Use of thiamine and nicotine adenine dinucleotide for butanol production

Abstract

The invention relates generally to the field of industrial microbiology and alcohol production. More specifically, the invention relates to the use of thiamine, biosynthetic precursors of thiamine, nicotinic acid, nicotinamid, nicotinic acid riboside, nicotinamid riboside, or other biosynthetic precursors of nicotine adenine dinucleotide (NAD) to improve butanol production. Butanol production can be improved by providing sufficient amounts of thiamine, biosynthetic precursors of thiamine, nicotinic acid, nicotinamid, nicotinic acid riboside, nicotinamid riboside, or other biosynthetic precursors of nicotine adenine dinucleotide (NAD) in the production media.

1 - 15 . (canceled) 
     
     
         16 . A commercial method for producing a fermentation product comprising:
 a. providing a production culture comprising recombinant cells capable of producing butanol and a production media comprising
 1. nicotinic acid, nicotinamid, or a biosynthetic precursor of NAD; 
 2. optionally thiamine or a biosynthetic precursor thereof; and 
 3. a production feed derived from biomass comprising a fermentable carbon source; 
   
       wherein the production media contains less than about 1 g/L multi-component media additives; and
 b. contacting the production culture with the production media in a fermentation vessel to form a production broth under conditions whereby a fermentation product is produced. 
 
     
     
         17 . The method of  claim 16 , wherein the production media is substantially free of multi-component media additives. 
     
     
         18 . The method of  claim 16 , wherein the production broth contains at least about 5 mg/L nicotinic acid, nicotinamid, or a biosynthetic precursor of NAD. 
     
     
         19 . The method of  claim 16 , wherein the nicotinic acid, nicotinamid, or a biosynthetic precursor of NAD thereof is present in an amount sufficient to provide at least about 30% of the rate, titer, or specific productivity observed in media that contains about 10 g/L yeast extract. 
     
     
         20 . The method of  claim 16 , wherein the recombinant cells are yeast. 
     
     
         21 . The method of  claim 20 , wherein the yeast is crabtree positive or crabtree negative. 
     
     
         22 . The method of  claim 16 , wherein the recombinant cell comprises a modification of an endogenous gene encoding a thiamine-phyrophosphate-dependent (TPP) gene. 
     
     
         23 . The method of  claim 22 , wherein the TPP-dependent gene is a pyruvate decarboxylase gene. 
     
     
         24 . The method of  claim 16 , wherein the recombinant cell comprises a butanol biosynthetic pathway. 
     
     
         25 . The method of  claim 24 , wherein the butanol biosynthetic pathway is comprised of at least one heterologous polynucleotide encoding a polypeptide that catalyzes a substrate to product conversion of the pathway. 
     
     
         26 . The method of  claim 24 , wherein the butanol biosynthetic pathway is an isobutanol biosynthetic pathway. 
     
     
         27 . The method of  claim 26 , wherein the isobutanol biosynthetic pathway comprises the following substrate to product conversions:
 a) pyruvate to acetolactate;   b) acetolactate to 2,3-dihydroxyisovalerate;   c) 2,3-dihydroxyisovalerate to α-ketoisovalerate;   d) α-ketoisovalerate to isobutyraldehyde; and   e) isobutyraldehyde to isobutanol.   
     
     
         28 . The production broth of  claim 16 , wherein the broth comprises butanol. 
     
     
         29 . A butanol fermentation production media comprising;
 1. nicotinic acid, nicotinamid, or a biosynthetic precursor of NAD;   2. optionally thiamine or a biosynthetic precursor thereof; and   3. a production feed derived from biomass comprising a fermentable carbon source; and   
       wherein the production media comprises less than 1 g/L of multi-component media additives. 
     
     
         30 . The production media of  claim 29 , consisting essentially of greater than about 10 mg/L nicotinic acid, nicotinamid, or a biosynthetic precursor of NAD; optionally, greater than about 10 mg/L thiamine or a biosynthetic precursor thereof; and liquefied biomass comprising a fermentable carbon substrate. 
     
     
         31 . A recombinant host cell comprising at least one DNA molecule encoding a polypeptide having aspartate oxidase or aspartate dehydrogenase activity. 
     
     
         32 . A recombinant host cell comprising at least one DNA molecule encoding a polypeptide having quinolinate synthase activity.