JP2020506704A5 - - Google Patents

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Publication number

JP2020506704A5

JP2020506704A5 JP2019541715A JP2019541715A JP2020506704A5 JP 2020506704 A5 JP2020506704 A5 JP 2020506704A5 JP 2019541715 A JP2019541715 A JP 2019541715A JP 2019541715 A JP2019541715 A JP 2019541715A JP 2020506704 A5 JP2020506704 A5 JP 2020506704A5

Authority

JP

Japan

Prior art keywords

glycosyltransferase

engineered

item

positions

seq

Prior art date

2018-02-01

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

• Espacenet
• Global Dossier
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• 108700023372 Glycosyltransferases Proteins 0.000 claims description 191
• 102000051366 Glycosyltransferases Human genes 0.000 claims description 187
• 230000035772 mutation Effects 0.000 claims description 134
• 229920001184 polypeptide Polymers 0.000 claims description 71
• 102000004196 processed proteins & peptides Human genes 0.000 claims description 71
• 108090000765 processed proteins & peptides Proteins 0.000 claims description 71
• 238000000034 method Methods 0.000 claims description 67
• 108010043934 Sucrose synthase Proteins 0.000 claims description 61
• 235000019202 steviosides Nutrition 0.000 claims description 55
• UEDUENGHJMELGK-HYDKPPNVSA-N Stevioside Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O UEDUENGHJMELGK-HYDKPPNVSA-N 0.000 claims description 53
• 229940013618 stevioside Drugs 0.000 claims description 53
• OHHNJQXIOPOJSC-UHFFFAOYSA-N stevioside Natural products CC1(CCCC2(C)C3(C)CCC4(CC3(CCC12C)CC4=C)OC5OC(CO)C(O)C(O)C5OC6OC(CO)C(O)C(O)C6O)C(=O)OC7OC(CO)C(O)C(O)C7O OHHNJQXIOPOJSC-UHFFFAOYSA-N 0.000 claims description 53
• 239000000758 substrate Substances 0.000 claims description 52
• 229930006000 Sucrose Natural products 0.000 claims description 26
• CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 26
• 108700014210 glycosyltransferase activity proteins Proteins 0.000 claims description 26
• 239000005720 sucrose Substances 0.000 claims description 26
• 239000008103 glucose Substances 0.000 claims description 21
• 238000004519 manufacturing process Methods 0.000 claims description 20
• -1 NDP Substances 0.000 claims description 16
• 239000000203 mixture Substances 0.000 claims description 14
• WFPZSXYXPSUOPY-ROYWQJLOSA-N ADP alpha-D-glucoside Chemical compound C([C@H]1O[C@H]([C@@H]([C@@H]1O)O)N1C=2N=CN=C(C=2N=C1)N)OP(O)(=O)OP(O)(=O)O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O WFPZSXYXPSUOPY-ROYWQJLOSA-N 0.000 claims description 13
• WFPZSXYXPSUOPY-UHFFFAOYSA-N ADP-mannose Natural products C1=NC=2C(N)=NC=NC=2N1C(C(C1O)O)OC1COP(O)(=O)OP(O)(=O)OC1OC(CO)C(O)C(O)C1O WFPZSXYXPSUOPY-UHFFFAOYSA-N 0.000 claims description 13
• 230000001419 dependent effect Effects 0.000 claims description 12
• 102000040430 polynucleotide Human genes 0.000 claims description 9
• 108091033319 polynucleotide Proteins 0.000 claims description 9
• 239000002157 polynucleotide Substances 0.000 claims description 9
• RPYRMTHVSUWHSV-CUZJHZIBSA-N rebaudioside D Chemical compound O([C@H]1[C@H](O)[C@@H](CO)O[C@H]([C@@H]1O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O RPYRMTHVSUWHSV-CUZJHZIBSA-N 0.000 claims description 8
• 239000000047 product Substances 0.000 claims description 7
• 230000013595 glycosylation Effects 0.000 claims description 5
• 238000006206 glycosylation reaction Methods 0.000 claims description 5
• MVMSCBBUIHUTGJ-UHFFFAOYSA-N 10108-97-1 Natural products C1=2NC(N)=NC(=O)C=2N=CN1C(C(C1O)O)OC1COP(O)(=O)OP(O)(=O)OC1OC(CO)C(O)C(O)C1O MVMSCBBUIHUTGJ-UHFFFAOYSA-N 0.000 claims description 4
• CGPHZDRCVSLMCF-JZMIEXBBSA-N CDP-alpha-D-glucose Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)O[C@@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)O1 CGPHZDRCVSLMCF-JZMIEXBBSA-N 0.000 claims description 4
• 229930091371 Fructose Natural products 0.000 claims description 4
• 239000005715 Fructose Substances 0.000 claims description 4
• RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 claims description 4
• MVMSCBBUIHUTGJ-LRJDVEEWSA-N GDP-alpha-D-glucose Chemical compound C([C@H]1O[C@H]([C@@H]([C@@H]1O)O)N1C=2N=C(NC(=O)C=2N=C1)N)OP(O)(=O)OP(O)(=O)O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O MVMSCBBUIHUTGJ-LRJDVEEWSA-N 0.000 claims description 4
• 239000007795 chemical reaction product Substances 0.000 claims description 4
• YSYKRGRSMLTJNL-URARBOGNSA-N dTDP-alpha-D-glucose Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)O[C@@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)[C@@H](O)C1 YSYKRGRSMLTJNL-URARBOGNSA-N 0.000 claims description 4
• 241000894006 Bacteria Species 0.000 claims description 3
• 241000206602 Eukaryota Species 0.000 claims description 3
• 238000012258 culturing Methods 0.000 claims description 3
• 241001247437 Cerbera odollam Species 0.000 claims description 2
• 244000228451 Stevia rebaudiana Species 0.000 claims description 2
• 235000006092 Stevia rebaudiana Nutrition 0.000 claims description 2
• 239000004383 Steviol glycoside Substances 0.000 claims description 2
• HSCJRCZFDFQWRP-JZMIEXBBSA-N UDP-alpha-D-glucose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OP(O)(=O)OP(O)(=O)OC[C@@H]1[C@@H](O)[C@@H](O)[C@H](N2C(NC(=O)C=C2)=O)O1 HSCJRCZFDFQWRP-JZMIEXBBSA-N 0.000 claims description 2
• 238000006243 chemical reaction Methods 0.000 claims description 2
• 238000004140 cleaning Methods 0.000 claims description 2
• 238000004440 column chromatography Methods 0.000 claims description 2
• 238000005580 one pot reaction Methods 0.000 claims description 2
• RAYIFMWTQKNDNK-UHFFFAOYSA-N phosphono dihydrogen phosphate;1h-pyrimidine-2,4-dione Chemical compound O=C1C=CNC(=O)N1.OP(O)(=O)OP(O)(O)=O RAYIFMWTQKNDNK-UHFFFAOYSA-N 0.000 claims description 2
• GSGVXNMGMKBGQU-PHESRWQRSA-N rebaudioside M Chemical compound C[C@@]12CCC[C@](C)([C@H]1CC[C@@]13CC(=C)[C@@](C1)(CC[C@@H]23)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O[C@@H]2O[C@H](CO)[C@@H](O)[C@H](O)[C@H]2O)[C@H]1O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O)C(=O)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O[C@@H]2O[C@H](CO)[C@@H](O)[C@H](O)[C@H]2O)[C@H]1O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O GSGVXNMGMKBGQU-PHESRWQRSA-N 0.000 claims description 2
• 229930182488 steviol glycoside Natural products 0.000 claims description 2
• 235000019411 steviol glycoside Nutrition 0.000 claims description 2
• 150000008144 steviol glycosides Chemical class 0.000 claims description 2
• 102000045442 glycosyltransferase activity proteins Human genes 0.000 claims 4
• 230000015572 biosynthetic process Effects 0.000 claims 1
• 229930188195 rebaudioside Natural products 0.000 claims 1
• 238000003786 synthesis reaction Methods 0.000 claims 1
• 239000001512 FEMA 4601 Substances 0.000 description 3
• HELXLJCILKEWJH-SEAGSNCFSA-N Rebaudioside A Natural products O=C(O[C@H]1[C@@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1)[C@@]1(C)[C@@H]2[C@](C)([C@H]3[C@@]4(CC(=C)[C@@](O[C@H]5[C@H](O[C@H]6[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O6)[C@@H](O[C@H]6[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O6)[C@H](O)[C@@H](CO)O5)(C4)CC3)CC2)CCC1 HELXLJCILKEWJH-SEAGSNCFSA-N 0.000 description 3
• HELXLJCILKEWJH-UHFFFAOYSA-N entered according to Sigma 01432 Natural products C1CC2C3(C)CCCC(C)(C(=O)OC4C(C(O)C(O)C(CO)O4)O)C3CCC2(C2)CC(=C)C21OC(C1OC2C(C(O)C(O)C(CO)O2)O)OC(CO)C(O)C1OC1OC(CO)C(O)C(O)C1O HELXLJCILKEWJH-UHFFFAOYSA-N 0.000 description 3
• 125000003147 glycosyl group Chemical group 0.000 description 3
• HELXLJCILKEWJH-NCGAPWICSA-N rebaudioside A Chemical compound O([C@H]1[C@H](O)[C@@H](CO)O[C@H]([C@@H]1O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O HELXLJCILKEWJH-NCGAPWICSA-N 0.000 description 3
• 235000019203 rebaudioside A Nutrition 0.000 description 3
• 102000004357 Transferases Human genes 0.000 description 2
• 108090000992 Transferases Proteins 0.000 description 2
• FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
• 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 1
• HSCJRCZFDFQWRP-UHFFFAOYSA-N Uridindiphosphoglukose Natural products OC1C(O)C(O)C(CO)OC1OP(O)(=O)OP(O)(=O)OCC1C(O)C(O)C(N2C(NC(=O)C=C2)=O)O1 HSCJRCZFDFQWRP-UHFFFAOYSA-N 0.000 description 1
• 230000001279 glycosylating effect Effects 0.000 description 1