Engineering of an endogenous hexose transporter into a specific D-xylose transporter facilitates glucose-xylose co-consumption in Saccharomyces cerevisiae

Jeroen G. Nijland, Hyun Yong Shin, Rene M. de Jong, Paul P. De Waal, Paul Klaassen, Arnold J. M. Driessen*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

84 Citations (Scopus)
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Background: Engineering of Saccharomyces cerevisiae for the simultaneous utilization of hexose and pentose sugars is vital for cost-efficient cellulosic bioethanol production. This yeast lacks specific pentose transporters and depends on endogenous hexose transporters for low affinity pentose uptake. Consequently, engineered xylose-fermenting yeast strains first utilize D-glucose before D-xylose can be transported and metabolized.

Results: We have used an evolutionary engineering approach that depends on a quadruple hexokinase deletion xylose-fermenting S. cerevisiae strain to select for growth on D-xylose in the presence of high D-glucose concentrations. This resulted in D-glucose-tolerant growth of the yeast of D-xylose. This could be attributed to mutations at N367 in the endogenous chimeric Hxt36 transporter, causing a defect in D-glucose transport while still allowing specific uptake of D-xylose. The Hxt36-N367A variant transports D-xylose with a high rate and improved affinity, enabling the efficient co-consumption of D-glucose and D-xylose.

Conclusions: Engineering of yeast endogenous hexose transporters provides an effective strategy to construct glucose-insensitive xylose transporters that are well integrated in the carbon metabolism regulatory network, and that can be used for efficient lignocellulosic bioethanol production.

Original languageEnglish
Article number168
Number of pages11
JournalBiotechnology for Biofuels
Issue number168
Publication statusPublished - 29-Nov-2014


  • Sugar transporter
  • Xylose transporter
  • Evolutionary engineering
  • Bioethanol
  • Yeast
  • KEY

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