Engineered microbes and methods for microbial oil production

摘要

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.

主权项

1. 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 first nucleic acid construct comprising an expression cassette comprising a coding nucleic acid encoding the DGA1 gene product; and a second nucleic acid construct comprising an expression cassette comprising a coding nucleic acid encoding the ACC 1 gene product; wherein the first nucleic acid construct and the second nucleic acid construct further comprises an intron-enhanced promoter that comprises a promoter sequence, a transcription start site, and an intronic sequence downstream of the transcription start site, and wherein the promoter is a translation elongation factor-1α (TEF) promoter.