Applied Microbiology and Biotechnology

, 82:1067

Xylose isomerase from polycentric fungus Orpinomyces: gene sequencing, cloning, and expression in Saccharomyces cerevisiae for bioconversion of xylose to ethanol

  • Anjali Madhavan
  • Sriappareddy Tamalampudi
  • Kazunari Ushida
  • Daisuke Kanai
  • Satoshi Katahira
  • Aradhana Srivastava
  • Hideki Fukuda
  • Virendra S. Bisaria
  • Akihiko Kondo
Biotechnologically Relevant Enzymes and Proteins

DOI: 10.1007/s00253-008-1794-6

Cite this article as:
Madhavan, A., Tamalampudi, S., Ushida, K. et al. Appl Microbiol Biotechnol (2009) 82: 1067. doi:10.1007/s00253-008-1794-6

Abstract

The cDNA sequence of the gene for xylose isomerase from the rumen fungus Orpinomyces was elucidated by rapid amplification of cDNA ends. The 1,314-nucleotide gene was cloned and expressed constitutively in Saccharomyces cerevisiae. The deduced polypeptide sequence encoded a protein of 437 amino acids which showed the highest similarity to the family II xylose isomerases. Further, characterization revealed that the recombinant enzyme was a homodimer with a subunit of molecular mass 49 kDa. Cell extract of the recombinant strain exhibited high specific xylose isomerase activity. The pH optimum of the enzyme was 7.5, while the low temperature optimum at 37°C was the property that differed significantly from the majority of the reported thermophilic xylose isomerases. In addition to the xylose isomerase gene, the overexpression of the S. cerevisiae endogenous xylulokinase gene and the Pichia stipitis SUT1 gene for sugar transporter in the recombinant yeast facilitated the efficient production of ethanol from xylose.

Keywords

Xylose isomeraseOrpinomycesXylulokinaseSUT1Recombinant Saccharomyces cerevisiaeXylose fermentationEthanol

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Anjali Madhavan
    • 1
  • Sriappareddy Tamalampudi
    • 2
  • Kazunari Ushida
    • 3
  • Daisuke Kanai
    • 4
  • Satoshi Katahira
    • 2
  • Aradhana Srivastava
    • 1
  • Hideki Fukuda
    • 2
  • Virendra S. Bisaria
    • 1
  • Akihiko Kondo
    • 5
  1. 1.Department of Biochemical Engineering and BiotechnologyIndian Institute of Technology DelhiDelhiIndia
  2. 2.Organization of Advanced Science and TechnologyKobe UniversityKobeJapan
  3. 3.Laboratory of Animal ScienceKyoto Prefectural UniversityKyotoJapan
  4. 4.Research and Development Laboratories for Sustainable Value CreationAsahi Breweries, LtdIbarakiJapan
  5. 5.Department of Chemical Science and Engineering, Graduate School of EngineeringKobe UniversityKobeJapan