Kawano Shigeru, Yano Miho, Hasegawa Junzo, Yasohara Yoshihiko
Frontier Biochemical and Medicinal Research Laboratories, Kaneka Corporation, Takasago, Hyogo, Japan.
Biosci Biotechnol Biochem. 2011;75(6):1055-60. doi: 10.1271/bbb.100528. Epub 2011 Jun 13.
A novel (R)-specific alcohol dehydrogenase (AFPDH) produced by Candida maris IFO10003 was purified to homogeneity by ammonium sulfate fractionation, DEAE-Toyopearl, and Phenyl-Toyopearl, and characterized. The relative molecular mass of the native enzyme was found to be 59,900 by gel filtration, and that of the subunit was estimated to be 28,900 on SDS-polyacrylamide gel electrophoresis. These results suggest that the enzyme is a homodimer. It required NADH as a cofactor and reduced various kinds of carbonyl compounds, including ketones and aldehydes. AFPDH reduced acetylpyridine derivatives, β-keto esters, and some ketone compounds with high enantioselectivity. This is the first report of an NADH-dependent, highly enantioselective (R)-specific alcohol dehydrogenase isolated from a yeast. AFPDH is a very useful enzyme for the preparation of various kinds of chiral alcohols.
海生假丝酵母IFO10003产生的一种新型（R）特异性醇脱氢酶（AFPDH）通过硫酸铵分级分离、DEAE- Toyopearl和苯基-Toyopearl纯化至同质，并进行了表征。通过凝胶过滤发现天然酶的相对分子质量为59,900，在SDS-聚丙烯酰胺凝胶电泳上亚基的相对分子质量估计为28,900。这些结果表明该酶是同型二聚体。它需要NADH作为辅因子，并能还原各种羰基化合物，包括酮和醛。AFPDH以高对映选择性还原乙酰吡啶衍生物、β-酮酯和一些酮化合物。这是首次报道从酵母中分离出的依赖NADH的、高对映选择性的（R）特异性醇脱氢酶。AFPDH是制备各种手性醇的非常有用的酶。
