Characterization of alcohol dehydrogenase 1 of the thermotolerant methylotrophic yeast Hansenula polymorpha |
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Authors: | Surisa Suwannarangsee Doo-Byoung Oh Jeong-Woo Seo Chul Ho Kim Sang Ki Rhee Hyun Ah Kang Warawut Chulalaksananukul Ohsuk Kwon |
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Institution: | (1) Integrative Omics Research Center, Korea Research Institute of Bioscience and Biotechnology, 52 Eoeun-dong, Yuseong-gu, Daejeon, 305-333, Republic of Korea;(2) Microbe-based Fusion Technology Research Center, Jeonbuk Branch Institute, KRIBB, Jeonbuk, 580-185, Republic of Korea;(3) Department of Medical Biotechnology, Soonchunhyang University, Asan, Chungnam, 336-745, Republic of Korea;(4) Department of Life Science, College of Natural Science, Chung-Ang University, Seoul, 156-756, Republic of Korea;(5) Biofuel Production by Biocatalyst Research Unit and Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand; |
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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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