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United States Department of Agriculture

Agricultural Research Service

Title: Expression of a heterologous xylose transporter in a Saccharomyces cerevisiae strain engineered to utilize xylose increases xylose uptake and improves xylose/glucose co-consumption

Authors
item Hector, Ronald
item Qureshi, Nasib
item Hughes, Stephen
item Cotta, Michael

Submitted to: Biotechnology for Fuels and Chemicals Symposium Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: May 7, 2008
Publication Date: May 17, 2008
Citation: Hector, R.E., Qureshi, N., Hughes, S.R., Cotta, M.A. 2008. Expression of a heterologous xylose transporter in a Saccharomyces cerevisiae strain engineered to utilize xylose increases xylose uptake and improves xylose/glucose co-consumption [abstract]. In: Proceedings of the 30th Symposium on Biotechnology for Fuels and Chemicals, May 4-7, 2008, New Orleans, LA. p. 131.

Technical Abstract: Strains of Saccharomyces cerevisiae have been engineered to utilize xylose by expressing either the genes for xylose reductase and xylitol dehydrogenase, or for xylose isomerase. These strains still use xylose at sub-optimal rates for industrial fermentation. Unlike natural xylose fermenting yeasts such as Pichia stipitis, S. cerevisiae does not contain xylose-specific transport systems. Instead, xylose enters S. cerevisiae cells through hexose transporters. Analysis of strains that have been adapted for enhanced growth on xylose indicates increased expression of hexose transporters and suggests that xylose transport is one of the limiting steps in xylose utilization. Since the increase in xylose transport in these strains results from increases in hexose transporters, xylose uptake remains inhibited by glucose, thus limiting xylose and glucose co-consumption. The goal of this study was to determine the effect of a xylose transport system on xylose and glucose co-consumption as well as total xylose consumption. We expressed two heterologous transporters from Arabidopsis thaliana in S. cerevisiae strains that were previously engineered to utilize xylose. Strains expressing the heterologous transporters were grown aerobically on glucose and xylose mixtures. Sugar consumption rates and product accumulation were determined and compared to a control strain not expressing the A. thaliana xylose transport genes. Our results from aerobic cultivation with glucose/xylose mixtures indicate that expression of the transporters increased xylose co-consumption rates (prior to glucose depletion) by up to 100%. Increased xylose co-consumption also correlated with increased ethanol concentration, yield, and specific productivity. It was concluded that in these strains, xylose transport is a limiting factor for xylose utilization and that increasing xylose/glucose co-consumption is a viable strategy for improving xylose fermentation.

Last Modified: 10/21/2014
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