公开(公告)号：US20210040491A1

公开(公告)日：2021-02-11

申请号：US16938483

申请日：2020-07-24

Applicant: DSM IP ASSETS B.V.

Inventor： Johannes Gustaaf Ernst VAN LEEUWEN ,  Priscilla Zwartjens ,  Viktor Marius Boer

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

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：US10604743B2

公开(公告)日：2020-03-31

申请号：US15558133

申请日：2016-03-16

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 ,  C12N9/10 ,  C12P5/00 ,  C12P19/56 ,  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：US12091436B2

公开(公告)日：2024-09-17

申请号：US18167119

申请日：2023-02-10

Applicant: DSM IP ASSETS B.V.

Inventor： Johannes Gustaaf Ernst Van Leeuwen ,  Viktor Marius Boer ,  Priscilla Zwartjens ,  Jos Van Vugt ,  René Marcel De Jong

IPC: C07K14/39 ,  C12N9/02 ,  C12P19/56

CPC classification number: C07K14/39 ,  C12N9/0042 ,  C12P19/56 ,  C12Y106/02004

Abstract: The disclosure provides a recombinant cell capable of producing a steviol glycoside, wherein the cell comprises a nucleic acid coding for a variant of a parent polypeptide, wherein the variant has steviol glycoside transport mediating activity, wherein the variant comprises an amino acid sequence which, when aligned with the amino acid sequence of the parent polypeptide, comprises at least one modification of the amino acid residue corresponding to any of the amino acids in the amino acid sequence of the parent polypeptide, wherein the variant has an improved ability to produce rebaudioside M and optionally other steviol glycosides extracellularly if compared with the parent polypeptide when measured under the same conditions.

公开(公告)号：US11925193B2

公开(公告)日：2024-03-12

申请号：US17512368

申请日：2021-10-27

Applicant: DSM IP ASSETS B.V.

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

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

CPC classification number: A23L27/36 ,  A23L2/60 ,  C07K14/39 ,  C07K14/395 ,  C07K14/40 ,  C12N1/16 ,  C12N9/2402 ,  C12P15/00 ,  C12P19/56

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：US11297862B2

公开(公告)日：2022-04-12

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：US11104886B2

公开(公告)日：2021-08-31

申请号：US16467087

申请日：2017-12-04

Applicant: DSM IP Assets B.V.

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

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

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：US20200283816A1

公开(公告)日：2020-09-10

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: 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.