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| Cloning,Expression and Conversion Analysis of the Xylitol Dehydrogenase from Gluconobacter oxydans |
| SHEN Xiao-bo1,QI Xiang-hui1,2,ZHU Hong-yang1,XU Hong1 |
1.State Key Laboratory of MaterialsOriented Chemical Engineering,College of Food Science and Light Industry，Nanjing University of Technology，Nanjing 210009，China
2.School of Food and Biological Engineering，Jiangsu University，Zhenjiang 212013，China |
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Abstract Xdh encoding xylitol dehydrogenase(XDH)gene was amplified from the genome DNA of G.oxydans NH-10.The recombinant plasmid pSE-xdh was constructed by inserting xdh genes into expression vector pSE380 and transformed into E.coli JM109.The recombined XDH was purified through two steps including HisTrap HP affinity chromatography and SephacrylS-300 gel filtration chromatography,and then the enzymatic properties were investigated.The optimum pH and temperature for reduction conditions of XDH were 5.0 and 35℃,while that for the oxidation conditions were pH 11.0 and 30℃.XDH was a kind of NADH-dependent dehydrogenase,and the Km value for NADH was 57.8mmol/L and the Vmax was 1209.1mmol/(ml·min).The XDH activity of recombinant strain was 13.9 U/mg.16.7 g/L xylitol was obtained from 28.0 g/L D-xylulose in 16 h by mixed fermentation of resting cells which was composed of original strain and recombinant strain,whereas control strain produced 8.3 g/L xylitol.These results demonstrated that increasing XDH activity could improve xylitol productivity.
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Received: 09 September 2009
Published: 21 December 2009
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