In situ expression of lipase for enzymatic production of alcohol esters during fermentation
AssigneeButamax(Tm) Advanced Biofuels LLC
InventorsRobert DICOSIMO, Arthur Leo Kruckeberg, Thomas Edwin Van Aken
Life of this patent
Estimating the term…
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.
Claims48
A method comprising:
- a) providing a fermentation medium comprising fermentable carbon substrate derived from a biomass feedstock, alcohol produced from a fermentable carbon substrate derived from a biomass feedstock, and an alcohol-producing yeast microorganism wherein the alcohol-producing microorganism comprises an engineered polynucleotide encoding a polypeptide having lipase activity and the microorganism expresses and displays or secretes said polypeptide such that the lipase activity is present in the fermentation medium;
- b) contacting the fermentation medium with a carboxylic acid
- wherein the lipase activity is present in the fermentation medium in sufficient amount to convert at least a portion of the alcohol produced by the microorganism to alcohol esters extracellularly
(canceled)
(canceled)
(canceled)
A recombinant host cell comprising:
- (a) an engineered alcohol production pathway;
- and
- (b) an engineered polynucleotide encoding a polypeptide having lipase activity.
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.
A method of increasing tolerance of an alcohol-producing microorganism to the produced alcohol, the method comprising:
- (a) engineering a microorganism to express and secrete or display a polypeptide having lipase activity;
- (b) contacting the engineered microorganism with
- i. triglycerides, diglycerides, monoglycerides, phospholipids, free fatty acids, or a mixture thereof;
- and
- ii. a carbon substrate;
- under conditions whereby the microorganism produces an alcohol.
(canceled)
A method of producing butyl esters during a fermentation comprising
- (a) providing a fermentation medium comprising a carbon substrate and triglycerides, diglycerides, monoglycerides, and phospholipids, or a mixture thereof;
- and
- (b) contacting the fermentation medium with an alcohol-producing microorganism comprising a butanol biosynthetic pathway wherein said microorganism further comprises an engineered polynucleotide encoding a polypeptide having lipase activity and which expresses and secretes or displays the polypeptide such that the lipase activity is present in the fermentation medium.
(canceled)
(canceled)
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.
Dependencies are inferred from the claim text.