公开(公告)号：US12065685B1

公开(公告)日：2024-08-20

申请号：US18368369

申请日：2023-09-14

申请人： AMYRIS, INC.

发明人： Sean Lund ,  Gale Wichmann

IPC分类号： C12P19/56 ,  C12N15/81

CPC分类号： C12P19/56 ,  C12N15/81 ,  C12Y106/02004 ,  C12Y114/14 ,  C12Y205/01029 ,  C12Y402/03019 ,  C12Y505/01012

摘要： Provided herein are genetically modified host cells, compositions, and methods for improved production of steviol glycosides. The host cells are genetically modified to contain a heterologous nucleic acid that expresses novel and optimized variants of UGT76G1. The host cell further contains one or more heterologous nucleotide sequence encoding further enzymes of a pathway capable of producing one or more steviol glycosides in the host cell. The host cells, compositions, and methods described herein provide an efficient route for the heterologous production of rebaudioside M.

公开(公告)号：US20160102331A1

公开(公告)日：2016-04-14

申请号：US14893284

申请日：2014-06-02

申请人： DSM IP ASSETS B.V.

发明人： Viktor Marius BOER ,  Nicolette Jasmijn BROERS ,  Adam G. LAWRENCE

IPC分类号： C12P19/56 ,  C12N9/90 ,  A23L2/60 ,  C12N9/02 ,  A23L1/236 ,  A23K1/16 ,  C07H15/256 ,  C12N9/88

CPC分类号： C12P19/56 ,  A23K20/105 ,  A23L2/60 ,  A23L27/36 ,  A23V2002/00 ,  C07H15/256 ,  C12N9/0042 ,  C12N9/0073 ,  C12N9/0083 ,  C12N9/1051 ,  C12N9/88 ,  C12N9/90 ,  C12N15/52 ,  C12P15/00 ,  C12Y114/13 ,  C12Y114/13078 ,  C12Y402/03019 ,  C12Y505/01013

摘要： The present invention relates to a method for the production of a diterpene or a glycosylated diterpene, which method comprises: a. fermenting a recombinant microorganism in a suitable fermentation medium, wherein the microorganism comprises one or more nucleotide sequence(s) encoding: a polypeptide having ent-copalyl pyrophosphate synthase activity; a polypeptide having ent-Kaurene synthase activity; a polypeptide having ent-Kaurene oxidase activity; and a polypeptide having kaurenoic acid 13-hydroxylase activity and whereby expression of the nucleotide sequence(s) confer(s) on the microorganism the ability to produce at least steviol, whereby a diterpene or glycosylated diterpene is produced extracellularly in the fermentation medium; and b. recovering the diterpene or glycosylated diterpene from the fermentation medium.

摘要翻译： 本发明涉及二萜或糖基二萜的制备方法，该方法包括：a。 在合适的发酵培养基中发酵重组微生物，其中所述微生物包含一个或多个核苷酸序列，所述核苷酸序列编码：具有入 - copalyl焦磷酸合酶活性的多肽; 具有ent-Kaurene合酶活性的多肽; 具有ent-Kaurene氧化酶活性的多肽; 和具有山梨酸13-羟化酶活性的多肽，由此所述核苷酸序列的表达赋予所述微生物至少产生甜菊醇的能力，从而在发酵培养基中细胞外产生二萜或糖基化二萜; 和b。 从发酵培养基中回收二萜或糖基化二萜。

公开(公告)号：US20190078128A9

公开(公告)日：2019-03-14

申请号：US15382354

申请日：2016-12-16

申请人： EVOLVA SA

发明人： Ganesh M. KISHORE ,  Michael MOTION ,  Paula M. HICKS ,  Jorgen HANSEN ,  Jens HOUGHTON-LARSEN ,  Esben Halkjaer HANSEN ,  Michael Dalgaard MIKKELSEN ,  Sabina TAVARES ,  Charlotte BLOM

IPC分类号： C12P19/56 ,  C07H15/256 ,  C12N9/04 ,  C12N9/10 ,  C12N9/90 ,  C12N9/88 ,  C12N9/02 ,  A23L27/30 ,  A23L2/60 ,  A23L33/10

CPC分类号： C12P19/56 ,  A23L2/60 ,  A23L27/36 ,  C07H15/256 ,  C12N9/0006 ,  C12N9/0042 ,  C12N9/0073 ,  C12N9/1085 ,  C12N9/88 ,  C12N9/90 ,  C12P15/00 ,  C12Y101/01088 ,  C12Y106/02004 ,  C12Y114/13078 ,  C12Y114/13079 ,  C12Y205/01029 ,  C12Y402/03019 ,  C12Y505/01012

摘要： Recombinant microorganisms, plants, and plant cells are disclosed that have been engineered to express novel recombinant genes encoding steviol biosynthetic enzymes and UDP-glycosyltransferases (UGTs). Such microorganisms, plants, or plant cells can produce steviol or steviol glycosides, e.g., rubusoside or Rebaudioside A, which can be used as natural sweeteners in food products and dietary supplements.

公开(公告)号：US20170306377A1

公开(公告)日：2017-10-26

申请号：US15516854

申请日：2015-10-08

申请人： DSM IP ASSETS B.V.

发明人： Marco Alexander VAN DEN BERG ,  Marcella Katharina MADERN ,  Remko Tsjibbe WINTER

IPC分类号： C12P19/56 ,  C12N15/52 ,  A23L27/30

CPC分类号： C12P19/56 ,  A23L27/33 ,  A23L27/36 ,  A23V2002/00 ,  C12N15/52 ,  C12P19/18 ,  C12Y106/02004 ,  C12Y114/13078 ,  C12Y114/13079 ,  C12Y204/00 ,  C12Y402/03019 ,  C12Y505/01013

摘要： The present invention relates to a method for increasing the degree of glycosylation of a composition comprising steviol glycosides, which method comprises:a. contacting said composition comprising steviol glycosides with a recombinant microorganism, a cell free extract derived from such a microorganism or an enzyme preparation derived from either thereof; and b. thereby to increase the degree of glycosylation of the composition comprising steviol glycosides, wherein the recombinant microorganism comprises one or more nucleotide sequence(s) encoding: a polypeptide having ent-copalyl pyrophosphate synthase activity;a polypeptide having ent-Kaurene synthase activity; a polypeptide having ent-Kaurene oxidase activity; a polypeptide having kaurenoic acid 13-hydroxylase activity; and one or more polypeptides having UDP-glucosyltransferase activity whereby expression of the nucleotide sequence(s) confer(s) on the microorganism the ability to produce at least one steviol glycoside. The present invention also relates to a composition comprising steviol glycosides obtainable by such a method.

公开(公告)号：US20230146484A1

公开(公告)日：2023-05-11

申请号：US17981671

申请日：2022-11-07

申请人： MANUS BIO INC.

发明人： Ryan PHILIPPE ,  Ajikumar PARAYIL KUMARAN ,  Christine Nicole S. SANTOS

IPC分类号： C12P19/56

CPC分类号： C12P19/56 ,  C12N1/205 ,  C12Y114/14 ,  C12Y114/13079 ,  C12Y204/01 ,  C12Y402/03019

摘要： The present invention provides engineered cells and methods for making high purity steviol glycosides, including RebM. In some aspects, the present invention provides host cells, such as bacterial cells (including but not limited to E. coli), that are engineered to overexpress and/or delete or inactivate one or more steviol glycoside transport proteins. The bacterial cells selectively export RebM, or other specific combination of steviol glycosides, out of the cell to increase productivity and reduce production costs associated with downstream purification. Non-target steviol glycosides are not transported to the extracellular medium in significant amounts.

公开(公告)号：US10017804B2

公开(公告)日：2018-07-10

申请号：US14764898

申请日：2014-02-11

申请人： Evolva SA

发明人： Ernesto Simon ,  Iben Nordmark Anderson ,  Michael Dalgaard Mikkelsen ,  Jorgen Hansen ,  Veronique Douchin

IPC分类号： C12P19/56 ,  C12N9/02 ,  C07K14/395 ,  C12N9/10 ,  C12P15/00 ,  C12N9/88 ,  C12N9/90

CPC分类号： C12P19/56 ,  C07K14/395 ,  C12N9/0042 ,  C12N9/0073 ,  C12N9/1048 ,  C12N9/1051 ,  C12N9/1062 ,  C12N9/1085 ,  C12N9/88 ,  C12N9/90 ,  C12P15/00 ,  C12Y106/02004 ,  C12Y114/13078 ,  C12Y204/00 ,  C12Y204/01013 ,  C12Y204/01035 ,  C12Y205/01029 ,  C12Y402/03 ,  C12Y402/03019 ,  C12Y505/01013

摘要： Recombinant microorganisms are disclosed that produce steviol glycosides and have altered expression of one or more endogenous transporter or transcription factor genes, or that overexpress one or more heterologous transporters, leading to increased excretion of steviol glycosides of interest.

公开(公告)号：US20170218419A1

公开(公告)日：2017-08-03

申请号：US15382354

申请日：2016-12-16

申请人： EVOLVA SA

发明人： Ganesh M. KISHORE ,  Michael MOTION ,  Paula M. HICKS ,  Jorgen HANSEN ,  Jens HOUGHTON-LARSEN ,  Esben Halkjaer HANSEN ,  Michael Dalgaard MIKKELSEN ,  Sabina TAVARES ,  Charlotte BLOM

IPC分类号： C12P19/56 ,  C12N9/04 ,  C12N9/10 ,  A23L33/10 ,  C12N9/88 ,  C12N9/02 ,  A23L27/30 ,  A23L2/60 ,  C07H15/256 ,  C12N9/90

CPC分类号： C12P19/56 ,  A23L2/60 ,  A23L27/36 ,  C07H15/256 ,  C12N9/0006 ,  C12N9/0042 ,  C12N9/0073 ,  C12N9/1085 ,  C12N9/88 ,  C12N9/90 ,  C12P15/00 ,  C12Y101/01088 ,  C12Y106/02004 ,  C12Y114/13078 ,  C12Y114/13079 ,  C12Y205/01029 ,  C12Y402/03019 ,  C12Y505/01012

摘要： Recombinant microorganisms, plants, and plant cells are disclosed that have been engineered to express novel recombinant genes encoding steviol biosynthetic enzymes and UDP-glycosyltransferases (UGTs). Such microorganisms, plants, or plant cells can produce steviol or steviol glycosides, e.g., rubusoside or Rebaudioside A, which can be used as natural sweeteners in food products and dietary supplements.

公开(公告)号：US20230212630A1

公开(公告)日：2023-07-06

申请号：US17899069

申请日：2022-08-30

申请人： EVOLVA SA

发明人： Helene Lunde Robertson ,  Iben Nordmark Andersen ,  Adam Matthew Takos ,  Swee Chuang Lim Hallwyl ,  Francesca Ambri ,  Manuel Quiros Asensio ,  Michael Dalgaard Mikkelsen ,  Jens Houghton-Larsen ,  Veronique Douchin ,  Jane Dannow Dyekjaer ,  Simon Carlsen ,  Nina Nicoline Rasmussen ,  Esben Halkjaer Hansen

IPC分类号： C12P19/56 ,  A23L27/30 ,  C12N9/02 ,  C12N15/52 ,  C07H15/256 ,  C12N9/10 ,  C12N9/88 ,  C12N9/90

CPC分类号： C12P19/56 ,  A23L27/36 ,  C12Y114/13078 ,  C12N9/0042 ,  C12N9/0073 ,  C12N15/52 ,  C07H15/256 ,  C12N9/1085 ,  C12N9/88 ,  C12N9/90 ,  C12Y106/02004 ,  C12Y114/13079 ,  C12Y205/01029 ,  C12Y402/03019 ,  C12Y505/01013 ,  C07K2319/00 ,  A23V2002/00

摘要： The invention relates to recombinant microorganisms and methods for producing steviol glycosides and steviol glycoside precursors.

公开(公告)号：US20170240942A1

公开(公告)日：2017-08-24

申请号：US15506196

申请日：2015-09-09

申请人： EVOLVA SA

发明人： Helene Lunde Robertson ,  Iben Nordmark Andersen ,  Adam Matthew Takos ,  Swee Chuang Lim Hallwyl ,  Francesca Ambri ,  Manuel Quiros Asensio ,  Michael Dalgaard Mikkelsen ,  Jens Houghton-Larsen ,  Veronique Douchin ,  Jane Dannow Dyekjaer ,  Simon Carlsen ,  Nina Nicole Rasmussen ,  Esben Halkjaer Hansen

IPC分类号： C12P19/56 ,  C12N9/02 ,  A23L27/30 ,  C12N9/90 ,  C12N9/88 ,  C12N15/52 ,  C07H15/256 ,  C12N9/10

CPC分类号： C12P19/56 ,  A23L27/36 ,  A23V2002/00 ,  C07H15/256 ,  C07K2319/00 ,  C12N9/0042 ,  C12N9/0073 ,  C12N9/1085 ,  C12N9/88 ,  C12N9/90 ,  C12N15/52 ,  C12Y106/02004 ,  C12Y114/13078 ,  C12Y114/13079 ,  C12Y205/01029 ,  C12Y402/03019 ,  C12Y505/01013

摘要： The invention relates to recombinant microorganisms and methods for producing steviol glycosides and steviol glycoside precursors.

公开(公告)号：US20160160257A1

公开(公告)日：2016-06-09

申请号：US14906497

申请日：2014-07-23

申请人： DSM IP ASSETS B.V.

发明人： Nicolette Jasmijn BROERS ,  Viktor Marius BOER ,  Adam G. LAWRENCE

IPC分类号： C12P19/56 ,  C07H15/256 ,  C12N9/02 ,  C12N9/90 ,  C12N9/88 ,  C07C62/32 ,  C12P15/00

CPC分类号： C12P19/56 ,  A23L2/60 ,  A23L27/36 ,  C07C62/32 ,  C07C2603/86 ,  C07H15/256 ,  C12N9/0042 ,  C12N9/0073 ,  C12N9/0083 ,  C12N9/1051 ,  C12N9/88 ,  C12N9/90 ,  C12N15/52 ,  C12P15/00 ,  C12Y114/13078 ,  C12Y114/13079 ,  C12Y402/03019 ,  C12Y505/01013

摘要： The present invention relates to a method for the production of a diterpene or a glycosylated diterpene, which method comprises: a. fermenting a recombinant microorganism of the genus Yarrowia in a suitable fermentation medium at a temperature of about 29° C. or higher, wherein the microorganism comprises one or more nucleotide sequence(s) encoding: a polypeptide having ent-copalyl pyrophosphate synthase activity; a polypeptide having ent-Kaurene synthase activity; a polypeptide having ent-Kaurene oxidase activity; and a polypeptide having kaurenoic acid 13-hydroxylase activity and whereby expression of the nucleotide sequence(s) confer(s) on the microorganism the ability to produce at least steviol; and b. recovering the diterpene or glycosylated diterpene.

摘要翻译： 本发明涉及二萜或糖基二萜的制备方法，该方法包括：a。 在约29℃或更高的温度下在合适的发酵培养基中发酵耶氏酵母属的重组微生物，其中所述微生物包含一个或多个核苷酸序列，所述核苷酸序列编码：具有共 - 酰基焦磷酸合酶活性的多肽; 具有ent-Kaurene合酶活性的多肽; 具有ent-Kaurene氧化酶活性的多肽; 和具有山梨酸13-羟化酶活性的多肽，由此所述核苷酸序列的表达赋予所述微生物至少产生甜菊醇的能力; 和b。 回收二萜或糖基化二萜。