Use of thiamine and nicotine adenine dinucleotide for butanol production

摘要

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.

主权项

1. A commercial method for producing isobutanol comprising:
a. providing a production culture comprising recombinant cells, wherein the recombinant cells comprise an engineered isobutanol biosynthetic pathway, wherein the engineered isobutanol biosynthetic pathway comprises heterologous polynucleotides encoding polypeptides that catalyze the following substrate to product conversions of the pathway:
a) pyruvate to acetolactate, catalyzed by an acetolactate synthase;b) acetolactate to 2,3-dihydroxyisovalerate, catalyzed by an acetohydroxy acid reductoisomerase;c) 2,3-dihydroxyisovalerate to a-ketoisovalerate, catalyzed by a acetohydroxy acid dehydratase;d) a-ketoisovalerate to isobutyraldehyde, catalyzed by a branched-chain keto acid decarboxylase; ande) isobutyraldehyde to isobutanol, catalyzed by a branched-chain alcohol dehydrogenase,wherein the recombinant cells comprise at least one genetic modification that eliminates pyruvate decarboxylase activity,and wherein the recombinant cells are capable of producing isobutanol, and production media comprising
1. thiamine or a biosynthetic precursor thereof;2. optionally nicotinic acid, nicotinamid, or a biosynthetic precursor of NAD; and3. a production feed derived from biomass comprising a fermentable carbon source;wherein the production media contains less than 1 g/L of 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 isobutanol is produced.