公开(公告)号：US11781121B2

公开(公告)日：2023-10-10

申请号：US17541565

申请日：2021-12-03

Applicant: DSM IP ASSETS B.V.

Inventor： Viktor Marius Boer ,  Priscilla Zwartjens ,  Johannes Gustaaf Ernst Van Leeuwen

IPC: C12P15/00 ,  C12P19/56 ,  C12N9/10 ,  C12N9/02 ,  C12N9/88 ,  C12N15/52 ,  C12P7/42

CPC classification number: C12N9/1085 ,  C12N9/0073 ,  C12N9/88 ,  C12N15/52 ,  C12P7/42 ,  C12P15/00 ,  C12P19/56 ,  C12Y114/13078 ,  C12Y205/01001 ,  C12Y402/03019

Abstract: The present invention relates a variant polypeptide having geranylgeranyl pyrophosphate synthase activity, which variant polypeptide comprises an amino acid sequence which, when aligned with a geranylgeranyl pyrophosphate synthase comprising the sequence set out in SEQ ID NO: 1, comprises at least one substitution of an amino acid residue corresponding to any of amino acids at positions 92, 100 or 235 said positions being defined with reference to SEQ ID NO: 1 and wherein the variant has one or more modified properties as compared with a reference polypeptide having geranylgeranyl pyrophosphate synthase activity. A variant polypeptide of the invention may be used in a recombinant host for the production of steviol or a steviol glycoside.

公开(公告)号：US11293043B2

公开(公告)日：2022-04-05

申请号：US16845736

申请日：2020-04-10

Applicant: DSM IP ASSETS B.V.

Inventor： Viktor Marius Boer ,  Priscilla Zwartjens ,  Eric Van Den Berg

IPC: C12N1/20 ,  C12P19/56 ,  C07K14/39 ,  C07H15/24 ,  C12N9/10

Abstract: A recombinant host capable of producing a steviol glycoside which overexpresses a polypeptide which mediates steviol glycoside transport and which polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 35 or SEQ ID NO: 38 or an amino acid sequence having at least about 50% sequence identity to either thereto. A recombinant host capable of producing a steviol glycoside which has been modified, preferably in its genome, to result in a deficiency in the production of a polypeptide which mediates steviol glycoside transport and which polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 35 or SEQ ID NO: 38 or an amino acid sequence having at least about 50% sequence identity to either thereto.

公开(公告)号：US11913034B2

公开(公告)日：2024-02-27

申请号：US17381479

申请日：2021-07-21

Applicant: DSM IP ASSETS B.V.

Inventor： Johannes Gustaaf Ernst Van Leeuwen ,  Priscilla Zwartjens ,  Viktor Marius Boer

IPC: C12N9/02 ,  A23L27/30 ,  C12P19/12

CPC classification number: C12N9/0073 ,  A23L27/36 ,  C12P19/12 ,  C12Y114/13079 ,  A23V2002/00

Abstract: The present invention relates a polypeptide having kaurenoic acid 13-hydroxylase activity, which polypeptide comprises an amino acid sequence which, when aligned with a kaurenoic acid 13-hydroxylase comprising the sequence set out in SEQ ID NO: 1 or SEQ ID NO: 3, comprises at least one substitution of an amino acid corresponding to any of amino acids at positions 136, 248, 336 or 403, said positions being defined with reference to SEQ ID NO: 1 or SEQ ID NO: 3 and wherein the polypeptide has one or more modified properties as compared with a reference polypeptide having kaurenoic acid 13-hydroxylase activity. A polypeptide of the invention may be used in a recombinant host for the production of steviol or a steviol glycoside.

公开(公告)号：US10793850B2

公开(公告)日：2020-10-06

申请号：US14384192

申请日：2013-03-12

Applicant: DSM IP ASSETS B.V.

Inventor： Nathalie Wiessenhaan ,  Catharina Petronella Antonia Maria Kolen ,  Bernard Meijrink ,  Viktor Marius Boer ,  Johannes Andries Roubos ,  Yvonne Johannes Odilia Arendsen

IPC: C12N15/90 ,  C12N15/10

Abstract: The present invention relates to a method for carrying out recombination at a target locus.

公开(公告)号：US10760085B2

公开(公告)日：2020-09-01

申请号：US15765549

申请日：2016-10-05

Applicant: DSM IP ASSETS B.V.

Inventor： Johannes Gustaaf Ernst Van Leeuwen ,  Priscilla Zwartjens ,  Viktor Marius Boer

IPC: C12N15/81 ,  C12N9/02 ,  C12P19/56 ,  C12N9/88 ,  C12N15/70

Abstract: The present invention relates a variant polypeptide having kaurenoic acid 13-hydroxylase activity, which variant polypeptide comprises an amino acid sequence which, when aligned with a kaurenoic acid 13-hydroxylase comprising the sequence set out in SEQ ID NO: 1, comprises at least one substitution of an amino acid residue corresponding to any of amino acids 72, 85, 108, 127, 129, 141, 172, 195, 196, 197, 199, 226, 236, 291, 302, 361 or 464, said positions being defined with reference to SEQ ID NO: 1 and wherein the variant has one or more modified properties as compared with a reference polypeptide having kaurenoic acid 13-hydroxylase activity. A variant polypeptide of the invention may be used in a recombinant host for the production of steviol or a steviol glycoside.

公开(公告)号：US11725223B2

公开(公告)日：2023-08-15

申请号：US16834756

申请日：2020-03-30

Applicant: DSM IP ASSETS B.V.

Inventor： Viktor Marius Boer ,  Erwin Suir

IPC: C12P19/56 ,  C12N9/88 ,  C12N9/02 ,  C12N9/16 ,  C12N15/52 ,  C12N9/90 ,  C07K14/39 ,  C12N9/10 ,  C12N9/24 ,  A23L27/30 ,  A23L2/60 ,  C07K14/395 ,  C12P15/00 ,  C07H15/256

CPC classification number: C12P19/56 ,  A23L2/60 ,  A23L27/36 ,  C07H15/256 ,  C07K14/39 ,  C07K14/395 ,  C12N9/0036 ,  C12N9/0042 ,  C12N9/0073 ,  C12N9/0083 ,  C12N9/1051 ,  C12N9/16 ,  C12N9/2405 ,  C12N9/88 ,  C12N9/90 ,  C12N15/52 ,  C12P15/00 ,  A23V2002/00

Abstract: The present invention relates to a recombinant microorganism comprising 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, whereby expression of the nucleotide sequence(s) confer(s) on the microorganism the ability to produce at least steviol.

公开(公告)号：US11459548B2

公开(公告)日：2022-10-04

申请号：US17159962

申请日：2021-01-27

Applicant: DSM IP ASSETS B.V.

Inventor： Viktor Marius Boer ,  Johannes Gustaaf Ernst Van Leeuwen ,  Priscilla Zwartjens ,  Catharina Petronella Antonia Maria Kolen

IPC: C12N1/14 ,  C12N1/16 ,  C12N1/20 ,  C12N15/52 ,  C12N15/63 ,  C12P5/00 ,  C12P19/56 ,  C12N9/10 ,  A23L27/30 ,  A23L2/60

Abstract: The present invention relates to a recombinant host comprising a recombinant nucleic acid sequence encoding a polypeptide having at least about: a. 85% identity to the amino acid sequence set forth in SEQ ID NO: 1; b. 85% identity to the amino acid sequence set forth in SEQ ID NO: 3; c. 85% identity to the amino acid sequence set forth in SEQ ID NO: 6; d. 85% identity to the amino acid sequence set forth in SEQ ID NO: 9; e. 85% identity to the amino acid sequence set forth in SEQ ID NO: 11; f. 85% identity to the amino acid sequence set forth in SEQ ID NO: 14; g. 85% identity to the amino acid sequence set forth in SEQ ID NO: 17; h. 85% identity to the amino acid sequence set forth in SEQ ID NO: 20; i. 85% identity to the amino acid sequence set forth in SEQ ID NO: 22; or j. 85% identity to the amino acid sequence set forth in SEQ ID NO: 25.

公开(公告)号：US11178894B2

公开(公告)日：2021-11-23

申请号：US15751674

申请日：2016-08-15

Applicant: DSM IP ASSETS B.V.

Inventor： Viktor Marius Boer ,  Priscilla Zwartjens ,  Eric Van Den Berg

IPC: C07K14/39 ,  C12P19/56 ,  A23L27/30 ,  A23L2/60 ,  C12P15/00 ,  C07K14/395 ,  C07K14/40 ,  C12N1/16 ,  C12N9/24

Abstract: A recombinant host capable of producing a steviol glycoside which overexpresses a polypeptide which mediates steviol glycoside transport and which polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 29 or an amino acid sequence having at least about 50% sequence identity thereto. A recombinant host capable of producing a steviol glycoside which has been modified, preferably in its genome, to result in a deficiency in the production of a polypeptide which mediates steviol glycoside transport and which polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 29 or an amino acid sequence having at least about 50% sequence identity thereto.

公开(公告)号：US10947515B2

公开(公告)日：2021-03-16

申请号：US16777427

申请日：2020-01-30

Applicant: DSM IP ASSETS B.V.

Inventor： Viktor Marius Boer ,  Johannes Gustaaf Emst Van Leeuwen ,  Priscilla Zwartjens ,  Catharina Petronella Antonia Maria Kolen

IPC: C12N1/14 ,  C12N1/16 ,  C12P5/00 ,  C12P19/56 ,  C12N9/10 ,  A23L27/30 ,  A23L2/60 ,  C12N1/20

Abstract: The present invention relates to a recombinant host comprising a recombinant nucleic acid sequence encoding a polypeptide having at least about: a. 85% identity to the amino acid sequence set forth in SEQ ID NO: 1; b. 85% identity to the amino acid sequence set forth in SEQ ID NO: 3; c. 85% identity to the amino acid sequence set forth in SEQ ID NO: 6; d. 85% identity to the amino acid sequence set forth in SEQ ID NO: 9; e. 85% identity to the amino acid sequence set forth in SEQ ID NO: 11; f. 85% identity to the amino acid sequence set forth in SEQ ID NO: 14; g. 85% identity to the amino acid sequence set forth in SEQ ID NO: 17; h. 85% identity to the amino acid sequence set forth in SEQ ID NO: 20; i. 85% identity to the amino acid sequence set forth in SEQ ID NO: 22; or j. 85% identity to the amino acid sequence set forth in SEQ ID NO: 25.

公开(公告)号：US10273519B2

公开(公告)日：2019-04-30

申请号：US14906497

申请日：2014-07-23

Applicant: DSM IP ASSETS B.V.

Inventor： Nicolette Jasmijn Broers ,  Viktor Marius Boer ,  Adam G. Lawrence

IPC: C12P19/56 ,  A23L2/60 ,  C12N9/00 ,  C12N15/52 ,  C12N9/88 ,  C12N9/90 ,  C12N15/00 ,  C07C62/32 ,  C07H15/256 ,  C12N9/02 ,  C12N9/10 ,  C12P15/00 ,  A23L27/30

Abstract: 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.