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<p>This research presents a method for the positive selection of mutants with improved xylose and L-arabinose fermentation in the thermotolerant, naturally xylose-utilizing yeast <i>Ogataea polymorpha</i> which is based on isolation of the mutants growing on L-arabinose as sole carbon and energy source. Whole-genome sequencing of the most efficient xylose-fermenting strain, A107, revealed mutations in the <i>API1</i> and <i>IRA1</i> genes, which are homologous to bacterial arabinose-5-phosphate isomerase and the Ras-GTPase activating domain in <i>Saccharomyces cerevisiae</i>, respectively. Disruption of the <i>IRA1</i> gene increased ethanol production during the fermentation of xylose and L-arabinose in <i>O. polymorpha</i> at 45&#xa0;°C. Overexpression of the <i>API1</i> gene specifically enhanced L-arabinose fermentation without affecting xylose fermentation. The most productive mutant strain accumulated 20.91&#xa0;g/L of ethanol in a xylose-containing medium at 45&#xa0;°C, exceeding the ethanol accumulation level of the wild-type strain (0.40&#xa0;g/L) by over 50 times. This strain holds potential for application in simultaneous saccharification and fermentation (SSF) processes.</p>

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Positive selection of efficient ethanol producers from xylose at 45 °C in the yeast Ogataea polymorpha

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年7月22日

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  • Roksolana Vasylyshyn,
  • Justyna Ruchala,
  • Kostyantyn Dmytruk,
  • Andriy Sibirny

摘要

This research presents a method for the positive selection of mutants with improved xylose and L-arabinose fermentation in the thermotolerant, naturally xylose-utilizing yeast Ogataea polymorpha which is based on isolation of the mutants growing on L-arabinose as sole carbon and energy source. Whole-genome sequencing of the most efficient xylose-fermenting strain, A107, revealed mutations in the API1 and IRA1 genes, which are homologous to bacterial arabinose-5-phosphate isomerase and the Ras-GTPase activating domain in Saccharomyces cerevisiae, respectively. Disruption of the IRA1 gene increased ethanol production during the fermentation of xylose and L-arabinose in O. polymorpha at 45 °C. Overexpression of the API1 gene specifically enhanced L-arabinose fermentation without affecting xylose fermentation. The most productive mutant strain accumulated 20.91 g/L of ethanol in a xylose-containing medium at 45 °C, exceeding the ethanol accumulation level of the wild-type strain (0.40 g/L) by over 50 times. This strain holds potential for application in simultaneous saccharification and fermentation (SSF) processes.