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Issue 11, 2010
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Discovery and characterization of novel D-xylose-specific transporters from Neurospora crassa and Pichia stipitis

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Abstract

Saccharomyces cerevisiae is considered one of the most promising organisms for ethanol production from lignocellulosic feedstock. Unfortunately, pentose sugars, which comprise up to 30% of lignocellulose, cannot be utilized by wild type S. cerevisiae. Heterologous pathways were introduced into S. cerevisiae to enable utilization of D-xylose, the most abundant pentose sugar. However, the resulting recombinant S. cerevisiae strains exhibited a slow growth rate and poor sugar utilization efficiency when grown on D-xylose as the sole carbon source. D-xylose uptake is the first step of D-xylose utilization. D-xylose can only enter yeast cells through hexose transporters, which have two orders of magnitude lower affinity towards D-xylose compared to hexoses. It was also shown that inefficient pentose uptake is the limiting step in some D-xylose metabolizing yeast strains. Here we report the cloning and characterization of two novel D-xylose-specific transporters from Neurospora crassa and Pichia stipitis. These two transporters were identified from a total of 18 putative pentose transporters. They were functionally expressed and properly localized in S. cerevisiae as indicated by HPLC analysis and fluorescence confocal microscopy, respectively. Kinetic parameters of the D-xylose-specific transporters were determined using a 14C-labeled sugar uptake assay. Use of pentose-specific transporters should improve D-xylose consumption and ethanol production in fast D-xylose assimilating strains, thereby lowering the cost of lignocellulosic ethanol production.

Graphical abstract: Discovery and characterization of novel d-xylose-specific transporters from Neurospora crassa and Pichia stipitis

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Publication details

The article was received on 11 May 2010, accepted on 09 Jun 2010 and first published on 11 Aug 2010


Article type: Paper
DOI: 10.1039/C0MB00007H
Citation: Mol. BioSyst., 2010,6, 2150-2156
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    Discovery and characterization of novel D-xylose-specific transporters from Neurospora crassa and Pichia stipitis

    J. Du, S. Li and H. Zhao, Mol. BioSyst., 2010, 6, 2150
    DOI: 10.1039/C0MB00007H

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