Improved production of riboflavin

Patent metadata

Field	Value
Publication number	US-2022010291-A1
Application number	US-201917293800-A
Country	US
Kind code	A1
Filing date	Nov 11, 2019
Priority date	Nov 15, 2018
Publication date	Jan 13, 2022
Grant date	—

Abstract

The present invention is related to an improved process for production of vitamin B2 using a genetically engineered host cell comprising a heterologous enzyme with pyridoxal phosphatase activity. Using said modified host cell, the yield of riboflavin production could be increased by at least about 5%.

First claim

1 . A riboflavin-producing host cell comprising riboflavin biosynthetic genes (rib operon) and a heterologous enzyme with pyridoxal phosphatase [EC 3.1.3.74] activity.

2 . The riboflavin-producing host cell of claim 1 comprising the riboflavin biosynthetic genes from Bacillus.

3 . The riboflavin-producing host cell of claim 1 comprising a polypeptide with at least about 70% identity to SEQ ID NO:2.

4 . The riboflavin-producing host cell according to claim 3 , expressing a polynucleotide encoding a polypeptide with at least 70% identity to SEQ ID NO:2.

5 . The riboflavin-producing host cell according to claim 1 , wherein the heterologous enzyme is selected from bacteria, preferably Rhizobia , more preferably Sinorhizobium.

6 . The riboflavin-producing host cell according to claim 1 , wherein the activity of the endogenous ribC gene has been reduced.

7 . The riboflavin-producing host cell according to claim 1 producing wherein the yield of riboflavin is increased by at least about 5% from a given carbon source compared to production of riboflavin using a host cell not expressing a polynucleotide encoding a polypeptide with at least 70% identity to SEQ ID NO:2.

8 . The riboflavin-producing host cell according to claim 1 , which is selected from Bacillus, Corynebacterium, Streptococcus, Clostridium, Lactococcus or Streptomyces , preferably selected from the group consisting of Bacillus subtilis, Bacillus cereus, Bacillus amyloliquefaciens, Bacillus stearothermophilus, Bacillus halodurans, Bacillus licheniformis, Streptococcus aureus, Streptococcus pneumoniae, Clostridium acetobutylicum, Clostridium difficile, Lactococcus lactis, Streptomyces coelicolor, Corynebacterium diphteriae and Corynebacterium glutamicum.

9 . Process for production of riboflavin, wherein the riboflavin-producing host cell according to claim 1 is incubated in an aqueous medium under conditions that allow the production of riboflavin from a given substrate.

10 . The process according to claim 9 , wherein the yield of riboflavin is increased by at least 5% compared to a process wherein the host cell does not express a polynucleotide encoding a polypeptide with at least 70% identity to SEQ ID NO:2.

11 . A process according to claim 9 comprising the steps of:
(a) providing a riboflavin-producing host cell according to any one of claims 1 to 7 ,
(b) incubating said host cell in an aqueous medium under conditions that allow the production of riboflavin from a given substrate, and optionally
(c) isolating and purifying the riboflavin from the cultivation medium.

12 . Use of a polypeptide with at least 70% identity to SEQ ID NO:2 or of a riboflavin-producing host cell according to claim 1 in a process for production of riboflavin.

13 . A riboflavin-producing host cell comprising riboflavin biosynthetic genes (rib operon) and a heterologous enzyme having pyridoxal phosphatase activity [EC 3.1.3.74] used for a process for production of riboflavin.

Assignees

• Dsm Ip Assets Bv

Inventors

• Farrell Christopher Mark
• Potot Sébastien Eric
• Prágai Zoltán
• Anaya Michael Patrick

Classifications

• C12N9/16 Primary

  acting on ester bonds (3.1) · CPC title

• C12N15/52

  Genes encoding for enzymes or proenzymes · CPC title

• C12P17/165

  Heterorings having nitrogen atoms as the only ring heteroatoms · CPC title

• C12Y301/03074

  Pyridoxal phosphatase (3.1.3.74) · CPC title

• C12P25/00 Primary

  Preparation of compounds containing alloxazine or isoalloxazine nucleus, e.g. riboflavin · CPC title