Molecular cloning, characterization, and engineering of xylitol dehydrogenase from Debaryomyces hansenii
- 11 April 2012
- journal article
- Published by Springer Science and Business Media LLC in Applied Microbiology and Biotechnology
- Vol. 97 (4), 1613-1623
- https://doi.org/10.1007/s00253-012-4020-5
Abstract
Because of its natural ability to utilize both xylose and arabinose, the halotolerant and osmotolerant yeast Debaryomyces hansenii is considered as a potential microbial platform for exploiting lignocellulosic biomass. To gain better understanding of the xylose metabolism in D. hansenii, we have cloned and characterized a xylitol dehydrogenase gene (DhXDH). The cloned gene appeared to be essential for xylose metabolism in D. hansenii as the deletion of this gene abolished the growth of the cells on xylose. The expression of DhXDH was strongly upregulated in the presence of xylose. Recombinant DhXdhp was expressed and purified from Escherichia coli. DhXdhp was highly active against xylitol and sorbitol as substrate. Our results showed that DhXdhp was thermo-sensitive, and except this, its biochemical properties were quite comparable with XDH from other yeast species. Furthermore, to make this enzyme suitable for metabolic engineering of D. hansenii, we have improved its thermotolerance and modified cofactor requirement through modelling and mutagenesis approach.Keywords
This publication has 43 references indexed in Scilit:
- Cloning and characterization of thermotolerant xylitol dehydrogenases from yeast Pichia angustaApplied Microbiology and Biotechnology, 2010
- Medium- and short-chain dehydrogenase/reductase gene and protein familiesCellular and Molecular Life Sciences, 2008
- Debaryomyces hansenii — an extremophilic yeast with biotechnological potentialYeast, 2006
- Role of N-Terminal Hydrophobic Region in Modulating the Subcellular Localization and Enzyme Activity of the Bisphosphate Nucleotidase from Debaryomyces hanseniiEukaryotic Cell, 2006
- Characterization of the AXDH gene and the encoded xylitol dehydrogenase from the dimorphic yeast Arxula adeninivoransAntonie van Leeuwenhoek, 2005
- Crystal structure of the NADP(H)-dependent ketose reductase from Bemisia argentifolii at 2.3 Å resolutionJournal of Molecular Biology, 2001
- Isolation and sequence of theHOG1 homologue fromDebaryomyces hansenii by complementation of thehog1? strain ofSaccharomyces cerevisiaeYeast, 2000
- Gapped BLAST and PSI-BLAST: a new generation of protein database search programsNucleic Acids Research, 1997
- Production of Xylitol from D-Xylose byDebaryomyces hanseniiApplied Biochemistry and Biotechnology, 1997
- Structural consequences of sequence patterns in the fingerprint region of the nucleotide binding fold: Implications for nucleotide specificityJournal of Molecular Biology, 1992