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Characterization of alcohol dehydrogenase 1 of the thermotolerant methylotrophic yeast Hansenula polymorpha

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Abstract

The thermotolerant methylotrophic yeast Hansenula polymorpha has recently been gaining interest as a promising host for bioethanol production due to its ability to ferment xylose, glucose, and cellobiose at elevated temperatures up to 48 °C. In this study, we identified and characterized alcohol dehydrogenase 1 of H. polymorpha (HpADH1). HpADH1 seems to be a cytoplasmic protein since no N-terminal mitochondrial targeting extension was detected. Compared to the ADHs of other yeasts, recombinant HpADH1 overexpressed in Escherichia coli exhibited much higher catalytic efficiency for ethanol oxidation along with similar levels of acetaldehyde reduction. HpADH1 showed broad substrate specificity for alcohol oxidation but had an apparent preference for medium chain length alcohols. Both ADH isozyme pattern analysis and ADH activity assay indicated that ADH1 is the major ADH in H. polymorpha DL-1. Moreover, an HpADH1-deleted mutant strain produced less ethanol in glucose or glycerol media compared to wild-type. Interestingly, when the ADH1 mutant was complemented with an HpADH1 expression cassette, the resulting strain produced significantly increased amounts of ethanol compared to wild-type, up to 36.7 g l−1. Taken together, our results suggest that optimization of ADH1 expression would be an ideal method for developing H. polymorpha into an efficient bioethanol production strain.

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Acknowledgement

This work was supported by a grant from the KRIBB Research Initiative Program and by a National Research Foundation of Korea (NRF) grant funded by the Korean government (MEST) (No. 2009-0075186). S. Suwannarangsee was supported by the International Scholar Exchange Fellowship for the academic year of 2008–2009 by the Korea Foundation for Advanced Studies.

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Authors and Affiliations

  1. Integrative Omics Research Center, Korea Research Institute of Bioscience and Biotechnology, 52 Eoeun-dong, Yuseong-gu, Daejeon, 305-333, Republic of Korea

    Surisa Suwannarangsee, Doo-Byoung Oh & Ohsuk Kwon

  2. Microbe-based Fusion Technology Research Center, Jeonbuk Branch Institute, KRIBB, Jeonbuk, 580-185, Republic of Korea

    Jeong-Woo Seo & Chul Ho Kim

  3. Department of Medical Biotechnology, Soonchunhyang University, Asan, Chungnam, 336-745, Republic of Korea

    Sang Ki Rhee

  4. Department of Life Science, College of Natural Science, Chung-Ang University, Seoul, 156-756, Republic of Korea

    Hyun Ah Kang

  5. Biofuel Production by Biocatalyst Research Unit and Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand

    Warawut Chulalaksananukul

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  1. Surisa Suwannarangsee
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  2. Doo-Byoung Oh
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  3. Jeong-Woo Seo
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  4. Chul Ho Kim
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  5. Sang Ki Rhee
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  8. Ohsuk Kwon
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Correspondence to Ohsuk Kwon.

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Suwannarangsee, S., Oh, DB., Seo, JW. et al. Characterization of alcohol dehydrogenase 1 of the thermotolerant methylotrophic yeast Hansenula polymorpha . Appl Microbiol Biotechnol 88, 497–507 (2010). https://doi.org/10.1007/s00253-010-2752-7

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  • DOI: https://doi.org/10.1007/s00253-010-2752-7

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