Bacterial strain producing 2,3-butanediol and other metabolites

Patent metadata

Field	Value
Publication number	US-11111474-B2
Application number	US-201716316961-A
Country	US
Kind code	B2
Filing date	Jul 18, 2017
Priority date	Jul 19, 2016
Publication date	Sep 7, 2021
Grant date	Sep 7, 2021

Abstract

The present invention provides a modified strain of the bacterium Lactococcus lactis obtainable by a method that comprises a step of fusion of two protoplasts from two Lactococcus lactis parental strains which, compared to the wild type strain of Lactococcus lactis from which they derive, show: (a) an increased ability to produce acetoin and/or 2,3-butanediol (2,3-BDO), and (b) a decreased ability to produce lactic acid, when cultured under aerobic conditions, and wherein the modified strain of Lactococcus lactis has an increased ability to produce 2,3-BDO of at least 20 times the amount produced by the wild type strain, an increased ability to produce acetoin of at least 20 times the amount produced by the wild type strain and a decreased ability to produce lactic acid of at least 10 times the amount produced by the wild type strain, when cultured under aerobic conditions. Also provided are methods to produce acetoin and 2,3-BDO.

First claim

The invention claimed is:

1. A Lactococcus lactis strain 43103 deposited at the Spanish Type Culture Collection under accession number CECT 9139, wherein said strain has an increased ability to produce acetoin and 2,3-butanediol and a decreased ability to produce lactic add when cultured under aerobic conditions.

2. A method of production of 2,3-butanediol that comprises the aerobic fermentation of a carbohydrate-containing culture medium by the bacterial strain as defined in claim 1 .

3. The method of 2,3-butanediol production according to claim 2 , comprising the following steps: (a) Pre-culture of the Lactococcus lactis strain 43103 deposited at the Spanish Type Culture Collection under accession number CECT 9139; (b) Inoculation of a carbohydrate-containing culture medium with the pre-culture obtained in step (a); (c) Fermentation of carbohydrates present in the culture medium inoculated in step (b) at 20-40° C.; pH 5.5-6.0 and 5-100% dissolved oxygen concentration; and (d) Separation of cells from the fermented broth resulting from step (c).

4. The method of 2,3-butanediol production according to claim 3 , further comprising a step of purification of the 2,3-butanediol present in the cell-free fermentation broth resulting from step (d).

5. A method of production of acetoin, that comprises the aerobic fermentation of a carbohydrate-containing culture medium by the bacterial strain as defined in claim 1 .

6. The method of acetoin production according to claim 5 , comprising the following steps: (a) Pre-culture of the Lactococcus lactis strain 43103 strain deposited at the Spanish Type Culture Collection under accession number CECT 9139; (b) Inoculation of a carbohydrate-containing culture medium with the pre-culture obtained in step (a); (c) Fermentation of carbohydrates present in the culture medium inoculated in step (b) at 20-40° C.; pH 6.5-7.5 and 30-100% dissolved oxygen concentration; and (d) Separation of cells from the fermented broth resulting from step (c).

7. The method of acetoin production according to claim 6 , further comprising a step of purification of the acetoin present in the cell-free fermentation broth resulting from step (d).

8. A method of production of lactic add that comprises the anaerobic fermentation of a carbohydrate-containing culture medium by the bacterial strain as defined in claim 1 .

Assignees

• Fundacion Tecnalia Res & Innovation

Inventors

• Roncal Martínez Tomás
• Caballero Román Susana
• Díaz De Guereñu Zabarte María Del Mar
• Rincón Arroyo Inés
• Prieto Fernández Soraya
• Ochoa Gómez José Ramón

Classifications

• C12P7/18

  polyhydric · CPC title

• C12P7/56

  Lactic acid · CPC title

• C12N15/01

  Preparation of mutants without inserting foreign genetic material therein; Screening processes therefor · CPC title

• C12N1/20 Primary

  Bacteria; Culture media therefor · CPC title

• C12N1/205 Primary

  Bacterial isolates · CPC title