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| Short-chain D-arabitol Dehydrogenase from Gluconobacter oxydans NH-10 |
| DAI Xiao-yan, SHEN Xiao-bo, ZHU Hong-yang, XU Hong |
| State Key Laboratory of Materials-Oriented Chemical Engineering, College of Food Science and Light Industry, Nanjing University of Technology,Nanjing 210009, China |
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Abstract Based on combination of bioinformatics, The similarity between the cloning gene and corresponding gene of Acetobacter suboxydans reach to 80%. Then, recombinant plasmid was constructed by inserting arDH genes into vector pET22b and functionally expressed into E. coli JM109(DE3). The molecular mass of recombinant D-arabitol dehydrogenase was about 30 kDa, so it belonged to the short-chain dehydrogenase. The recombinant enzyme was purified by His Trap and subjected to enzymological characterization. The ArDH could not only oxidize D-arabitol to D-xylulose, but also reduce D-xylulose to D-arabitol. When catalyzing oxidative reaction, the Km value for D-arabitol was 60.67 mmol/L, Vmax was 0.803 U/mg; it could use NAD+ and NADP+ as cozyme, but it preferred to depending on NAD+; the optimum pH and temperature of oxidative reactions was 12.0 and 30℃. However, when catalyzing reductive reactions, the Km value for D-xylulose was 36.93 mmol/L, Vmax was 1.71 U/mg; the optimum pH and temperature of reductive reactions was 7.0 and 30℃.
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Published: 19 November 2010
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