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Essentiality of respiratory activity for pentose utilization in thermotolerant yeast Kluyveromyces marxianus DMKU 3-1042

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Abstract

By random integrative mutagenesis with a kanMX4 cassette in Kluyveromyces marxianus DMKU 3-1042, we obtained three mutants of COX15, ATP25 and CYC3 encoding a cytochrome oxidase assembly factor (singleton), a transcription factor required for assembly of the Atp9p subunit of mitochondrial ATP synthase and cytochrome c heme lyase, respectively, as mutants lacking growth capability on xylose and/or arabinose. They exhibited incapability of growth on non-fermentable carbon sources, such as acetate or glycerol, and thermosensitiveness. Their biomass formation in glucose medium was reduced, but ethanol yields were increased with a high ethanol level in the medium, compared to those of the parental strain. Experiments with respiratory inhibitors showed that cox15 and cyc3, but not atp25, were able to grow in glucose medium containing antimycin A and that the atp25 mutant was KCN-resistant. Activities of NADH and ubiquinol oxidases in membrane fractions of each mutant became a half of that of the parent and negligible, respectively, and their remaining NADH oxidase activities were found to be resistant to KCN. Absolute absorption spectral analysis revealed that the peak corresponding to a + a 3 was very small in atp25 and negligible in cox15 and cyc3. These findings suggest that the K. marxianus strain possesses an alternative KCN-resistant oxidase that is located between primary dehydrogenases and the ubiquinone pool and that the respiratory activity is essential for utilization of pentoses.

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Acknowledgments

We thank Dr. Kazunobu Matsushita and Dr. Toshiharu Yakushi for their helpful discussion. This work was supported by the Program for Promotion of Basic Research Activities for Innovative Biosciences, New Energy and Industrial Technology Development Organization (NEDO) and the Special Coordination Funds for Promoting Science & Technology, Ministry of Education, Culture, Sports, Science & Technology. This work was performed as a collaborative research in the Asian Core Program between Yamaguchi University and Khon Kaen University, which was supported by the Scientific Cooperation Program agreed by the Japan Society for the Promotion of Science (JSPS) and the National Research Council of Thailand (NRCT).

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Authors and Affiliations

  1. Applied Molecular Bioscience, Graduate School of Medicine, Yamaguchi University, Ube, 755-8505, Japan

    Noppon Lertwattanasakul,  Suprayogi, Masayuki Murata & Mamoru Yamada

  2. Department of Microbiology, Faculty of Science, Kasetsart University, Bangkok, 10900, Thailand

    Noppon Lertwattanasakul & Savitree Limtong

  3. Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai, 50200, Thailand

    Nadchanok Rodrussamee

  4. Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi, 753-8515, Japan

    Tomoyuki Kosaka & Mamoru Yamada

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  1. Noppon Lertwattanasakul
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Correspondence to Mamoru Yamada.

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Lertwattanasakul, N., Suprayogi, Murata, M. et al. Essentiality of respiratory activity for pentose utilization in thermotolerant yeast Kluyveromyces marxianus DMKU 3-1042. Antonie van Leeuwenhoek 103, 933–945 (2013). https://doi.org/10.1007/s10482-012-9874-0

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