Applied Microbiology and Biotechnology

, Volume 88, Issue 2, pp 497–507

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

  • Surisa Suwannarangsee
  • Doo-Byoung Oh
  • Jeong-Woo Seo
  • Chul Ho Kim
  • Sang Ki Rhee
  • Hyun Ah Kang
  • Warawut Chulalaksananukul
  • Ohsuk Kwon
Biotechnologically Relevant Enzymes and Proteins

DOI: 10.1007/s00253-010-2752-7

Cite this article as:
Suwannarangsee, S., Oh, DB., Seo, JW. et al. Appl Microbiol Biotechnol (2010) 88: 497. doi:10.1007/s00253-010-2752-7

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.

Keywords

Alcohol dehydrogenase·ADH1 Hansenula polymorpha Ethanol production Glycerol fermentation 

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Surisa Suwannarangsee
    • 1
  • Doo-Byoung Oh
    • 1
  • Jeong-Woo Seo
    • 2
  • Chul Ho Kim
    • 2
  • Sang Ki Rhee
    • 3
  • Hyun Ah Kang
    • 4
  • Warawut Chulalaksananukul
    • 5
  • Ohsuk Kwon
    • 1
  1. 1.Integrative Omics Research CenterKorea Research Institute of Bioscience and BiotechnologyDaejeonRepublic of Korea
  2. 2.Microbe-based Fusion Technology Research CenterJeonbuk Branch Institute, KRIBBJeonbukRepublic of Korea
  3. 3.Department of Medical BiotechnologySoonchunhyang UniversityChungnamRepublic of Korea
  4. 4.Department of Life Science, College of Natural ScienceChung-Ang UniversitySeoulRepublic of Korea
  5. 5.Biofuel Production by Biocatalyst Research Unit and Department of Botany, Faculty of ScienceChulalongkorn UniversityBangkokThailand