title: Promising properties of a formate dehydrogenase from a methanol-assimilating yeast Ogataea parapolymorpha DL-1 in His-tagged form.
abstract: The cDNA gene coding for formate dehydrogenase (FDH) from Ogataea parapolymorpha DL-1 was cloned and expressed in Escherichia coli. The recombinant enzyme was purified by nickel affinity chromatography and was characterized as a homodimer composed of two identical subunits with approximately 40 kDa in each monomer. The enzyme showed wide pH optimum of catalytic activity from pH 6.0 to 7.0. It had relatively high optimum temperature at 65 °C and retained 93, 88, 83, and 71 % of its initial activity after 4 h of exposure at 40, 50, 55, and 60 °C, respectively, suggesting that this enzyme had promising thermal stability. In addition, the enzyme was characterized to have significant tolerance ability to organic solvents such as dimethyl sulfoxide, n-butanol, and n-hexane. The Michaelis-Menten constant (K(m)), turnover number (k(cat)), and catalytic efficiency (k(cat)/K(m)) values of the enzyme for the substrate sodium formate were estimated to be 0.82 mM, 2.32 s(-1), and 2.83 mM(-1) s(-1), respectively. The K m for NAD(+) was 83 μM. Due to its wide pH optimum, promising thermostability, and high organic solvent tolerance, O. parapolymorpha FDH may be a good NADH regeneration catalyst candidate.
authors.affiliation: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, 214122, China.
authors.forename: Shuhuai
authors.initials: S
authors.lastname: Yu
authors.forename: Lanjun
authors.initials: L
authors.lastname: Zhu
authors.forename: Chen
authors.initials: C
authors.lastname: Zhou
authors.forename: Tao
authors.initials: T
authors.lastname: An
authors.forename: Tao
authors.initials: T
authors.lastname: Zhang
authors.forename: Bo
authors.initials: B
authors.lastname: Jiang
authors.forename: Wanmeng
authors.initials: W
authors.lastname: Mu
mesh_terms: D002384:Catalysis; D005560:Formate Dehydrogenases / Q000378:metabolism*; D000432:Methanol / Q000378:metabolism*; D004718:Saccharomycetales / Q000201:enzymology*
chemical_list: D005560:Formate Dehydrogenases; D000432:Methanol