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Construction of Industrial Saccharomyces cerevisiae Expressing Xylose-Metabolizing Genes in XI Pathway

In order to develop an industrial strain of Saccharomyces cerevisiae that can utilize the xylose to produce ethanol, an integrating plasmid pYMIK-xy114 containing the xylA gene from Thermus thermophilus, encoding xylose isomerase (XI), and XKS1 gene from S. cerevisiae, encoding xylulokinase (XK) was constructed. The integrating plasmid pYMIK-xy114 was transformed into a S. cerevisiae industrial strain NAN-27 producing the recombinant strain NAN-114. Upon transformation, multiple copies of the xylose-utilizing genes were integrated into the genome rDNA locus of S. cerevisiae. The results of enzymes assays showed that the XI and XK activity of NAN-114 was higher than that of parent strain. The resulting recombinant strain could co-ferment glucose and xylose to ethanol under oxygen-limited condition. The NAN-114 consumed 4.6g/L xylose and produced 6.9g/L ethanol, which were 43.8% and 9.5% higher than the parent strain NAN-27 respectively.