Steviol glycoside transport

Patent metadata

Field	Value
Publication number	US-11297862-B2
Application number	US-201615751674-A
Country	US
Kind code	B2
Filing date	Aug 15, 2016
Priority date	Aug 13, 2015
Publication date	Apr 12, 2022
Grant date	Apr 12, 2022

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.

First claim

The invention claimed is:

1. A recombinant cell capable of producing a steviol glycoside which overexpresses a nucleic acid encoding a polypeptide comprising the amino acid sequence set forth in SEQ ID NO:29 or an amino acid sequence having at least 98% sequence identity to the amino acid sequence set forth in SEQ ID NO:29.

2. The recombinant cell according to claim 1 , which further comprises one or more recombinant 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.

3. The recombinant cell according to claim 1 , wherein the host belongs to one of the genera Saccharomyces, Aspergillus, Pichia, Kluyveromyces, Candida, Hansenula, Humicola, Issatchenkia, Trichosporon, Brettanomyces, Pachysolen, Yarrowia , Yamadazyma, or Escherichia.

4. The recombinant host according to claim 3 , wherein the recombinant host is a Saccharomyces cerevisiae cell, a Yarrowia lipolytica cell, a Candida krusei cell, or an Escherichia coli cell.

5. The recombinant cell according to claim 1 , wherein the polypeptide comprises an amino acid sequence having at least 99% sequence identity to SEQ ID NO:29.

6. The recombinant cell according to claim 1 , wherein the polypeptide mediates steviol glycoside transport and wherein the amount of total produced steviol glycoside outside the cell, as compared to inside the cell, is higher as compared to a corresponding cell which does not overexpress the nucleic acid.

7. A process for the preparation of a steviol glycoside which comprises fermenting the recombinant cell according to claim 1 in a suitable fermentation medium and, optionally, recovering the steviol glycoside.

8. The process according to claim 7 for the preparation of a steviol glycoside, wherein the process is carried out on an industrial scale.

Assignees

• Dsm Ip Assets Bv

Inventors

• Boer Viktor Marius
• Zwartjens Priscilla
• Van Den Berg Eric

Classifications

• C12Y505/01013

  Ent-copalyl diphosphate synthase (5.5.1.13) · CPC title

• C12Y402/03019

  Ent-kaurene synthase (4.2.3.19) · CPC title

• C12Y114/13078

  Ent-kaurene oxidase (1.14.13.78) · CPC title

• C12Y106/02004

  NADPH-hemoprotein reductase (1.6.2.4), i.e. NADP-cytochrome P450-reductase · CPC title

• C12Y101/01088

  Hydroxymethylglutaryl-CoA reductase (1.1.1.88) · CPC title