Saccharomyces cerevisiae Engineered for Xylose Metabolism Exhibits a Respiratory Response

Abstract

ABSTRACT Native strains of Saccharomyces cerevisiae do not assimilate xylose. S. cerevisiae engineered for d -xylose utilization through the heterologous expression of genes for aldose reductase ( XYL1 ), xylitol dehydrogenase ( XYL2 ), and d -xylulokinase ( XYL3 or XKS1 ) produce only limited amounts of ethanol in xylose medium. In recombinant S. cerevisiae expressing XYL1 , XYL2 , and XYL3 , mRNA transcript levels for glycolytic, fermentative, and pentose phosphate enzymes did not change significantly on glucose or xylose under aeration or oxygen limitation. However, expression of genes encoding the tricarboxylic acid cycle, respiration enzymes ( HXK1 , ADH2 , COX13 , NDI1 , and NDE1 ), and regulatory proteins ( HAP4 and MTH1 ) increased significantly when cells were cultivated on xylose, and the genes for respiration were even more elevated under oxygen limitation. These results suggest that recombinant S. cerevisiae does not recognize xylose as a fermentable carbon source and that respiratory proteins are induced in response to cytosolic redox imbalance; however, lower sugar uptake and growth rates on xylose might also induce transcripts for respiration. A petite respiration-deficient mutant (ρ°) of the engineered strain produced more ethanol and accumulated less xylitol from xylose. It formed characteristic colonies on glucose, but it did not grow on xylose. These results are consistent with the higher respiratory activity of recombinant S. cerevisiae when growing on xylose and with its inability to grow on xylose under anaerobic conditions.