In situ expression of lipase for enzymatic production of alcohol esters during fermentation

Abstract

Disclosed herein are methods of producing alcohol esters during a fermentation by providing alcohol-producing microorganisms which further comprise an engineered polynucleotide encoding a polypeptide having lipase activity.

What is claimed: 
     
         1 - 23 . (canceled) 
     
     
         24 . A recombinant host cell comprising:
 (a) an engineered alcohol production pathway; and   (b) an engineered polynucleotide encoding a polypeptide having lipase activity.   
     
     
         25 . The recombinant host cell of  claim 24  wherein the polypeptide having lipase activity comprises a sequence having at least about 70% identity to SEQ ID NO:2, 4, 6, 256, 47, 49, 51, 53, 55, 241, 242, 243, 244, 245, 246, 247, 248, 272, or 274 or a fragment thereof having lipase activity. 
     
     
         26 . The recombinant host cell of  claim 24  wherein the polypeptide having lipase activity further comprises a sequence having at least about 70% identity to anyone of SEQ ID NOs: 249, 250, 251, 252, 253 or a fragment thereof having lipase activity. 
     
     
         27 . The recombinant host cell of  claim 26  wherein the polypeptide having lipase activity does not contain a glycosylation motif. 
     
     
         28 . The recombinant host cell of  claim 24  wherein the polypeptide having lipase activity is not glycosylated. 
     
     
         29 . The recombinant host cell of  claim 24  wherein the engineered polynucleotide encoding a polypeptide having lipase activity comprises a sequence having at least about 70% identity to SEQ ID NO: 1, 3, 5, 7, 8, 9, 46, 48, 50, 52, 54, 255, 271 or 273. 
     
     
         30 . A recombinant host cell comprising:
 (a) an alcohol production pathway; and   (b) an engineered polynucleotide encoding a polypeptide having lipase activity wherein the polypeptide having lipase activity comprises a sequence having at least about 70% identity to SEQ ID NO: 2, 4, 6, 256, 47, 49, 51, 53, 55, 241, 242, 243, 244, 245, 246, 247, 248, 272, or 274 or an active fragment thereof.   
     
     
         31 . The recombinant host cell of  claim 30  wherein the polypeptide having lipase activity further comprises a sequence having at least about 70% identity to any one of SEQ ID NOs: 249, 250, 251, 252, 253 or a fragment thereof. 
     
     
         32 . The recombinant host cell of  claim 24  wherein the alcohol production pathway is a butanol production pathway 
     
     
         33 . The recombinant host cell of  claim 32  wherein the butanol production pathway is an isobutanol production pathway 
     
     
         34 . The recombinant host cell of  claim 24  wherein the host cell further comprises reduced or eliminated pyruvate decarboxylase activity. 
     
     
         35 - 46 . (canceled) 
     
     
         47 . A fermentation medium comprising an alcohol-producing microorganism comprising a butanol biosynthetic pathway and further comprising an engineered polynucleotide encoding a polypeptide having lipase activity which is expressed and secreted or displayed, butyl esters, and butanol. 
     
     
         48 . An animal feed product comprising the recombinant host cell of  claim 24 . 
     
     
         49 . The recombinant host cell of  claim 30 , wherein the alcohol production pathway is a butanol production pathway. 
     
     
         50 . The recombinant host cell of  claim 49  wherein the butanol production pathway is an isobutanol production pathway. 
     
     
         51 . The recombinant host cell of  claim 30  wherein the host cell further comprises reduced or eliminated pyruvate decarboxylase activity. 
     
     
         52 . An animal feed product comprising the recombinant host cell of  claim 30 .