Importance

Loading Importance Indicators...

Abstract

A method for preparing a yeast capable of simultaneously fermenting a pentose and a hexose sugar, comprising: (a) providing a yeast comprising: one or more heterologous genes encoding an enzyme of a pentose metabolic pathway; disruptions of a gene encoding a ribulose-phosphate 3-epimerase and of a gene encoding a glucose-6-phosphate isomerase; one or more overexpressed endogenous genes encoding an enzyme of the pentose phosphate pathway; and a disruption of one or more genes encoding an NADPH dependent 6-phosphogluconate dehydrogenase, (b) subjecting said yeast to evolutionary engineering on a medium comprising a hexose sugar and at least one pentose sugar, selecting for a yeast with improved growth rate obtain an evolved yeast; (d) restoring, in the evolved yeast, one or more of the disrupted genes, or: (d′) identifying genetic permutations in at least part of the genome of the evolved yeast by genome sequencing; (e) constructing an improved pentose and hexose-fermenting yeast comprising one or more said genetic permutations. Also described is a recombinant yeast comprising one or more heterologous genes of a pentose metabolic pathway, and a gene encoding a variant of a parent polypeptide, the variant comprising an amino acid sequence comprising at least one mutation, when aligned with the amino acid sequence in SEQ ID NO: 6.

Loading the Abstract Image...

First Claim