Molecular cloning, characterization, and engineering of xylitol dehydrogenase from Debaryomyces hansenii.

Biswas, Dipanwita and Datt, Manish and Aggarwal, Monika and Mondal, Alok K (2012) Molecular cloning, characterization, and engineering of xylitol dehydrogenase from Debaryomyces hansenii. Applied microbiology and biotechnology. ISSN 1432-0614

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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.

Item Type:	Article
Additional Information:	Copyright of this article belongs to Springer.
Subjects:	Q Science > QR Microbiology
Depositing User:	Dr. K.P.S.Sengar
Date Deposited:	28 Jan 2013 09:49
Last Modified:	28 Jan 2013 09:49
URI:	http://crdd.osdd.net/open/id/eprint/1251

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