Production of riboflavin

Patent metadata

Field	Value
Publication number	US-12365885-B2
Application number	US-201917293800-A
Country	US
Kind code	B2
Filing date	Nov 11, 2019
Priority date	Nov 15, 2018
Publication date	Jul 22, 2025
Grant date	Jul 22, 2025

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

The invention claimed is:

1. A riboflavin-producing Bacillus subtilis host cell expressing a polynucleotide encoding a heterologous enzyme with pyridoxal phosphatase activity, wherein the heterologous enzyme comprises a polypeptide with at least 95% sequence identity to the polypeptide of SEQ ID NO:2, and wherein the host cell comprises a mutation in an endogenous ribC gene that disrupts the expression of the endogenous ribC gene.

2. The riboflavin-producing host cell according to claim 1 , wherein the heterologous enzyme is a Sinorhizobium meliloti enzyme.

3. The riboflavin-producing host cell according to claim 1 , wherein the yield of riboflavin is increased by at least 5% from a given carbon source compared to the yield of riboflavin using the corresponding host cell not expressing the polynucleotide encoding the heterologous enzyme.

4. The riboflavin-producing host cell according to claim 1 , wherein the heterologous enzyme comprises a polypeptide with at least 98% sequence identity to the polypeptide of SEQ ID NO: 2.

5. A process for production of riboflavin, comprising
(a) incubating a riboflavin-producing microbial host cell in an aqueous medium under conditions that allow the production of riboflavin from a given substrate; and
(b) isolating the riboflavin from the cultivation medium,
wherein the riboflavin-producing host cell expresses a polynucleotide encoding a heterologous enzyme with pyridoxal phosphatase activity, wherein the heterologous enzyme comprises a polypeptide with at least 95% sequence identity to the polypeptide of SEQ ID NO:2.

6. The process according to claim 5 , wherein the yield of riboflavin is increased by at least 5% compared to a process using the corresponding host cell that does not express the heterologous enzyme.

7. The process according to claim 5 further comprising the step of isolating and purifying the riboflavin from the cultivation medium.

8. The process according to claim 5 , wherein the host cell is 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. The process according to claim 5 , wherein the host cell is Bacillus subtilis.

10. The process according to claim 9 , wherein the host cell comprises a mutation in an endogenous ribC gene that disrupts the expression of the endogenous ribC gene.

11. The process according to claim 5 , wherein the heterologous enzyme is a Sinorhizobium meliloti enzyme.

12. The process according to claim 5 , wherein the heterologous enzyme comprises a polypeptide with at least 98% sequence identity to the polypeptide of SEQ ID NO: 2.

Assignees

• Dsm Ip Assets Bv

Inventors

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

Classifications

• C12Y301/03074

  Pyridoxal phosphatase (3.1.3.74) · CPC title

• C12P17/165

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

• C12N15/52

  Genes encoding for enzymes or proenzymes · CPC title

• C12N9/16 Primary

  acting on ester bonds (3.1) · CPC title

• C12P25/00 Primary

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