Method of screening gene for 1,4-bdo production

Patent metadata

Field	Value
Publication number	US-2016208292-A1
Application number	US-201414913767-A
Country	US
Kind code	A1
Filing date	Aug 22, 2014
Priority date	Aug 23, 2013
Publication date	Jul 21, 2016
Grant date	—

Abstract

Provided is a screening method of discovering genes capable of increasing 1,4-BDO production on the basis of proteomics data. Over-expression of proteins screened by the method, NCgl0630 (citrate synthase) and NCgl2145 (hyperthetical protein), increase 1,4-BDO productivity. The method may lead to screening of a protein associated with 1,4-BDO productivity, thereby increasing 1,4-BDO productivity, and thus, the method may be recognized as being industrially applicable.

First claim

1 . A method of screening for a protein positively involved in 1,4-BDO production, comprising:
culturing a wild-type microorganism incapable of producing 1,4-BDO and a mutant microorganism capable of producing 1,4-BDO in a culture medium with or without 1,4-BDO; analyzing protein expression in the culture solution of the microorganisms; and selecting a protein for which expression is higher in the mutant microorganism capable of producing 1,4-BDO than that in the wild type microorganism as a protein positively involved in 1,4-BDO production.

2 . The method of claim 1 , wherein the culturing comprises culturing the wild type microorganism incapable of producing 1,4-BDO and the mutant microorganism capable of producing 1,4-BDO in culture mediums including 1,4-BDO of one or more concentration.

3 . The method of claim 1 , wherein the microorganism is Corynebacterium glutamicum.

4 . The method of claim 1 , wherein the mutant microorganism does not have lactate dehydrogenase activitym.

5 . The method of claim 1 , wherein the mutant microorganism comprises genes encoding coenzyme A-dependent succinate semialdehyde dehydrogenase, 4-hydroxybutyrate dehydrogenase, 4-hydroxybutyryl CoA: acetyl-CoA transferase, and alcohol dehydrogenase.

6 . The method of claim 1 , wherein the 1,4-BDO is added to the culture medium in an exponential phase of growth of the microorganism.

7 - 11 . (canceled)

12 . A microorganism capable of producing 1,4-BDO, wherein a nucleic acid sequence encoding an amino acid sequence having at least 95% of sequence identity with of SEQ ID NO:3 is additionally introduced.

13 . The microorganism of claim 12 , wherein the nucleic acid sequence encoding an amino acid sequence of SEQ ID NO:3 is SEQ ID NO:4.

14 . The microorganism of claim 12 , wherein the microorganism is Corynebacterium glutamicum.

15 . The microorganism of claim 12 , wherein a nucleic acid sequence encoding citrate synthase is additionally introduced.

16 . The microorganism of claim 12 , wherein the microorganism overexpresses the nucleic acid sequence compared to a parent microorganism not having the additionally introduced nucleic acid sequence.

17 . (canceled)

18 . A method of producing 1,4-BDO comprising:
culturing the microorganism of claim 12 ; and obtaining 1,4-BDO from culture solution.

Assignees

• Samsung Electronics Co Ltd

Inventors

• Cho Hwayoung
• Park Jinhwan
• Jung Yukyung
• Park Jaechan

Classifications

• C12P7/18 Primary

  polyhydric · CPC title

• C12N9/1025

  Acyltransferases (2.3) · CPC title

• C12N9/0006

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

• G01N2333/91045

  Acyltransferases (2.3) · CPC title

• C12Y101/01027

  L-Lactate dehydrogenase (1.1.1.27) · CPC title