Engineered microbes and methods for microbial oil production
AssigneeMassachusetts Institute of Technology
InventorsGregory Stephanopoulos, Mitchell Tai, Sagar Chakraborty
Life of this patent
Estimating the term…
Abstract
Some aspects of this invention provide engineered microbes for oil production. Methods for microbe engineering and for use of engineered microbes are also provided herein. In some embodiments, microbes are provided that are engineered to modulate a combination of rate-controlling steps of lipid synthesis, for example, a combination of a step generating metabolites, acetyl-CoA, ATP or NADPH for lipid synthesis (a push step), and a step sequestering a product or an intermediate of a lipid synthesis pathway that mediates feedback inhibition of lipid synthesis (a pull step). Such push-and-pull engineered microbes exhibit greatly enhanced conversion yields and TAG synthesis and storage properties.
Claims15
An isolated oleaginous yeast cell, comprising a genetic modification that increases expression of a diacylglycerol acyltransferase (DGA1) gene product and an acetyl-CoA carboxylase (ACC1) gene product, wherein the genetic modification comprises
- (a) a first nucleic acid construct comprising an expression cassette comprising a coding nucleic acid encoding the DGA1 gene product,
- wherein the coding nucleic acid encoding the DGA1 gene product comprises a nucleic acid sequence that is at least 85% identical to SEQ ID NO:
- 1,
- wherein the first nucleic acid construct comprises an intron-enhanced promoter;
- and
- (b) a second nucleic acid construct comprising an expression cassette comprising a coding nucleic acid encoding the ACC1 gene product,
- wherein the coding nucleic acid encoding the ACC1 gene product encodes for a protein comprising the amino acid sequence of SEQ ID NO:
- 4,
- wherein the isolated oleaginous yeast cell converts a carbon source to a fatty acid or a triacylglycerol (TAG) at a conversion rate within the range of about 0.025 g/g to about 0.32 g/g (g TAG produced/g Glucose consumed).
Dependencies are inferred from the claim text.