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Molecular Cloning and Characterization of NAD~+-Dependent Xylitol Dehydrogenase from Candida tropicalis ATCC 20913

机译:热带假丝酵母ATCC 20913的NAD〜+依赖性木糖醇脱氢酶的分子克隆与表征

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摘要

Induction of xylitol dehydrogenase of Candida tropicalis ATCC 20913 by various carbon sources was investigated.The enzyme activity was induced when the yeast was grown on L-arabinose and D-xylose.A novel gene encoding the enzyme was cloned and characterized.The 1,095-bp coding sequence of the gene encodes a polypeptide of 364 amino acids,with a molecular mass of 39.4 kDa.Sequence analysis of the putative protein showed it to be a member of the zinc-containing alcohol dehydrogenase family and to have homology to xylitol dehydrogenase genes from other yeasts and fungi.The recombinant xylitol dehydrogenase expressed in Escherichia coli oxidized polyols such as xylitol and D-sorbitol and reduced ketoses such as D-xylulose and D-fructose.It required exclusively NAD or NADH as a cofactor.
机译:研究了不同碳源对热带念珠菌ATCC 20913的木糖醇脱氢酶的诱导作用,酵母在L-阿拉伯糖和D-木糖上生长后诱导了酶的活性,克隆并鉴定了一个新的编码该酶的基因,全长1,095bp该基因的编码序列编码一个364个氨基酸的多肽,分子量为39.4 kDa。推定蛋白质的序列分析表明,它是含锌醇脱氢酶家族的成员,并且与木糖醇脱氢酶基因具有同源性。在大肠杆菌中表达的重组木糖醇脱氢酶氧化了诸如木糖醇和D-山梨糖醇之类的多元醇以及诸如D-木酮糖和D-果糖之类的还原酮糖,仅需要NAD或NADH作为辅因子。

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