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Cloning and Overexpression of the Xylitol Dehydrogenase Gene from Bacillus pallidusand Its Application to L-Xylulose Production
- Source :
- Bioscience, Biotechnology, and Biochemistry; September 2010, Vol. 74 Issue: 9 p1807-1813, 7p
- Publication Year :
- 2010
-
Abstract
- The xylitol dehydrogenase gene (xdh) of Bacillus palliduswas cloned and overexpressed in Escherichia coliusing pQE60 vector, for the first time. The open reading frame of 759 bp encoded a 253 amino acid protein with a calculated molecular mass of 27,333 Da. The recombinant xylitol dehydrogenase (XDH) was purified to homogeneity by three-step column chromatography, producing a single SDS–PAGE band of 28 kDa apparent molecular mass. The enzyme exhibited maximal activity at 55 °C in glycine-NaOH buffer pH 11.0, with 66% of initial enzyme activity retained after incubation at 40 °C for 1 h. In further application of the recombinant bacterium to L-xylulose production from xylitol (initial concentration 5%) using a resting cell reaction, 35% L-xylulose was produced within 24 h. This result indicates that this recombinant XDH is applicable in the large-scale production of L-xylulose.
Details
- Language :
- English
- ISSN :
- 09168451 and 13476947
- Volume :
- 74
- Issue :
- 9
- Database :
- Supplemental Index
- Journal :
- Bioscience, Biotechnology, and Biochemistry
- Publication Type :
- Periodical
- Accession number :
- ejs32896937
- Full Text :
- https://doi.org/10.1271/bbb.100144