Host cells and their use for producing ribitol and further monosaccharides

Patent metadata

Field	Value
Publication number	US-12522848-B2
Application number	US-202017779561-A
Country	US
Kind code	B2
Filing date	Dec 17, 2020
Priority date	Dec 18, 2019
Publication date	Jan 13, 2026
Grant date	Jan 13, 2026

Abstract

The present invention relates to host cells and their use wherein the host cells are capable of producing D-ribulose and incapable of or have a reduced capability of converting D-ribulose to a molecule other than ribitol, wherein the host cells comprise a heterologous nucleic acid sequence encoding a polypeptide capable of converting D-ribulose to ribitol with a cofactor preference for NADPH.

First claim

What is claimed is:

1 . A host cell that is capable of producing D-ribulose, wherein the host cell comprises one or more heterologous nucleic acids encoding a polypeptide capable of converting D-ribulose to ribitol selected from:
(a) a nucleic acid encoding a polypeptide capable of converting D-ribulose to ribitol comprising an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 7, wherein said polypeptide has a serine and an arginine at the positions corresponding to positions 45 and 46 of the polypeptide of SEQ ID NO: 7, respectively; (b) a nucleic acid encoding a polypeptide capable of converting D-ribulose to ribitol comprising an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 9, wherein said polypeptide has a serine and an arginine at the positions corresponding to positions 45 and 46 of the polypeptide of SEQ ID NO: 9, respectively; and (c) a nucleic acid encoding a polypeptide capable of converting D-ribulose to ribitol comprising an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 11, wherein said polypeptide has a serine and an arginine at the positions corresponding to positions 38 and 39 of the polypeptide of SEQ ID NO: 11, respectively;
wherein the host cell comprises an endogenous gene encoding a polypeptide that is capable of converting D-ribulose to D-arabitol, wherein said endogenous gene is inactivated through deletion of the gene, and wherein the host cell is a yeast cell.

2 . The host cell of claim 1 , wherein the host cell is capable of producing D-ribulose from D-glucose.

3 . The host cell of claim 1 , wherein the host cell comprises a heterologous nucleic acid encoding a polypeptide capable of converting D-ribulose to ribitol comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 7, and wherein said polypeptide has a serine and an arginine at the positions corresponding to positions 45 and 46 of the polypeptide of SEQ ID NO: 7, respectively.

4 . The host cell of claim 1 , wherein the host cell comprises a heterologous nucleic acid encoding a polypeptide capable of converting D-ribulose to ribitol comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 9, and wherein said polypeptide has a serine and an arginine at the positions corresponding to positions 45 and 46 of the polypeptide of SEQ ID NO: 9, respectively.

5 . The host cell of claim 1 , wherein the host cell comprises a heterologous nucleic acid encoding a polypeptide capable of converting D-ribulose to ribitol comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 11, and wherein said polypeptide has a serine and an arginine at the positions corresponding to positions 38 and 39 of the polypeptide of SEQ ID NO: 11, respectively.

6 . The host cell of claim 1 , wherein the yeast cell is Pichia ohmeri.

7 . The host cell of claim 1 , wherein the host cell comprises a heterologous nucleic acid encoding a polypeptide capable of converting D-ribulose to ribitol comprising SEQ ID NO 7.

8 . The host cell of claim 1 , wherein the host cell comprises a heterologous nucleic acid encoding a polypeptide capable of converting D-ribulose to ribitol comprising SEQ ID NO 9.

9 . The host cell of claim 1 , wherein the host cell comprises a heterologous nucleic acid encoding a polypeptide capable of converting D-ribulose to ribitol comprising SEQ ID NO 11.

10 . The host cell of claim 1 , wherein the host cell comprises more than one of the heterologous nucleic acids encoding a polypeptide capable of converting D-ribulose to ribitol.

11 . The host cell of claim 10 , wherein the host cell comprises (a) a heterologous nucleic acid encoding a polypeptide capable of converting D-ribulose to ribitol comprising an amino acid sequence having at least 85% sequence identity to SEQ ID NO:7, wherein said polypeptide has a serine and an arginine at the positions corresponding to positions 45 and 46 of the polypeptide of SEQ ID NO: 7, respectively, and (b) a heterologous nucleic acid encoding a polypeptide capable of converting D-ribulose to ribitol comprising an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 9, wherein said polypeptide has a serine and an arginine at the positions corresponding to positions 45 and 46 of the polypeptide of SEQ ID NO: 9, respectively.

12 . The host cell of claim 1 , wherein the host cell comprises a heterologous nucleic acid encoding a polypeptide capable of converting D-ribulose to ribitol comprising an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 7, and wherein said polypeptide has a serine and an arginine at the positions corresponding to positions 45 and 46 of the polypeptide of SEQ ID NO: 7, respectively.

13 . The host cell of claim 1 , wherein the host cell comprises a heterologous nucleic acid encoding a polypeptide capable of converting D-ribulose to ribitol comprising an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 9, and wherein said polypeptide has a serine and an arginine at the positions corresponding to positions 45 and 46 of the polypeptide of SEQ ID NO: 9, respectively.

14 . The host cell of claim 1 , wherein the host cell comprises a heterologous nucleic acid encoding a polypeptide capable of converting D-ribulose to ribitol comprising an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 11, and wherein said polypeptide has a serine and an arginine at the positions corresponding to positions 38 and 39 of the polypeptide of SEQ ID NO: 11, respectively.

15 . A method for producing ribitol comprising culturing the host cell of claim 1 under cell culture conditions, wherein ribitol is produced by the host cell.

16 . The method of claim 15 , further comprising isolating the produced ribitol.

Assignees

• Danstar Ferment Ag

Inventors

• Schäfer Astrid
• Chang Yiming
• Malca Sumire Honda

Classifications

• C12N15/80

  for fungi · CPC title

• C12N9/0006 Primary

  acting on CH-OH groups as donors (1.1) · CPC title

• C12N1/16

  Yeasts; Culture media therefor · CPC title

• C12P7/18 Primary

  polyhydric · CPC title

• C12N9/0004

  Oxidoreductases (1.) · CPC title