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Xylose Fermentation Was Improved by Kluyveromyces marxianus KHM89 through Up-regulation of Nicotinamide Adenine Dinucleotide (NAD+) Salvage Pathway

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Abstract

Xylose fermentation has been reported to be improved in Kluyveromyces marxianus via strain improvement by overexpressing xylose reductase (XYL1) and xylitol dehydrogenase (XYL2). This study performed directed evolution to further enhance xylose consumption in a K. marxianus mutant following transcriptomic analysis to determine genes associated with enhanced characteristics. KmXYL1 and KmXYL2 genes were overexpressed in K. marxianus 17555ΔURA3 for improving xylose fermentation. By performing directed evolution, a mutant K. marxianus KHM89 showing enhanced ethanol production was isolated from xylose medium. K. marxianus KHM89 consumed 47.39 g/L of xylose and produced 22.62 g/L of xylitol and 10.59 g/L of ethanol while the parental strain consumed 25.15 g/L of xylose and produced 7.36 g/L of xylitol and 2.05 g/L of ethanol. RNA sequencing-based transcriptomic analysis showed that alcohol dehydrogenases, aldehyde dehydrogenases, and NAD+ salvage pathway enzymes were upregulated in K. marxianus KHM89. These results were achieved via a combinatorial approach of rational design and directed evolution. The findings of this study contribute to the improvement of xylose fermentation by K. marxianus at an industrial scale.

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Acknowledgements

This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2019R1F1A1055315).

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Authors and Affiliations

  1. Department of Bioengineering and Technology, Kangwon National University, Chuncheon, 24341, Korea

    Deok-Ho Kwon & Suk-Jin Ha

  2. Department of Biohealth-machinery Convergence Engineering, Kangwon National University, Chuncheon, 24341, Korea

    Deok-Ho Kwon & Suk-Jin Ha

  3. Institute of Fermentation and Brewing, Kangwon National University, Chuncheon, 24341, Korea

    Suk-Jin Ha

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  1. Deok-Ho Kwon
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Deok-Ho Kwon contributed to the study conception, design, material preparation and data collection. Suk-Jin Ha contributed to review and editing of the manuscript.

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Correspondence to Suk-Jin Ha.

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Kwon, DH., Ha, SJ. Xylose Fermentation Was Improved by Kluyveromyces marxianus KHM89 through Up-regulation of Nicotinamide Adenine Dinucleotide (NAD+) Salvage Pathway. Biotechnol Bioproc E 27, 624–630 (2022). https://doi.org/10.1007/s12257-022-0036-6

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  • DOI: https://doi.org/10.1007/s12257-022-0036-6

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