Ethanol production

Abstract

This invention relates to ethanol production as a product of bacterial fermentation.

1 . A Gram-positive bacterium which has been transformed with a heterologous gene encoding pyruvate decarboxylase or a functional equivalent thereof, but has solely native alcohol dehydrogenase function.  
     
     
         2 . A Gram-positive bacterium according to  claim 1  wherein the bacterium is a  Bacillus sp.    
     
     
         3 . A Gram-positive bacterium according to  claim 1  or  2  wherein the bacterium is a thermophile.  
     
     
         4 . A Gram-positive bacterium according to  claim 2  or  3  wherein the  Bacillus  is selected from  B. stearothermophilus; B. calvodax; B. caldotenax, B. thermoglucosidasius, B. coagulans, B. licheniformis, B. thermodenitrificans,  and  B. caldolyticus.    
     
     
         5 . A Gram-positive bacterium according to  claim 1 ,  2 ,  3  or  4  wherein the gene encoding lactate dehydrogenase expression has been inactivated.  
     
     
         6 . A Gram-positive bacterium according to  claim 5  in which the lactate dehydrogenase gene has been inactivated by homologous recombination.  
     
     
         7 . A Gram-positive bacterium according to any preceding claim in which the heterologous gene is from  Zymomonas sp  or from  Saccharomyces cerevisiae.    
     
     
         8 . A Gram-positive bacterium according to  claim 7  in which the heterologous gene is from  Z. mobilis.    
     
     
         9 . A Gram-positive bacteria according to  claim 7  in which the heterologous gene is pdc 5 from  S. cerevisiae.    
     
     
         10 . A Gram-positive bacterium according to the preceding claim wherein the heterologous gene is incorporated into the chromosome of the bacterium.  
     
     
         11 . A Gram-positive bacterium according to any one of  claims 1  to  9  in which the bacterium has been transformed with a plasmid comprising the heterologous gene.  
     
     
         12 . A Gram-positive bacterium according to  claim 11 , wherein the plasmid is pFC1.  
     
     
         13 . A Gram-positive bacteria according to any one of  claims 1  to  9 , wherein the heterologous gene is operatively linked to the lactate dehydrogenase promoter from  Bacillus  strain LN (NCIMB accession number 41038).  
     
     
         14 . Strains LN (NCIMB accession number 41038); LN-T (E31, E32); TN NCIMB accession number 41039); TN-P1; TN-P3; LN-S (J8) (NCIMB accession number 41040); LN-D (NCIMB accession number 41041); LN-D11 and LN-P1.  
     
     
         15 . A recombinant, sporulation deficient, thermophilic  Bacillus  which grows at greater than 50° C.  
     
     
         16 . A recombinant sporulation deficient, thermophilic  Bacillus  which grows at greater than 50° C. and which is not  B. licheniformis.    
     
     
         17 . A method of producing ethanol comprising culturing a bacterium or strain according to any one of  claims 1  to  13  under suitable conditions.  
     
     
         18 . A method according to  claim 17  in which the method is operated at a temperature between 40-75° C.  
     
     
         19 . A method according to  claim 18  operated at a temperature of 52-65° C.  
     
     
         20 . A method according to  claim 18  operated at a temperature of 60-65° C.  
     
     
         21 . A method of producing L-lactic acid comprising culturing strain LN under suitable conditions.  
     
     
         22 . A nucleic acid molecule comprising the lactate dehydrogenase promoter region of strain LN (NCIMB accession number 41038).  
     
     
         23 . The nucleic acid molecule of  claim 22 , wherein the nucleic acid molecule comprises the nucleic acid sequence shown in FIG. 8.  
     
     
         24 . Plasmid pFC1.  
     
     
         25 . Plasmid pFC1-PDC1.