Deletion of the GRE3 Aldose Reductase Gene and Its Influence on Xylose Metabolism in Recombinant Strains of Saccharomyces cerevisiae Expressing thexylA and XKS1 Genes

ABSTRACT

Saccharomyces cerevisiae ferments hexoses efficiently but is unable to ferment xylose. When the bacterial enzyme xylose isomerase (XI) from Thermus thermophilus was produced in S. cerevisiae, xylose utilization and ethanol formation were demonstrated. In addition, xylitol and acetate were formed. An unspecific aldose reductase (AR) capable of reducing xylose to xylitol has been identified inS. cerevisiae. The GRE3gene, encoding the AR enzyme, was deleted in S.cerevisiae CEN.PK2-1C, yielding YUSM1009a. XI fromT. thermophilus was produced, and endogenous xylulokinase from S.cerevisiae was overproduced in S.cerevisiae CEN.PK2-1C and YUSM1009a. In recombinant strains from which the GRE3 gene was deleted, xylitol formation decreased twofold. Deletion of the GRE3 gene combined with expression of the xylA gene fromT. thermophilus on a replicative plasmid generated recombinant xylose utilizing S.cerevisiae strain TMB3102, which produced ethanol from xylose with a yield of 0.28 mmol of C from ethanol/mmol of C from xylose. None of the recombinant strains grew on xylose.