM-XGEHTFHBSA-N Ser-Arg-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OYEDZGNMSBZCIM-XGEHTFHBSA-N 0.000 description 2
- HEQPKICPPDOSIN-SRVKXCTJSA-N Ser-Asp-Tyr Chemical compound OC[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 HEQPKICPPDOSIN-SRVKXCTJSA-N 0.000 description 2
- ULVMNZOKDBHKKI-ACZMJKKPSA-N Ser-Gln-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O ULVMNZOKDBHKKI-ACZMJKKPSA-N 0.000 description 2
- MIJWOJAXARLEHA-WDSKDSINSA-N Ser-Gly-Glu Chemical compound OC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCC(O)=O MIJWOJAXARLEHA-WDSKDSINSA-N 0.000 description 2
- SFTZTYBXIXLRGQ-JBDRJPRFSA-N Ser-Ile-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O SFTZTYBXIXLRGQ-JBDRJPRFSA-N 0.000 description 2
- XNCUYZKGQOCOQH-YUMQZZPRSA-N Ser-Leu-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O XNCUYZKGQOCOQH-YUMQZZPRSA-N 0.000 description 2
- MUJQWSAWLLRJCE-KATARQTJSA-N Ser-Leu-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MUJQWSAWLLRJCE-KATARQTJSA-N 0.000 description 2
- GDUZTEQRAOXYJS-SRVKXCTJSA-N Ser-Phe-Asn Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CO)N GDUZTEQRAOXYJS-SRVKXCTJSA-N 0.000 description 2
- 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 2
- FZXOPYUEQGDGMS-ACZMJKKPSA-N Ser-Ser-Gln Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(O)=O FZXOPYUEQGDGMS-ACZMJKKPSA-N 0.000 description 2
- OZPDGESCTGGNAD-CIUDSAMLSA-N Ser-Ser-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CO OZPDGESCTGGNAD-CIUDSAMLSA-N 0.000 description 2
- KKKVOZNCLALMPV-XKBZYTNZSA-N Ser-Thr-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(O)=O KKKVOZNCLALMPV-XKBZYTNZSA-N 0.000 description 2
- BEBVVQPDSHHWQL-NRPADANISA-N Ser-Val-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O BEBVVQPDSHHWQL-NRPADANISA-N 0.000 description 2
- MQCPGOZXFSYJPS-KZVJFYERSA-N Thr-Ala-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O MQCPGOZXFSYJPS-KZVJFYERSA-N 0.000 description 2
- ZUXQFMVPAYGPFJ-JXUBOQSCSA-N Thr-Ala-Lys Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCCCN ZUXQFMVPAYGPFJ-JXUBOQSCSA-N 0.000 description 2
- TZKPNGDGUVREEB-FOHZUACHSA-N Thr-Asn-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O TZKPNGDGUVREEB-FOHZUACHSA-N 0.000 description 2
- MPUMPERGHHJGRP-WEDXCCLWSA-N Thr-Gly-Lys Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)O)N)O MPUMPERGHHJGRP-WEDXCCLWSA-N 0.000 description 2
- SPVHQURZJCUDQC-VOAKCMCISA-N Thr-Lys-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O SPVHQURZJCUDQC-VOAKCMCISA-N 0.000 description 2
- JMBRNXUOLJFURW-BEAPCOKYSA-N Thr-Phe-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N2CCC[C@@H]2C(=O)O)N)O JMBRNXUOLJFURW-BEAPCOKYSA-N 0.000 description 2
- KERCOYANYUPLHJ-XGEHTFHBSA-N Thr-Pro-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O KERCOYANYUPLHJ-XGEHTFHBSA-N 0.000 description 2
- IVDFVBVIVLJJHR-LKXGYXEUSA-N Thr-Ser-Asp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O IVDFVBVIVLJJHR-LKXGYXEUSA-N 0.000 description 2
- AHERARIZBPOMNU-KATARQTJSA-N Thr-Ser-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O AHERARIZBPOMNU-KATARQTJSA-N 0.000 description 2
- YRJOLUDFVAUXLI-GSSVUCPTSA-N Thr-Thr-Asp Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(O)=O YRJOLUDFVAUXLI-GSSVUCPTSA-N 0.000 description 2
- QJIODPFLAASXJC-JHYOHUSXSA-N Thr-Thr-Phe Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N)O QJIODPFLAASXJC-JHYOHUSXSA-N 0.000 description 2
- PELIQFPESHBTMA-WLTAIBSBSA-N Thr-Tyr-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CC1=CC=C(O)C=C1 PELIQFPESHBTMA-WLTAIBSBSA-N 0.000 description 2
- QGVBFDIREUUSHX-IFFSRLJSSA-N Thr-Val-Gln Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O QGVBFDIREUUSHX-IFFSRLJSSA-N 0.000 description 2
- MNYNCKZAEIAONY-XGEHTFHBSA-N Thr-Val-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O MNYNCKZAEIAONY-XGEHTFHBSA-N 0.000 description 2
- NKUIXQOJUAEIET-AQZXSJQPSA-N Trp-Asp-Thr Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@H](O)C)C(O)=O)=CNC2=C1 NKUIXQOJUAEIET-AQZXSJQPSA-N 0.000 description 2
- OBWQLWYNNZPWGX-QEJZJMRPSA-N Trp-Gln-Asp Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O OBWQLWYNNZPWGX-QEJZJMRPSA-N 0.000 description 2
- KDWZQYUTMJSYRJ-BHYGNILZSA-N Trp-Glu-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)N)C(=O)O KDWZQYUTMJSYRJ-BHYGNILZSA-N 0.000 description 2
- BEWOXKJJMBKRQL-AAEUAGOBSA-N Trp-Gly-Asp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)NCC(=O)N[C@@H](CC(=O)O)C(=O)O)N BEWOXKJJMBKRQL-AAEUAGOBSA-N 0.000 description 2
- WLQRIHCMPFHGKP-PMVMPFDFSA-N Trp-Leu-Phe Chemical compound C([C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CC=1C2=CC=CC=C2NC=1)CC(C)C)C(O)=O)C1=CC=CC=C1 WLQRIHCMPFHGKP-PMVMPFDFSA-N 0.000 description 2
- RCMHSGRBJCMFLR-BPUTZDHNSA-N Trp-Met-Asn Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(N)=O)C(O)=O)=CNC2=C1 RCMHSGRBJCMFLR-BPUTZDHNSA-N 0.000 description 2
- OOEUVMFKKZYSRX-LEWSCRJBSA-N Tyr-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N OOEUVMFKKZYSRX-LEWSCRJBSA-N 0.000 description 2
- PZXUIGWOEWWFQM-SRVKXCTJSA-N Tyr-Asn-Asn Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O PZXUIGWOEWWFQM-SRVKXCTJSA-N 0.000 description 2
- NSTPFWRAIDTNGH-BZSNNMDCSA-N Tyr-Asn-Tyr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O NSTPFWRAIDTNGH-BZSNNMDCSA-N 0.000 description 2
- JKUZFODWJGEQAP-KBPBESRZSA-N Tyr-Gly-Lys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)O)N)O JKUZFODWJGEQAP-KBPBESRZSA-N 0.000 description 2
- NOOMDULIORCDNF-IRXDYDNUSA-N Tyr-Gly-Phe Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O NOOMDULIORCDNF-IRXDYDNUSA-N 0.000 description 2
- GQVZBMROTPEPIF-SRVKXCTJSA-N Tyr-Ser-Asp Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O GQVZBMROTPEPIF-SRVKXCTJSA-N 0.000 description 2
- ZPFLBLFITJCBTP-QWRGUYRKSA-N Tyr-Ser-Gly Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(=O)NCC(O)=O ZPFLBLFITJCBTP-QWRGUYRKSA-N 0.000 description 2
- NAHUCETZGZZSEX-IHPCNDPISA-N Tyr-Trp-Asp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CC3=CC=C(C=C3)O)N NAHUCETZGZZSEX-IHPCNDPISA-N 0.000 description 2
- LVILBTSHPTWDGE-PMVMPFDFSA-N Tyr-Trp-Lys Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCCCN)C(O)=O)C1=CC=C(O)C=C1 LVILBTSHPTWDGE-PMVMPFDFSA-N 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- ASQFIHTXXMFENG-XPUUQOCRSA-N Val-Ala-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O ASQFIHTXXMFENG-XPUUQOCRSA-N 0.000 description 2
- LTFLDDDGWOVIHY-NAKRPEOUSA-N Val-Ala-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](C)NC(=O)[C@H](C(C)C)N LTFLDDDGWOVIHY-NAKRPEOUSA-N 0.000 description 2
- UUYCNAXCCDNULB-QXEWZRGKSA-N Val-Arg-Asn Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC(N)=O)C(O)=O UUYCNAXCCDNULB-QXEWZRGKSA-N 0.000 description 2
- ZMDCGGKHRKNWKD-LAEOZQHASA-N Val-Asn-Glu Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N ZMDCGGKHRKNWKD-LAEOZQHASA-N 0.000 description 2
- UDNYEPLJTRDMEJ-RCOVLWMOSA-N Val-Asn-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)NCC(=O)O)N UDNYEPLJTRDMEJ-RCOVLWMOSA-N 0.000 description 2
- UZDHNIJRRTUKKC-DLOVCJGASA-N Val-Gln-Val Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](C(C)C)C(=O)O)N UZDHNIJRRTUKKC-DLOVCJGASA-N 0.000 description 2
- 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 2
- DJEVQCWNMQOABE-RCOVLWMOSA-N Val-Gly-Asp Chemical compound CC(C)[C@@H](C(=O)NCC(=O)N[C@@H](CC(=O)O)C(=O)O)N DJEVQCWNMQOABE-RCOVLWMOSA-N 0.000 description 2
- LAYSXAOGWHKNED-XPUUQOCRSA-N Val-Gly-Ser Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O LAYSXAOGWHKNED-XPUUQOCRSA-N 0.000 description 2
- BVWPHWLFGRCECJ-JSGCOSHPSA-N Val-Gly-Tyr Chemical compound CC(C)[C@@H](C(=O)NCC(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N BVWPHWLFGRCECJ-JSGCOSHPSA-N 0.000 description 2
- VPGCVZRRBYOGCD-AVGNSLFASA-N Val-Lys-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O VPGCVZRRBYOGCD-AVGNSLFASA-N 0.000 description 2
- WSUWDIVCPOJFCX-TUAOUCFPSA-N Val-Met-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)N1CCC[C@@H]1C(=O)O)N WSUWDIVCPOJFCX-TUAOUCFPSA-N 0.000 description 2
- SSYBNWFXCFNRFN-GUBZILKMSA-N Val-Pro-Ser Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O SSYBNWFXCFNRFN-GUBZILKMSA-N 0.000 description 2
- VIKZGAUAKQZDOF-NRPADANISA-N Val-Ser-Glu Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O VIKZGAUAKQZDOF-NRPADANISA-N 0.000 description 2
- VHIZXDZMTDVFGX-DCAQKATOSA-N Val-Ser-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](C(C)C)N VHIZXDZMTDVFGX-DCAQKATOSA-N 0.000 description 2
- GBIUHAYJGWVNLN-UHFFFAOYSA-N Val-Ser-Pro Natural products CC(C)C(N)C(=O)NC(CO)C(=O)N1CCCC1C(O)=O GBIUHAYJGWVNLN-UHFFFAOYSA-N 0.000 description 2
- WUFHZIRMAZZWRS-OSUNSFLBSA-N Val-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](C(C)C)N WUFHZIRMAZZWRS-OSUNSFLBSA-N 0.000 description 2
- VVIZITNVZUAEMI-DLOVCJGASA-N Val-Val-Gln Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCC(N)=O VVIZITNVZUAEMI-DLOVCJGASA-N 0.000 description 2
- JVGDAEKKZKKZFO-RCWTZXSCSA-N Val-Val-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](C(C)C)N)O JVGDAEKKZKKZFO-RCWTZXSCSA-N 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 108010076324 alanyl-glycyl-glycine Proteins 0.000 description 2
- 108010069020 alanyl-prolyl-glycine Proteins 0.000 description 2
- 108010041407 alanylaspartic acid Proteins 0.000 description 2
- 108010005233 alanylglutamic acid Proteins 0.000 description 2
- 108010087924 alanylproline Proteins 0.000 description 2
- KOSRFJWDECSPRO-UHFFFAOYSA-N alpha-L-glutamyl-L-glutamic acid Natural products OC(=O)CCC(N)C(=O)NC(CCC(O)=O)C(O)=O KOSRFJWDECSPRO-UHFFFAOYSA-N 0.000 description 2
- 150000001413 amino acids Chemical group 0.000 description 2
- 108010008355 arginyl-glutamine Proteins 0.000 description 2
- 108010009111 arginyl-glycyl-glutamic acid Proteins 0.000 description 2
- 108010093581 aspartyl-proline Proteins 0.000 description 2
- 108010047857 aspartylglycine Proteins 0.000 description 2
- 108010068265 aspartyltyrosine Proteins 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 108010078144 glutaminyl-glycine Proteins 0.000 description 2
- XKUKSGPZAADMRA-UHFFFAOYSA-N glycyl-glycyl-glycine Natural products NCC(=O)NCC(=O)NCC(O)=O XKUKSGPZAADMRA-UHFFFAOYSA-N 0.000 description 2
- 108010033719 glycyl-histidyl-glycine Proteins 0.000 description 2
- 108010048994 glycyl-tyrosyl-alanine Proteins 0.000 description 2
- 108010050848 glycylleucine Proteins 0.000 description 2
- 108010037850 glycylvaline Proteins 0.000 description 2
- 108010040030 histidinoalanine Proteins 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 108010078274 isoleucylvaline Proteins 0.000 description 2
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 2
- 239000010977 jade Substances 0.000 description 2
- 108010044311 leucyl-glycyl-glycine Proteins 0.000 description 2
- 108010051673 leucyl-glycyl-phenylalanine Proteins 0.000 description 2
- 108010090333 leucyl-lysyl-proline Proteins 0.000 description 2
- 108010064235 lysylglycine Proteins 0.000 description 2
- 108010038320 lysylphenylalanine Proteins 0.000 description 2
- 108010017391 lysylvaline Proteins 0.000 description 2
- 108010056582 methionylglutamic acid Proteins 0.000 description 2
- 108010018625 phenylalanylarginine Proteins 0.000 description 2
- 230000009465 prokaryotic expression Effects 0.000 description 2
- 108010014614 prolyl-glycyl-proline Proteins 0.000 description 2
- 108010093296 prolyl-prolyl-alanine Proteins 0.000 description 2
- 108010031719 prolyl-serine Proteins 0.000 description 2
- 108010079317 prolyl-tyrosine Proteins 0.000 description 2
- 108010053725 prolylvaline Proteins 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 108091008146 restriction endonucleases Proteins 0.000 description 2
- 108010048397 seryl-lysyl-leucine Proteins 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 108010038745 tryptophylglycine Proteins 0.000 description 2
- 108010003137 tyrosyltyrosine Proteins 0.000 description 2
- 229920000936 Agarose Polymers 0.000 description 1
- CMLGVVWQQHUXOZ-GHCJXIJMSA-N Asn-Ala-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O CMLGVVWQQHUXOZ-GHCJXIJMSA-N 0.000 description 1
- PLOKOIJSGCISHE-BYULHYEWSA-N Asp-Val-Asn Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O PLOKOIJSGCISHE-BYULHYEWSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 229920001503 Glucan Polymers 0.000 description 1
- INCJJHQRZGQLFC-KBPBESRZSA-N Leu-Phe-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)NCC(O)=O INCJJHQRZGQLFC-KBPBESRZSA-N 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- BGWKULMLUIUPKY-BQBZGAKWSA-N Pro-Ser-Gly Chemical compound OC(=O)CNC(=O)[C@H](CO)NC(=O)[C@@H]1CCCN1 BGWKULMLUIUPKY-BQBZGAKWSA-N 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 108010028144 alpha-Glucosidases Proteins 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 101150030789 bglB gene Proteins 0.000 description 1
- 235000013877 carbamide Nutrition 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 239000012531 culture fluid Substances 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 1
- 239000007986 glycine-NaOH buffer Substances 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 229930027917 kanamycin Natural products 0.000 description 1
- 229960000318 kanamycin Drugs 0.000 description 1
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 1
- 229930182823 kanamycin A Natural products 0.000 description 1
- TWNIBLMWSKIRAT-VFUOTHLCSA-N levoglucosan Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@H]2CO[C@@H]1O2 TWNIBLMWSKIRAT-VFUOTHLCSA-N 0.000 description 1
- ZLNQQNXFFQJAID-UHFFFAOYSA-L magnesium carbonate Chemical compound [Mg+2].[O-]C([O-])=O ZLNQQNXFFQJAID-UHFFFAOYSA-L 0.000 description 1
- 239000001095 magnesium carbonate Substances 0.000 description 1
- 229910000021 magnesium carbonate Inorganic materials 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000012982 microporous membrane Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 239000006174 pH buffer Substances 0.000 description 1
- 239000005365 phosphate glass Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- XPFJYKARVSSRHE-UHFFFAOYSA-K trisodium;2-hydroxypropane-1,2,3-tricarboxylate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].[Na+].[Na+].OC(=O)CC(O)(C(O)=O)CC(O)=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O XPFJYKARVSSRHE-UHFFFAOYSA-K 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
- C12N9/2445—Beta-glucosidase (3.2.1.21)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/70—Vectors or expression systems specially adapted for E. coli
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
- C12P7/44—Polycarboxylic acids
- C12P7/46—Dicarboxylic acids having four or less carbon atoms, e.g. fumaric acid, maleic acid
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01021—Beta-glucosidase (3.2.1.21)
-
- 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
- C12N2800/00—Nucleic acids vectors
- C12N2800/10—Plasmid DNA
- C12N2800/101—Plasmid DNA for bacteria
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
本发明涉及一种β‑葡萄糖苷酶基因。所述β‑葡萄糖苷酶基因从土壤总DNA中获取，其核苷酸序列为SEQ ID NO：1。本发明的β‑葡萄糖苷酶不需要利用表面展示或者信号肽表达的技术就可直接使菌体利用纤维二糖，从而进行丁二酸的生产。
Description
技术领域
本发明涉及微生物技术领域，具体涉及一种β-葡萄糖苷酶基因及其应用。
背景技术
β-葡萄糖苷酶(EC3.2.1.21)属于水解酶。它可以水解并且结合末端、非还原性的β-D-葡萄糖及相应配基。该酶可以将纤维二糖或短链葡聚糖水解成葡萄糖。因其参与纤维素水解的最后一步，所以常被认为是纤维素水解糖化过程的限速酶。β-葡萄糖苷酶按照氨基酸序列可以划分为糖苷水解酶家族1和糖苷水解酶家族3两类。糖苷水解酶家族1中的酶主要来源于细菌、植物和哺乳动物；糖苷水解酶家族3中酶来源于真菌、细菌和植物。可见其来源比较广泛。β-葡萄糖苷酶一般通过表面展示和信号肽表达的方式达到在胞外分解纤维二糖为葡萄糖，以此供给细胞利用生长，目前尚未有β-葡萄糖苷酶不需要通过表面展示或者信号肽表达就可以利用纤维二糖生长的报道。
发明内容
本发明的技术目的之一，在于提供一种来自土壤总DNA的β-葡萄糖苷酶基因，所述的土壤总DNA采集自南京工业大学东操场边上小花园附近。
为实现上述技术目的，本发明采用如下技术方案：
本发明所述的β-葡萄糖苷酶基因，来源于南京工业大学东操场边上小花园附近土壤总DNA，其核苷酸序列如SEQ ID NO：1所示，其氨基酸序列如SEQ ID NO：2所示。
所述的β-葡萄糖苷酶基因是通过构建土壤总DNA的基因组文库，经过一系列酶切、酶连、转化等手段获得所需的阳性克隆，提取阳性克隆质粒测序，获得β-葡萄糖苷酶核苷酸序列。再根据所得到序列设计引物通过PCR即获得目的基因。优点是方便快捷效率高。
为获得本发明所述β-葡萄糖苷酶基因，具体步骤为：
(1)样品采集：土壤样品采自南京工业大学东操场边上小花园附近。
(2)土壤总DNA的提取：称取1g土壤样品，加入1mL 0.1mol/mL、pH 8.0磷酸缓冲液和玻璃珠，振荡1min，加入溶菌酶5mg，使溶菌酶最终浓度为2.5mg/mL，室温振荡15min，放置冰箱30min，加125μL 20％SDS振荡处理15min后离心，加酚(1:1体积)抽提1次，氯仿-异戊醇(1:1体积)抽提2次，加0.6体积异丙醇，室温放置1h，离心，70％乙醇清洗，30μLTE溶解。
(3)构建土壤总DNA的基因组文库：用限制性内切酶EcoRI将总DNA部分酶切，再经琼脂糖凝胶电泳分离回收2-3kb的DNA片段，与脱磷载体pUC118(BamHI/BAP)(TaKaRa，Code:D3321)连接后，化学转化导入制备好的E.coli DH5α感受态细胞，构建基因组文库。
限制性内切酶EcoRI酶切总DNA体系为：总DNA15μL，10×buffer H 3μL，EcoRI 3μL，加双蒸水调至总体积30μL。
37℃酶切30min。加入3μL 10×loading buffer终止酶切反应。酶切产物经1.5％琼脂糖凝胶电泳进行分离，回收2-3kb DNA片段。回收按照试剂盒说明书进行。
酶切片段与脱磷载体pUC118(BamHI/BAP)的连接：
将1μL pUC118(BamHI/BAP)载体DNA转移到无菌微量离心管中，加入6μL总DNA的EcoRI酶切回收片段，加水至8.5μL，于45℃加温5min使重新退火的粘端解链，将混合物冷却到0℃。然后加入1μL 10×T4DNA连接酶缓冲液，0.5μL T4DNA连接酶。16℃连接反应12h以上。
(4)酶连产物的转化和获取阳性克隆：将10μL酶连产物加入到200μL在冰上融化后的E.coli DH5α感受态细胞中，冰浴30min，在42℃水浴锅中热激90s后，快速转移到冰浴中冷却1～2min，向每管中加入800μL液体LB培养基，37℃摇床80-90rpm温育45min，复苏细胞。4000rpm离心3min，剩余200μL感受态细胞涂布于含8mM pNPG和100mg/l氨苄青霉素的LB琼脂平板上，平板倒置于37℃培养箱培养，12-16h后出现菌落，选择有天然变色的克隆子，即为含β-葡萄糖苷酶基因片段的阳性克隆子，同时对阳性克隆子进行编号。挑单菌落接种于5mL LB液体试管，待长出菌悬液后提质粒验证。
(5)提取阳性克隆质粒测序，获取β-葡萄糖苷酶核苷酸序列：阳性克隆子***片段的测序委托南京金斯瑞生物科技有限公司完成。测序引物为pUC118(BamHI/BAP)载体上特异性引物，并根据两端测序结果设计引物，直至将整个***片段测通。
将测序完成的序列在NCBI数据库中进行比对，并利用ORF finder工具鉴定***片段所含的ORF。
(6)引物的设计与合成：根据测序结果的基因序列设计PCR引物，引物设计如下：
设计长度为50mer左右的引物，该引物分为两个部分，P1的第一部分是上游载体末端同源序列，第二部分是基因特异性正向扩增序列。P2的第一部分是基因特异性反向扩增序列，第二部分是下游载体末端同源序列。
利用PCR进行β-葡萄糖苷酶基因扩增，在50μL反应体系中，P1、P2两个引物的添加量为1μL，扩增条件为：94℃预热10min；94℃，45s；60℃，45s；72℃，2.5min；72℃，10min，中间三步共30个循环，使用的DNA聚合酶为2×Phanta@Master Mix酶(诺唯赞公司，中国)。
PCR结束后，1.5％琼脂糖胶回收片段，测定浓度。同时对于表达载体pET-28a(+)用限制性内切酶EcoRI酶切线性化，1.5％琼脂糖胶回收片段，测定浓度。计算最适克隆载体使用量(＝0.02×克隆载体碱基对数)ng和最适***片段使用量(＝0.04×***片段碱基对数)ng(例如，将长度为2kb的***片段克隆至长度为5kb的克隆载体时，克隆载体的最适使用量应为：0.02×5000＝100ng；***片段最适使用量应为：0.04×2000＝80ng)。利用One Step Cloning Kit试剂盒(诺唯赞公司)进行重组反应。37℃反应30min后转化到DH5α感受态中。获得长度为2775bp的SEQ ID NO：1序列的阳性克隆，即为本发明的β-葡萄糖苷酶基因。
本发明还提供了一种重组载体，所述重组载体含有上述的β-葡萄糖苷酶基因。
本发明还提供了一种宿主细胞，所述宿主细胞含有上述重组载体。
所述宿主细胞可为原核细胞。
所述原核细胞可为大肠杆菌E.coli BL21(DE3)。
为实现上述技术目的，具体可采用如下技术方案：将上述SEQ ID NO：1序列直接与原核表达载体pET-28a(+)连接，37℃反应30min。将该载体转化感受态大肠杆菌DH5α。将菌液涂布在含30μg/mL卡那霉素霉素的固体LB培养基(2％蛋白胨，1％酵母粉，1％NaCl，1～2％琼脂)培养后获得单菌落。挑取单菌落用5mL液体LB试管培养过夜，提质粒，进行单双酶切验证。选取验证正确质粒转化到大肠杆菌E.coli BL21(DE3)。将菌液涂布含30μg/mL卡那霉素的固体LB培养基培养后获得单菌落。
本发明还提供了本发明所述β-葡萄糖苷酶基因在生产丁二酸中的应用。
本发明的β-葡萄糖苷酶是一种新型β-葡萄糖苷酶，它是一类应用非常广泛的生物酶类。通过对该基因的原核表达和纯化，可以应用于该酶的工业化生产中。本发明的β-葡萄糖苷酶不需要利用表面展示或者信号肽表达的技术就可直接使菌体利用纤维二糖，从而进行丁二酸的生产。
附图说明
图1为以本发明β-葡萄糖苷酶基因在E.coli BL21(pET-28a(+))中高效表达实验方案图。
图2为本发明β-葡萄糖苷酶的最适温度曲线图。
图3位本发明β-葡萄糖苷酶的温度的稳定性曲线图。
图4为本发明β-葡萄糖苷酶的最适pH曲线图。
图5为本发明β-葡萄糖苷酶的pH的稳定性曲线图。
图6为金属离子对本发明β-葡萄糖苷酶酶活的影响柱状图。
图7为有机溶剂对本发明β-葡萄糖苷酶酶活的影响柱状图。
具体实施方式
以下结合附图详细描述本发明的技术方案。实施例仅用以说明本发明的技术方案而非限制，尽管参照较佳实施例对本发明进行详细说明，本领域的普通技术人员应当理解，可以对发明的技术方案进行修改或者等同替换，而不脱离本发明技术方案的精神和范围，其均应涵盖在本发明的权利要求范围中。
本发明所用的试剂若未经说明，均购自西格玛-奥奥德里奇(Sigma-Aldrich)公司。
本发明涉及分子生物实验，若未特别注明，均参考自《分子克隆实验指南》一书(J.萨姆布鲁克、D.W.拉塞尔著，2002，科学出版社。)
实施例1获取β-葡萄糖苷酶基因
本实施例说明获取本发明所述β-葡萄糖苷酶基因的方法。具体步骤如下：
(1)样品采集：土壤样品采自南京工业大学东操场边上小花园附近。
(2)土壤总DNA的提取：称取1g土壤样品，加入1mL 0.1mol/mL、pH 8.0磷酸缓冲液和玻璃珠，振荡1min，加入溶菌酶5mg，使溶菌酶最终浓度为2.5mg/mL，室温振荡15min，放置冰箱30min，加125μL 20％SDS振荡处理15min后离心，加酚(1∶1体积)抽提1次，氯仿-异戊醇(1∶1体积)抽提2次，加0.6体积异丙醇，室温放置1h，离心，70％乙醇清洗，30μLTE溶解。
(3)构建土壤总DNA的基因组文库：用限制性内切酶EcoRI将总DNA部分酶切，再经琼脂糖凝胶电泳分离回收2-3kb的DNA片段，与脱磷载体pUC118(BamHI/BAP)(TaKaRa，Code:D3321)连接后，化学转化导入制备好的E.coli DH5α感受态细胞，构建基因组文库。
限制性内切酶EcoRI酶切总DNA体系为：总DNA15μL，10×buffer H 3μL，EcoRI 3μL，加双蒸水调至总体积30μL。
37℃酶切30min。加入3μL 10×loading buffer终止酶切反应。酶切产物经1.5％琼脂糖凝胶电泳进行分离，回收2-3kb DNA片段。回收按照试剂盒说明书进行。
酶切片段与脱磷载体pUC118(BamHI/BAP)的连接：
将1μL pUC118(BamHI/BAP)载体DNA转移到无菌微量离心管中，加入6μL总DNA的ECORI酶切回收片段，加水至8.5μL，于45℃加温5min使重新退火的粘端解链，将混合物冷却到0℃。然后加入1μL 10×T4DNA连接酶缓冲液，0.5μL T4DNA连接酶。16℃连接反应12h以上。
(4)酶连产物的转化和获取阳性克隆：将10μL酶连产物加入到200μL在冰上融化后的E.coli DH5α感受态细胞中，冰浴30min，在42℃水浴锅中热激90s后。快速转移到冰浴中冷却1～2min，向每管中加入800μL液体LB培养基，37℃摇床80-90rpm温育45min，复苏细胞。4000rpm离心3min，剩余200μL感受态细胞涂布于含8mM pNPG和100mg/l氨苄青霉素的LB琼脂平板上，平板倒置于37℃培养箱培养，12-16h后出现菌落，选择有天然变色的克隆子，即为含β-葡萄糖苷酶基因片段的阳性克隆子，同时对阳性克隆子进行编号。挑单菌落接种于5mL LB液体试管，待长出菌悬液后提质粒验证。
(5)提取阳性克隆质粒测序，获取β-葡萄糖苷酶核苷酸序列：阳性克隆子***片段的测序委托南京金斯瑞生物科技有限公司完成。测序引物为pUC118(BamHI/BAP)载体上特异性引物，并根据两端测序结果设计引物，直至将整个***片段测通。
将测序完成的序列在NCBI数据库中进行比对，并利用ORF finder工具鉴定***片段所含的ORF。
(6)引物的设计与合成：根据测序结果的基因序列设计PCR引物，利用PCR进行β-葡萄糖苷酶基因扩增，PCR结束后，用1.5％琼脂糖胶回收片段，测定浓度，即获取本发明所述β-葡萄糖苷酶基因。
实施例2β-葡萄糖苷酶基因的扩增
本实施例说明β-葡萄糖苷酶基因扩增的具体过程。
参看图1，用PCR方式来克隆本发明的β-葡萄糖苷酶基因。设计的引物分别如下：
上游引物P1：Tm＝66.7，45mer；引物序列如下：
CAAATGGGTCGCGGATCCGAATTCATGAAAGTAAAATCAACATGG；
下游引物P2:Tm＝68.7，52mer；引物序列如下：
TTGTCGACGGAGCTCGAATTCTTACTTTTTTGCCTTTTCTGTAGAGGTTGCC；
利用PCR进行β-葡萄糖苷酶基因扩增，在50μL反应体系中，P1、P2两个引物的添加量为1μL，扩增条件为：94℃预热10min；94℃，45s；55℃，45s；72℃，2.5min；72℃，10min，中间三步共30个循环，使用的DNA聚合酶为2×Phanta@Master Mix酶(诺唯赞公司，中国)。
PCR结束后，1.5％琼脂糖胶回收片段，测定浓度。计算最适克隆载体使用量(＝0.02×克隆载体碱基对数)ng和最适***片段使用量(＝0.04×***片段碱基对数)ng，利用One StepCloning Kit试剂盒(诺唯赞公司)进行重组反应。37℃反应30min后转化到DH5α感受态中。获得长度为2775bp的阳性克隆，即为本发明的β-葡萄糖苷酶基因，其核苷酸序列如SEQ IDNO：1所示。
实施例3β-葡萄糖苷酶基因在大肠杆菌中表达
将上述SEQ ID NO：1序列直接与原核表达载体pET-28a(+)连接，37℃反应30min。将该载体转化感受态DH5α。将菌液涂布含30μg/mL氨苄青霉素的固体LB培养基(质量分数配方为：2％蛋白胨，1％酵母粉，1％NaCl，1～2％琼脂)培养后获得单菌落。挑取单菌落用5mL液体LB试管培养过夜，提质粒，进行单双酶切验证。选取验证正确质粒转化到大肠杆菌E.coliBL21(DE3)。将菌液涂布含30μg/mL氨苄青霉素的固体LB培养基培养后获得单菌落。再挑取单菌落用5mL液体LB试管培养过夜，提质粒，进行单双酶切验证，选取验证正确的菌落，接种到50mL液体LB培养基(2％蛋白胨，1％酵母粉，1％NaCl)中，37℃摇床培养，直到菌液浓度达到OD600为0.6～0.8时，用终浓度为0.3mM的异丙基-β-D-硫代半乳糖苷(IPTG)进行诱导表达，25℃，150rpm，诱导20h。
实施例4β-葡萄糖苷酶的纯化
将实施例3最终获得的培养液在4℃，8000rpm离心10min获得菌体，用预冷的50mMTris-HCl缓冲液(PH＝8.5)洗涤三次，最后用预冷的100mMTris-HCl缓冲液(PH＝8.5)重悬。超声破碎得到初酶液，4℃下，12000rpm离心30min，用微孔滤膜过滤离心好的上清，用Ni-Agarose柱子纯化出目的蛋白。将目的蛋白透析后用蛋白保存液保存起来。
实施例5β-葡萄糖苷酶的酶动力学特性分析
β-葡萄糖苷酶动力学特性分析主要底物为pNPG。反应采用实施例4中的体系。主要测定酶对底物的Km和最大反应速度Vmax。也就是说在温度、pH及酶浓度恒定的条件下，底物浓度对酶促反应的速度有很大的影响。在底物浓度很低时，酶促反应的速度(v)随底物浓度的增加而迅速增加；随着底物浓度的继续增加，反应速度的增加开始减慢；当底物浓度增加到某种程度时，反应速度达到一个极限值(Vmax)。
底物浓度与反应速度的这种关系可用Michaelis-Menten方程式表示：
式中：v-反应速度；Km-米氏常数；Vmax-酶反应最大速度；[S]-底物浓度。
采用Linewaver-Burk作图法测定Km、Vmax，该法是根据米氏方程的倒数形式，以1/v对1/[S]作图，得到一条直线。直线在横轴上的截距为-1/Km，纵截距为1/Vmax，求出Km与Vmax。所述方程如下所示：
在这里我们所用的底物的浓度是：pNPG(从0.5mM到8mM)。所得到的结果如表1所示。
表1
实施例6β-葡萄糖苷酶的生理生化特性分析
对实施例4中保存的蛋白酶液做特定的生理生化特性分析。本过程是在下述酶反应体系中进行的：80μL的100mM Tris-HCl缓冲液(pH＝8.5)加入100μL的8mM的pNPG底物，40℃预热2min后，加入20μL适当稀释的酶液开始反应，在40℃下反应10min，加入100μL的0.5M的Na2CO3终止反应。然后在405nm处测量对硝基酚的释放量。在这里我们定义一定数量的酶在每分钟催化1μM的对硝基酚的活力为1U。β-葡萄糖苷酶的蛋白浓度是使用Micro-Spectrophotometer K5500来分析的。
我们分析的生理生化特性主要有酶的最适温度，酶的温度稳定性，酶的最适pH，pH的稳定性，金属离子对酶活的影响，有机溶剂对酶活的影响。图中相对活性是指在最适温度和最适pH条件下测定得到的酶活，以此为100％的基准，其他条件下测得活性与之比较从而得到的比值即为该条件下的相对活性。
在研究酶的最适反应的温度时，反应的温度从20℃开始，每隔10℃，一直到80℃。每个体系做3个重复试验(以下每个反应都是3次重复)，研究结果如图2所示，从图中可知，本发明的β-葡萄糖苷酶在20-50℃具有较高酶活，其中最适温度为40℃，温度超过50℃的时候酶活力快速下降，超过60℃后基本没有酶活。
在研究酶的温度稳定性时，主要测定酶在20，30，40和50℃这四个不同温度处理酶液，然后在最适温度下测定酶的残余活性，研究结果如图3所示，从图中可知，本发明的β-葡萄糖苷酶酶活力随着时间的增长逐渐降低。在20℃条件下酶活最稳定，30-40℃条件下活力依次下降，在50℃的情况下，酶稳定性迅速降低至无活性。可见较低的温度更适合保存该酶。
在研究酶的最适pH时，选择不同的缓冲液，主要有柠檬酸-柠檬酸三钠缓冲液(0.1M，pH4.0-6.0)，磷酸盐缓冲液(0.1M，pH6.0-8.0)，Tris-HCl缓冲液(0.05M，pH8.0-9.0)，甘氨酸-NaOH缓冲液(0.05M，pH8.5-10.0)；研究结果如图4所示，从图中可知，不同pH对酶活力有影响，当pH处于酸性环境下，酶活力较低。其中最适pH为8.5，该酶在碱性环境下比在酸性环境下有更好的酶活力。
在研究酶的pH稳定性时，是先将酶在不同的pH缓冲液中，在4℃下处理24h，然后在最适pH下测定酶的残余活性；研究结果如图5所示，从图中可知，该酶在碱性条件下比酸性条件下更稳定，且在最适pH下稳定性最好。
在金属离子对酶活的影响，有机溶剂对酶活的影响是将酶用不同的影响因素处理，然后在酶的最佳反应体系中测定酶的残余活性。研究结果如图6-图7所示，从图中可以看出，基本所有的金属离子对该酶都有激活作用，其中Fe2+的激活作用最明显；有机溶剂中除了SDS和Tritonx-100对该酶有激活作用，其他有机试剂都起到抑制作用，尿素的抑制作用最强。
经实验验证，酶在40℃，pH8.5时活性最高。
实施例7
本实施例通过实验描述β-葡萄糖苷酶基因在生产丁二酸中的应用，来说明本发明β-葡萄糖苷酶应用于工业化生产的优越性。
采用本发明β-葡萄糖苷酶基因生产丁二酸的步骤具体如下：
(1)构建质粒pTrc99a-bglB，导入大肠杆菌DH5α感受态中。
其中β-葡萄糖苷酶基因(bglB)的获取方法。具体步骤如下：
①样品采集：土壤样品采自南京工业大学东操场边上小花园附近。
②土壤总DNA的提取：称取1g土壤样品，加入1mL 0.1mol/mL、pH 8.0磷酸缓冲液和玻璃珠，振荡1min，加入溶菌酶5mg，使溶菌酶最终浓度为2.5mg/mL，室温振荡15min，放置冰箱30min，加125μL 20％SDS振荡处理15min后离心，加酚(1∶1体积)抽提1次，氯仿-异戊醇(1∶1体积)抽提2次，加0.6体积异丙醇，室温放置1h，离心，70％乙醇清洗，30μLTE溶解。
③构建土壤总DNA的基因组文库：用限制性内切酶EcoRI将总DNA部分酶切，再经琼脂糖凝胶电泳分离回收2-3kb的DNA片段，与脱磷载体pUC118(BamHI/BAP)(TaKaRa，Code:D3321)连接后，化学转化导入制备好的E.coli DH5α感受态细胞，构建基因组文库。
限制性内切酶EcoRI酶切总DNA体系为：总DNA15μL，10×buffer H 3μL，EcoRI 3μL，加双蒸水调至总体积30μL。
37℃酶切30min。加入3μL 10×loading buffer终止酶切反应。酶切产物经1.5％琼脂糖凝胶电泳进行分离，回收2-3kb DNA片段。回收按照试剂盒说明书进行。
酶切片段与脱磷载体pUC118(BamHI/BAP)的连接：
将1μL pUC118(BamHI/BAP)载体DNA转移到无菌微量离心管中，加入6μL总DNA的ECORI酶切回收片段，加水至8.5μL，于45℃加温5min使重新退火的粘端解链，将混合物冷却到0℃。然后加入1μL 10×T4DNA连接酶缓冲液，0.5μL T4DNA连接酶。16℃连接反应12h以上。
④酶连产物的转化和获取阳性克隆：将10μL酶连产物加入到200μL在冰上融化后的E.coli DH5α感受态细胞中，冰浴30min，在42℃水浴锅中热激90s后。快速转移到冰浴中冷却1～2min，向每管中加入800μL液体LB培养基，37℃摇床80-90rpm温育45min，复苏细胞。4000rpm离心3min，剩余200μL感受态细胞涂布于含8mM pNPG和100mg/l氨苄青霉素的LB琼脂平板上，平板倒置于37℃培养箱培养，12-16h后出现菌落，选择有天然变色的克隆子，即为含β-葡萄糖苷酶基因片段的阳性克隆子，同时对阳性克隆子进行编号。挑单菌落接种于5mL LB液体试管，待长出菌悬液后提质粒验证。
⑤提取阳性克隆质粒测序，获取β-葡萄糖苷酶核苷酸序列：阳性克隆子***片段的测序委托南京金斯瑞生物科技有限公司完成。测序引物为pUC118(BamHI/BAP)载体上特异性引物，并根据两端测序结果设计引物，直至将整个***片段测通。
将测序完成的序列在NCBI数据库中进行比对，并利用ORF finder工具鉴定***片段所含的ORF。
⑥用PCR方式来克隆本发明的β-葡萄糖苷酶基因。设计的引物分别如下：
上游引物P1：Tm＝66，40mer；引物序列如下：
GCCTGCAGGTCGACTCTAGATTACTTTTTTGCCTTTTCTG；
下游引物P2:Tm＝63，40mer；引物序列如下：
GGAAACAGACCATGGAATTCGTGAAAGTAAAATCAACATG；
利用PCR进行β-葡萄糖苷酶基因扩增，在50μL反应体系中，P1、P2两个引物的添加量为1μL，扩增条件为：94℃预热10min；94℃，45s；55℃，45s；72℃，2.5min；72℃，10min，中间三步共30个循环，使用的DNA聚合酶为2×Phanta@Master Mix酶(诺唯赞公司，中国)。
PCR结束后，1.5％琼脂糖胶回收片段，测定浓度。同时对于表达载体pTrc99a用限制性内切酶EcoRI和XbaI酶切线性化，1.5％琼脂糖胶回收片段，测定浓度。计算最适克隆载体使用量(＝0.02×克隆载体碱基对数)ng和最适***片段使用量(＝0.04×***片段碱基对数)ng，利用One Step Cloning Kit试剂盒(诺唯赞公司)进行重组反应。37℃反应30min后转化到DH5α感受态中。
(2)提取质粒，验证质粒正确后，将正确的质粒导入大肠杆菌suc260感受态中，继续验证，完成菌株构建suc260-pTrc99a-bglB。
(3)以菌suc260-pTrc99a为对照菌，进行厌氧发酵。先在5mL LB试管里过夜生长，再转接到50mL LB的摇瓶中，37℃，200rpm生长，至OD600为0.6～0.8时用IPTG诱导，终浓度为0.3mM；25℃，150rpm培养6～8h。用离心管收集菌液后用纯水洗3遍，目的是为了洗掉原来的LB培养基。最后用纯水重悬菌泥，使最终OD550控制在4左右，然后向血清瓶中加入菌液，接种量为10％，其中，血清瓶内的培养基为AM1培养基，唯一碳源为30g/L的纤维二糖，装液量为30mL。接种前用CO2气体预先混匀培养基中的纤维二糖和碱式碳酸镁，约混匀1min,然后进行接种，取初样后向每个血清瓶中再次通CO2气体，目的是是菌液和培养基充分混匀并且保证厌氧状态。37℃，180rpm培养。培养结束后取终样。经检测，发酵产物中丁二酸的质量收率为0.85g/g(即丁二酸对纤维二糖的收率)。
序列表
<110> 南京工业大学
<120> 一种β-葡萄糖苷酶基因及其应用
<130> xb16061401
<160> 2
<170> PatentIn version 3.5
<210> 1
<211> 2730
<212> DNA
<213> Artificial Sequence
<220>
<223> 1
<220>
<221> CDS
<222> (1)..(2730)
<400> 1
gtg aaa gta aaa tca aca tgg ttg ttc cgt atg gtg atg atg aca gtc 48
Val Lys Val Lys Ser Thr Trp Leu Phe Arg Met Val Met Met Thr Val
1 5 10 15
att gcg gct gtt gtg atc gga cct atg cat att gca gga gcg gca gga 96
Ile Ala Ala Val Val Ile Gly Pro Met His Ile Ala Gly Ala Ala Gly
20 25 30
agc cgg aca gat cgg cct tgg atg aac acg tcc ctg tcg gct gag aaa 144
Ser Arg Thr Asp Arg Pro Trp Met Asn Thr Ser Leu Ser Ala Glu Lys
35 40 45
cgg aca gca ttg ctg tta caa gaa atg acg ctg gaa gaa aaa ata gat 192
Arg Thr Ala Leu Leu Leu Gln Glu Met Thr Leu Glu Glu Lys Ile Asp
50 55 60
ctg gtg act ggt aag gtc aac aat tat tat gga ttt tat aat aat tcg 240
Leu Val Thr Gly Lys Val Asn Asn Tyr Tyr Gly Phe Tyr Asn Asn Ser
65 70 75 80
atg gag cgg ttg ggt att cca gcg tta aag atg gca gat gga cct gcg 288
Met Glu Arg Leu Gly Ile Pro Ala Leu Lys Met Ala Asp Gly Pro Ala
85 90 95
ggt gtg cgg atc gcc aat ccg gat gtg cag gac aag caa tca act gcg 336
Gly Val Arg Ile Ala Asn Pro Asp Val Gln Asp Lys Gln Ser Thr Ala
100 105 110
ttg ccg gcc cct att gca ctg gcc gca acc tgg gat acc caa gct gcc 384
Leu Pro Ala Pro Ile Ala Leu Ala Ala Thr Trp Asp Thr Gln Ala Ala
115 120 125
aaa cag tat gga gat ttg ctt gga gac gag gct ttt aat aca act cat 432
Lys Gln Tyr Gly Asp Leu Leu Gly Asp Glu Ala Phe Asn Thr Thr His
130 135 140
aat gtg gtg ctt gga ccg ggg atg gat att gcc cgt att cca tgg gga 480
Asn Val Val Leu Gly Pro Gly Met Asp Ile Ala Arg Ile Pro Trp Gly
145 150 155 160
tcg agg aac ttt gaa tct atg ggc gag gac ccg cta ctg caa tcg caa 528
Ser Arg Asn Phe Glu Ser Met Gly Glu Asp Pro Leu Leu Gln Ser Gln
165 170 175
atg gcg acc gct tat gtc aaa ggt gta caa agc cac cct gtc ttg gcg 576
Met Ala Thr Ala Tyr Val Lys Gly Val Gln Ser His Pro Val Leu Ala
180 185 190
act gcg aaa cat tac ctt atg aat aat cag gag acg gag cgt ttc acg 624
Thr Ala Lys His Tyr Leu Met Asn Asn Gln Glu Thr Glu Arg Phe Thr
195 200 205
aca aat gtc aaa gtc agt gac cgt gcg ctg cat gaa att tac ata cgc 672
Thr Asn Val Lys Val Ser Asp Arg Ala Leu His Glu Ile Tyr Ile Arg
210 215 220
ccg ttt gag gag gct att gcc aag gca gat ttg ggc gga gct atg tgc 720
Pro Phe Glu Glu Ala Ile Ala Lys Ala Asp Leu Gly Gly Ala Met Cys
225 230 235 240
tcc ttt aac aag gtc aac ggc gaa tcg gca tgt gag aac aag acc atc 768
Ser Phe Asn Lys Val Asn Gly Glu Ser Ala Cys Glu Asn Lys Thr Ile
245 250 255
ctg aca gac ctt ttg aaa aag gaa atg caa ttc aag ggt ttc gtg atg 816
Leu Thr Asp Leu Leu Lys Lys Glu Met Gln Phe Lys Gly Phe Val Met
260 265 270
agt gat tat ggc gcg aat ctg agt aca gtc gaa tcg gca aat agc ggc 864
Ser Asp Tyr Gly Ala Asn Leu Ser Thr Val Glu Ser Ala Asn Ser Gly
275 280 285
ttg gac ctg gaa aca ccg gga acc cct tat ggt aaa tgg gga gat caa 912
Leu Asp Leu Glu Thr Pro Gly Thr Pro Tyr Gly Lys Trp Gly Asp Gln
290 295 300
ctg ctg gat gcg gtc aag act ggt aaa gta agt gaa caa acg att gat 960
Leu Leu Asp Ala Val Lys Thr Gly Lys Val Ser Glu Gln Thr Ile Asp
305 310 315 320
gat aag gct aag cgt att ctg gtg cag atg ttc agt aaa ggt ttg ttt 1008
Asp Lys Ala Lys Arg Ile Leu Val Gln Met Phe Ser Lys Gly Leu Phe
325 330 335
gat cac ccg gct caa aat aat caa att gat gcc cgt gac cat ggt aaa 1056
Asp His Pro Ala Gln Asn Asn Gln Ile Asp Ala Arg Asp His Gly Lys
340 345 350
aca gcc cgt caa ttg gct gag gaa agt atg gtg ctg ctt caa aat aaa 1104
Thr Ala Arg Gln Leu Ala Glu Glu Ser Met Val Leu Leu Gln Asn Lys
355 360 365
aac aac gta tta ccg ttg tct caa gat aaa ctc aaa tcc att gct gta 1152
Asn Asn Val Leu Pro Leu Ser Gln Asp Lys Leu Lys Ser Ile Ala Val
370 375 380
att ggt cca gat gca gat aac ggc acc gtg gca ggc gga ggc agt tcg 1200
Ile Gly Pro Asp Ala Asp Asn Gly Thr Val Ala Gly Gly Gly Ser Ser
385 390 395 400
ttg gtg aac ccg acg tac acg gtg agt ccg ctg gag ggc att cgt aat 1248
Leu Val Asn Pro Thr Tyr Thr Val Ser Pro Leu Glu Gly Ile Arg Asn
405 410 415
cgt gta ggc aaa gga gtt acc gtc cag tat gca ccg ggc acg gat cct 1296
Arg Val Gly Lys Gly Val Thr Val Gln Tyr Ala Pro Gly Thr Asp Pro
420 425 430
att tct gcg ggg gac att atg cca ggt cca tca gct gtt cca tct tcg 1344
Ile Ser Ala Gly Asp Ile Met Pro Gly Pro Ser Ala Val Pro Ser Ser
435 440 445
ttg ctg acc aca tcc gat cag aag gag aat atc agt gtt gga tat gcg 1392
Leu Leu Thr Thr Ser Asp Gln Lys Glu Asn Ile Ser Val Gly Tyr Ala
450 455 460
act tat ggc gat gct gag cag gga ctg cga ggc gaa tac tgg aag aat 1440
Thr Tyr Gly Asp Ala Glu Gln Gly Leu Arg Gly Glu Tyr Trp Lys Asn
465 470 475 480
aac aag atg gaa ggc aat ccg att ctt gtg cgc aat gat gat caa gtc 1488
Asn Lys Met Glu Gly Asn Pro Ile Leu Val Arg Asn Asp Asp Gln Val
485 490 495
aat atg aac ctt ggc ttt tat aat tat cag ggc ttc aat gcg aag tcc 1536
Asn Met Asn Leu Gly Phe Tyr Asn Tyr Gln Gly Phe Asn Ala Lys Ser
500 505 510
cct aaa gtt cct aat aca ccc acg acc ctg aac ggt ttg ata tct gca 1584
Pro Lys Val Pro Asn Thr Pro Thr Thr Leu Asn Gly Leu Ile Ser Ala
515 520 525
cgc tgg aca ggg gct att gca gct ccc aag gat ggt gac tat gca ttg 1632
Arg Trp Thr Gly Ala Ile Ala Ala Pro Lys Asp Gly Asp Tyr Ala Leu
530 535 540
tca tta acc agt ttg gga tca agc aag ctg tat ctt gat gat aag cta 1680
Ser Leu Thr Ser Leu Gly Ser Ser Lys Leu Tyr Leu Asp Asp Lys Leu
545 550 555 560
ttt gta gat aat cag ggt acg aag ctg gaa aca acg aag aaa aac atc 1728
Phe Val Asp Asn Gln Gly Thr Lys Leu Glu Thr Thr Lys Lys Asn Ile
565 570 575
tcg ttc aaa gcg ggg gaa aaa cat aag ata cgt att gaa tat cgt gcc 1776
Ser Phe Lys Ala Gly Glu Lys His Lys Ile Arg Ile Glu Tyr Arg Ala
580 585 590
gat tat cca aca aat ggt cgc gat tct ggc ggt atg gtt cgt ctg ggc 1824
Asp Tyr Pro Thr Asn Gly Arg Asp Ser Gly Gly Met Val Arg Leu Gly
595 600 605
tgg gag cct ccg gca gat acg aca gac aag ctg att gac aat gct gta 1872
Trp Glu Pro Pro Ala Asp Thr Thr Asp Lys Leu Ile Asp Asn Ala Val
610 615 620
aag ctg gcc aaa aaa tcg gat gtt gcg att gtg gtc acg cgt acg tat 1920
Lys Leu Ala Lys Lys Ser Asp Val Ala Ile Val Val Thr Arg Thr Tyr
625 630 635 640
gag agt gaa ggc tat gtg gag cgt tcc gat atg gag ctg cca aac aat 1968
Glu Ser Glu Gly Tyr Val Glu Arg Ser Asp Met Glu Leu Pro Asn Asn
645 650 655
cag gat cgg ctg atc cgt gcg gtt gcc gct gcc aat ccg aaa acc att 2016
Gln Asp Arg Leu Ile Arg Ala Val Ala Ala Ala Asn Pro Lys Thr Ile
660 665 670
gtg gta caa atg agc ggc aga gct gtg caa atg gac acc tgg cag gac 2064
Val Val Gln Met Ser Gly Arg Ala Val Gln Met Asp Thr Trp Gln Asp
675 680 685
aaa gtt cct gcg atc gta cag gca tgg ttt gct ggt caa gag caa ggg 2112
Lys Val Pro Ala Ile Val Gln Ala Trp Phe Ala Gly Gln Glu Gln Gly
690 695 700
aat gcc ata gcg cgc gtc ttg ttt ggg gat gta aat cct tct ggc aag 2160
Asn Ala Ile Ala Arg Val Leu Phe Gly Asp Val Asn Pro Ser Gly Lys
705 710 715 720
cta ccg gta acc ttc ccg gtt aac gag cag tct aca cct gta tcc tca 2208
Leu Pro Val Thr Phe Pro Val Asn Glu Gln Ser Thr Pro Val Ser Ser
725 730 735
ccg gag aaa ttt ccg ggt gtc aat gga gtg ggt gac tat tca gat ggt 2256
Pro Glu Lys Phe Pro Gly Val Asn Gly Val Gly Asp Tyr Ser Asp Gly
740 745 750
att ttt gta ggc tac cgt gga tat gaa aaa tcg ggt atc caa cct gcg 2304
Ile Phe Val Gly Tyr Arg Gly Tyr Glu Lys Ser Gly Ile Gln Pro Ala
755 760 765
ttc tcc ttt gga cac ggt tta tcc tac acg act ttc gga tac agt gat 2352
Phe Ser Phe Gly His Gly Leu Ser Tyr Thr Thr Phe Gly Tyr Ser Asp
770 775 780
ttg aag gta aaa cag cat gcg agt gga aag aaa tcc gat cgt aca agt 2400
Leu Lys Val Lys Gln His Ala Ser Gly Lys Lys Ser Asp Arg Thr Ser
785 790 795 800
tca gtc gag gtt tcg ctt aaa ttg aaa aac act ggg aaa aaa gct gga 2448
Ser Val Glu Val Ser Leu Lys Leu Lys Asn Thr Gly Lys Lys Ala Gly
805 810 815
gct gag gtt gta cag gta tac agc ggt aag ctc ccg acc aat gta gag 2496
Ala Glu Val Val Gln Val Tyr Ser Gly Lys Leu Pro Thr Asn Val Glu
820 825 830
acc cca tcc cgt caa ctg gca ggt tgg gcc aag gtg gaa tta aag ccg 2544
Thr Pro Ser Arg Gln Leu Ala Gly Trp Ala Lys Val Glu Leu Lys Pro
835 840 845
ggc caa gag aag aaa gtc cgc att gag ctt gac ccg aaa gct ctc tct 2592
Gly Gln Glu Lys Lys Val Arg Ile Glu Leu Asp Pro Lys Ala Leu Ser
850 855 860
tat tgg gat gaa aaa tcc aaa gca tgg gtt atg cca tct ggt gaa gtt 2640
Tyr Trp Asp Glu Lys Ser Lys Ala Trp Val Met Pro Ser Gly Glu Val
865 870 875 880
ccg att tat gta ggc agc tcc tct cag gat aca aga ctg aca gga agt 2688
Pro Ile Tyr Val Gly Ser Ser Ser Gln Asp Thr Arg Leu Thr Gly Ser
885 890 895
gtt acg att ccg gca acc tct aca gaa aag gca aaa aag taa 2730
Val Thr Ile Pro Ala Thr Ser Thr Glu Lys Ala Lys Lys
900 905
<210> 2
<211> 909
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Construct
<400> 2
Val Lys Val Lys Ser Thr Trp Leu Phe Arg Met Val Met Met Thr Val
1 5 10 15
Ile Ala Ala Val Val Ile Gly Pro Met His Ile Ala Gly Ala Ala Gly
20 25 30
Ser Arg Thr Asp Arg Pro Trp Met Asn Thr Ser Leu Ser Ala Glu Lys
35 40 45
Arg Thr Ala Leu Leu Leu Gln Glu Met Thr Leu Glu Glu Lys Ile Asp
50 55 60
Leu Val Thr Gly Lys Val Asn Asn Tyr Tyr Gly Phe Tyr Asn Asn Ser
65 70 75 80
Met Glu Arg Leu Gly Ile Pro Ala Leu Lys Met Ala Asp Gly Pro Ala
85 90 95
Gly Val Arg Ile Ala Asn Pro Asp Val Gln Asp Lys Gln Ser Thr Ala
100 105 110
Leu Pro Ala Pro Ile Ala Leu Ala Ala Thr Trp Asp Thr Gln Ala Ala
115 120 125
Lys Gln Tyr Gly Asp Leu Leu Gly Asp Glu Ala Phe Asn Thr Thr His
130 135 140
Asn Val Val Leu Gly Pro Gly Met Asp Ile Ala Arg Ile Pro Trp Gly
145 150 155 160
Ser Arg Asn Phe Glu Ser Met Gly Glu Asp Pro Leu Leu Gln Ser Gln
165 170 175
Met Ala Thr Ala Tyr Val Lys Gly Val Gln Ser His Pro Val Leu Ala
180 185 190
Thr Ala Lys His Tyr Leu Met Asn Asn Gln Glu Thr Glu Arg Phe Thr
195 200 205
Thr Asn Val Lys Val Ser Asp Arg Ala Leu His Glu Ile Tyr Ile Arg
210 215 220
Pro Phe Glu Glu Ala Ile Ala Lys Ala Asp Leu Gly Gly Ala Met Cys
225 230 235 240
Ser Phe Asn Lys Val Asn Gly Glu Ser Ala Cys Glu Asn Lys Thr Ile
245 250 255
Leu Thr Asp Leu Leu Lys Lys Glu Met Gln Phe Lys Gly Phe Val Met
260 265 270
Ser Asp Tyr Gly Ala Asn Leu Ser Thr Val Glu Ser Ala Asn Ser Gly
275 280 285
Leu Asp Leu Glu Thr Pro Gly Thr Pro Tyr Gly Lys Trp Gly Asp Gln
290 295 300
Leu Leu Asp Ala Val Lys Thr Gly Lys Val Ser Glu Gln Thr Ile Asp
305 310 315 320
Asp Lys Ala Lys Arg Ile Leu Val Gln Met Phe Ser Lys Gly Leu Phe
325 330 335
Asp His Pro Ala Gln Asn Asn Gln Ile Asp Ala Arg Asp His Gly Lys
340 345 350
Thr Ala Arg Gln Leu Ala Glu Glu Ser Met Val Leu Leu Gln Asn Lys
355 360 365
Asn Asn Val Leu Pro Leu Ser Gln Asp Lys Leu Lys Ser Ile Ala Val
370 375 380
Ile Gly Pro Asp Ala Asp Asn Gly Thr Val Ala Gly Gly Gly Ser Ser
385 390 395 400
Leu Val Asn Pro Thr Tyr Thr Val Ser Pro Leu Glu Gly Ile Arg Asn
405 410 415
Arg Val Gly Lys Gly Val Thr Val Gln Tyr Ala Pro Gly Thr Asp Pro
420 425 430
Ile Ser Ala Gly Asp Ile Met Pro Gly Pro Ser Ala Val Pro Ser Ser
435 440 445
Leu Leu Thr Thr Ser Asp Gln Lys Glu Asn Ile Ser Val Gly Tyr Ala
450 455 460
Thr Tyr Gly Asp Ala Glu Gln Gly Leu Arg Gly Glu Tyr Trp Lys Asn
465 470 475 480
Asn Lys Met Glu Gly Asn Pro Ile Leu Val Arg Asn Asp Asp Gln Val
485 490 495
Asn Met Asn Leu Gly Phe Tyr Asn Tyr Gln Gly Phe Asn Ala Lys Ser
500 505 510
Pro Lys Val Pro Asn Thr Pro Thr Thr Leu Asn Gly Leu Ile Ser Ala
515 520 525
Arg Trp Thr Gly Ala Ile Ala Ala Pro Lys Asp Gly Asp Tyr Ala Leu
530 535 540
Ser Leu Thr Ser Leu Gly Ser Ser Lys Leu Tyr Leu Asp Asp Lys Leu
545 550 555 560
Phe Val Asp Asn Gln Gly Thr Lys Leu Glu Thr Thr Lys Lys Asn Ile
565 570 575
Ser Phe Lys Ala Gly Glu Lys His Lys Ile Arg Ile Glu Tyr Arg Ala
580 585 590
Asp Tyr Pro Thr Asn Gly Arg Asp Ser Gly Gly Met Val Arg Leu Gly
595 600 605
Trp Glu Pro Pro Ala Asp Thr Thr Asp Lys Leu Ile Asp Asn Ala Val
610 615 620
Lys Leu Ala Lys Lys Ser Asp Val Ala Ile Val Val Thr Arg Thr Tyr
625 630 635 640
Glu Ser Glu Gly Tyr Val Glu Arg Ser Asp Met Glu Leu Pro Asn Asn
645 650 655
Gln Asp Arg Leu Ile Arg Ala Val Ala Ala Ala Asn Pro Lys Thr Ile
660 665 670
Val Val Gln Met Ser Gly Arg Ala Val Gln Met Asp Thr Trp Gln Asp
675 680 685
Lys Val Pro Ala Ile Val Gln Ala Trp Phe Ala Gly Gln Glu Gln Gly
690 695 700
Asn Ala Ile Ala Arg Val Leu Phe Gly Asp Val Asn Pro Ser Gly Lys
705 710 715 720
Leu Pro Val Thr Phe Pro Val Asn Glu Gln Ser Thr Pro Val Ser Ser
725 730 735
Pro Glu Lys Phe Pro Gly Val Asn Gly Val Gly Asp Tyr Ser Asp Gly
740 745 750
Ile Phe Val Gly Tyr Arg Gly Tyr Glu Lys Ser Gly Ile Gln Pro Ala
755 760 765
Phe Ser Phe Gly His Gly Leu Ser Tyr Thr Thr Phe Gly Tyr Ser Asp
770 775 780
Leu Lys Val Lys Gln His Ala Ser Gly Lys Lys Ser Asp Arg Thr Ser
785 790 795 800
Ser Val Glu Val Ser Leu Lys Leu Lys Asn Thr Gly Lys Lys Ala Gly
805 810 815
Ala Glu Val Val Gln Val Tyr Ser Gly Lys Leu Pro Thr Asn Val Glu
820 825 830
Thr Pro Ser Arg Gln Leu Ala Gly Trp Ala Lys Val Glu Leu Lys Pro
835 840 845
Gly Gln Glu Lys Lys Val Arg Ile Glu Leu Asp Pro Lys Ala Leu Ser
850 855 860
Tyr Trp Asp Glu Lys Ser Lys Ala Trp Val Met Pro Ser Gly Glu Val
865 870 875 880
Pro Ile Tyr Val Gly Ser Ser Ser Gln Asp Thr Arg Leu Thr Gly Ser
885 890 895
Val Thr Ile Pro Ala Thr Ser Thr Glu Lys Ala Lys Lys
900 905
Claims (5)
1.一种β-葡萄糖苷酶基因，其特征在于，核苷酸序列如SEQ ID NO：1所示。
2.一种重组载体，其特征在于，所述重组载体含有权利要求1所述的β-葡萄糖苷酶基因。
3.一种宿主细胞，其特征在于，所述宿主细胞含有权利要求2所述的重组载体。
4.根据权利要求3所述的宿主细胞，其特征在于，所述宿主细胞为原核细胞。
5.根据权利要求4所述的宿主细胞，其特征在于，所述原核细胞为大肠杆菌E.coli BL21(DE3)。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610424102.5A CN105969751B (zh) | 2016-06-14 | 2016-06-14 | 一种β-葡萄糖苷酶基因及其应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610424102.5A CN105969751B (zh) | 2016-06-14 | 2016-06-14 | 一种β-葡萄糖苷酶基因及其应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105969751A true CN105969751A (zh) | 2016-09-28 |
| CN105969751B CN105969751B (zh) | 2019-12-31 |
Family
ID=57021857
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610424102.5A Active CN105969751B (zh) | 2016-06-14 | 2016-06-14 | 一种β-葡萄糖苷酶基因及其应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105969751B (zh) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111004792A (zh) * | 2020-01-06 | 2020-04-14 | 陈炜山 | 一种高耐受型β-葡萄糖苷酶的制备方法及其应用 |
| CN114250316A (zh) * | 2021-12-22 | 2022-03-29 | 绍兴市水产技术推广中心 | 一种从养殖塘水土样本中检测虾肝肠胞虫的方法 |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102321647A (zh) * | 2011-09-08 | 2012-01-18 | 杭州师范大学 | 一种β-葡萄糖苷酶、编码基因、载体、工程菌及其应用 |
| CN103114100A (zh) * | 2013-02-27 | 2013-05-22 | 广西科学院 | β-葡萄糖苷酶基因S-bgl3及其应用 |
| CN103881958A (zh) * | 2014-03-31 | 2014-06-25 | 上海交通大学 | 基于β-葡萄糖苷酶的工程菌及其实现方法 |
| CN109072209A (zh) * | 2016-03-24 | 2018-12-21 | 诺维信公司 | 纤维二糖水解酶变体和编码其的多核苷酸 |
| CN109112118A (zh) * | 2011-11-21 | 2019-01-01 | 诺维信股份有限公司 | Gh61多肽变体以及编码所述变体的多核苷酸 |
-
2016
- 2016-06-14 CN CN201610424102.5A patent/CN105969751B/zh active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102321647A (zh) * | 2011-09-08 | 2012-01-18 | 杭州师范大学 | 一种β-葡萄糖苷酶、编码基因、载体、工程菌及其应用 |
| CN109112118A (zh) * | 2011-11-21 | 2019-01-01 | 诺维信股份有限公司 | Gh61多肽变体以及编码所述变体的多核苷酸 |
| CN103114100A (zh) * | 2013-02-27 | 2013-05-22 | 广西科学院 | β-葡萄糖苷酶基因S-bgl3及其应用 |
| CN103881958A (zh) * | 2014-03-31 | 2014-06-25 | 上海交通大学 | 基于β-葡萄糖苷酶的工程菌及其实现方法 |
| CN109072209A (zh) * | 2016-03-24 | 2018-12-21 | 诺维信公司 | 纤维二糖水解酶变体和编码其的多核苷酸 |
Non-Patent Citations (5)
| Title |
|---|
| GANG LI等: "Molecular cloning and characterization of a novel b-glucosidase with high hydrolyzing ability for soybean isoflavone glycosides and glucose-tolerance from soil metagenomic library", 《BIORESOURCE TECHNOLOGY》 * |
| NCBI DATABASE: "ACCESSION WP_016819903", 《GENBANK》 * |
| WEILIANG DONG等: "Characterization of a β-glucosidase from Paenibacillus species and its application for succinic acid production from sugarcane bagasse hydrolysate.", 《BIORESOUR TECHNOL.》 * |
| 徐蓉等: "利用纤维素水解液中的纤维二糖发酵制备丁二酸", 《化工进展》 * |
| 李泰明等: "黑曲霉B-葡萄糖苷酶基因在大肠杆菌中的克隆与表达", 《安徽医药》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111004792A (zh) * | 2020-01-06 | 2020-04-14 | 陈炜山 | 一种高耐受型β-葡萄糖苷酶的制备方法及其应用 |
| CN114250316A (zh) * | 2021-12-22 | 2022-03-29 | 绍兴市水产技术推广中心 | 一种从养殖塘水土样本中检测虾肝肠胞虫的方法 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105969751B (zh) | 2019-12-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105671070B (zh) | 一种用于枯草芽孢杆菌基因组编辑的CRISPRCas9***及其构建方法 | |
| CN104974944B (zh) | 一种产dha的裂殖壶菌基因工程菌及其构建方法和应用 | |
| Golotin et al. | Optimization of cold-adapted alpha-galactosidase expression in Escherichia coli | |
| CN104152505A (zh) | 一种利用重组菌株转化制备4-羟基-l-异亮氨酸的方法 | |
| CN107603937A (zh) | 一种表达赖氨酸氨肽酶的重组大肠杆菌及其构建方法 | |
| CN109880813A (zh) | 一种具有低聚半乳糖合成能力的β-葡萄糖苷酶及其表达菌株和应用 | |
| CN108841770A (zh) | 一种能够表达磷脂酶d的枯草芽孢杆菌工程菌 | |
| CN104046586B (zh) | 一株基因工程菌及其在生产(2r,3r)-2,3-丁二醇中的应用 | |
| CN107881140A (zh) | 一株高产甘露醇的肠膜明串珠菌突变菌株及其应用方法 | |
| CN105969751A (zh) | 一种β-葡萄糖苷酶基因及其应用 | |
| CN104845926B (zh) | 一种有利于重组蛋白胞外分泌的基因敲除大肠杆菌及其应用 | |
| CN106434715B (zh) | 麦芽寡糖基海藻糖合成酶及其表达基因与应用 | |
| CN106434587B (zh) | 一种葡聚糖蔗糖酶及其应用 | |
| CN109022396A (zh) | 一种酶活提高的α-淀粉酶突变体及其应用 | |
| CN108660121A (zh) | 一种热稳定性提高的淀粉分支酶突变体 | |
| CN108220216B (zh) | 一种过表达glnR基因的耐铵固氮微生物及其构建方法与应用 | |
| CN105969713A (zh) | 一种高产麦芽寡糖基海藻糖水解酶的基因工程菌及其应用 | |
| CN101250539B (zh) | 一种制备重组耐热β-葡萄糖醛酸酶的方法 | |
| CN106520729B (zh) | 麦芽寡糖基海藻糖水解酶及其表达基因与应用 | |
| Vu et al. | Improvement of cellulase activity using error-prone rolling circle amplification and site-directed mutagenesis | |
| CN111549050B (zh) | 适用于芽胞杆菌的透明颤菌血红蛋白表达框及应用 | |
| CN103966185A (zh) | 高效合成s-腺苷同型半胱氨酸的双酶体系及其应用方法 | |
| CN106434586A (zh) | 海藻糖合成酶突变体及其基因 | |
| CN104651332B (zh) | 嗜热细菌海藻糖合酶c端片段提高海藻糖合酶酶活的方法 | |
| US10465177B2 (en) | Maltooligosyl trehalose trehalohydrolase (MTHase) mutant and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20240628 Address after: 518118 Innovation Square A1301, 2007 Pingshan Avenue, Liulian Community, Pingshan Street, Pingshan District, Shenzhen City, Guangdong Province Patentee after: Xiangsheng New Materials (Shenzhen) Co.,Ltd. Country or region after: China Address before: 211816 601, Building A, Nanjing Tech University, No. 30 Puzhu South Road, Pukou District, Nanjing City, Jiangsu Province Patentee before: Nanjing Tech University Country or region before: China |