title: A 2,3-butanediol dehydrogenase from Paenibacillus polymyxa ZJ-9 for mainly producing R,R-2,3-butanediol: purification, characterization and cloning.
abstract: A 2,3-butanediol dehydrogenase (BDH) from Paenibacillus polymyxa ZJ-9 was purified to homogeneity via fractional ammonium sulfate precipitation, followed by two steps of anion-exchange chromatography using DEAE-Sepharose and Source 15Q, obtaining a 35-fold increase in specific activity and 34.9% yield. The molecular weights of the purified BDH subunit and holoenzyme were 44.5 and 90.0 kDa, respectively, as detected via SDS-PAGE and gel filtration chromatography. These results were significantly different from those of other reported BDHs. Substrate specificity experiments showed that the enzyme could function preferentially as a reductase rather than as a dehydrogenase, and was mainly responsible for the reduction of R-acetoin to R,R-2,3-butanediol. Gene cloning, sequencing, and expression experiments further demonstrate that this enzyme was a new type of BDH.
authors.affiliation: State Key Laboratory of Materials-Oriented Chemical Engineering, College of Food Science and Light Industry, Nanjing University of Technology, Nanjing, P.R. China; School of Chemical and Biological Engineering, Yancheng Institute of Technology, Yancheng, P.R. China.
authors.forename: Jian
authors.initials: J
authors.lastname: Gao
authors.forename: Huan-Huan
authors.initials: HH
authors.lastname: Yang
authors.forename: Xiao-Hai
authors.initials: XH
authors.lastname: Feng
authors.forename: Sha
authors.initials: S
authors.lastname: Li
authors.forename: Hong
authors.initials: H
authors.lastname: Xu
mesh_terms: D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D002072:Butylene Glycols / Q000378:metabolism*; D005591:Chemical Fractionation; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D008970:Molecular Weight; D056507:Paenibacillus / Q000201:enzymology* / Q000235:genetics; D055503:Protein Multimerization; D013379:Substrate Specificity
chemical_list: D002072:Butylene Glycols; C026978:2,3-butylene glycol; D000429:Alcohol Oxidoreductases; C019801:butanediol dehydrogenase