Method of screening gene for 1,4-bdo production

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.

1 . A method of screening for a protein positively involved in 1,4-BDO production, comprising:
 culturing a microorganism that produces 1,4-BDO in a culture medium comprising 1,4-BDO and in a culture medium not comprising 1,4-BDO;   analyzing protein expression in the microorganisms; and   selecting a protein showing increased expression in the microorganism cultured with 1,4-BDO as compared to the microorganism cultured without 1,4-BDO   as a protein positively involved in 1,4-BDO production.   
     
     
         2 . The method of  claim 1 , wherein the microorganism that produces 1,4-BDO is a mutant microorganism, and the method further comprises culturing a wild type microorganism incapable of producing 1,4-BDO in a culture medium comprising 1,4-BDO and in a culture medium not comprising 1,4-BDO a protein showing (a) increased expression in the microorganism cultured with 1,4-BDO as compared to the microorganism cultured without 1,4-BDO, and (b) increased expression in the mutant microorganism capable of producing 1,4-BDO than in the wild type microorganism. 
     
     
         3 . The method of  claim 1 , wherein the microorganism is Corynebacterium glutamicum. 
     
     
         4 . The method of  claim 1 , wherein the microorganism does not have lactate dehydrogenase activity. 
     
     
         5 . The method of  claim 1 , wherein the 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 during an exponential growth phase of the microorganism. 
     
     
         7 - 18 . (canceled) 
     
     
         19 . The method of  claim 2 , wherein the mutant microorganism is Corynebacterium glutamicum modified to produce 1,4-BDO, and the wild-type microorganism is Corynebacterium glutamicum.