title: Cloning and characterization of two distinct water-forming NADH oxidases from Lactobacillus pentosus for the regeneration of NAD.
abstract: Two uncharacterized nicotinamide adenine dinucleotide (NADH) oxidases (named as LpNox1, LpNox2) from Lactobacillus pentosus ATCC 8041 were cloned and overexpressed in Escherichia coli BL21 (DE3). The sequence analysis revealed that the two enzymes are water-forming Noxs with 64 % and 52 % identity to LbNox from Lactobacillus brevis DSM 20054. The optimal pH and temperature of the purified LpNox1 and LpNox2 were 7.0 and 8.0 and 35 and 40 °C, respectively, with K M of 99.0 μM (LpNox1) and 27.6 μM (LpNox2), and yielding catalytic efficiency k cat/K M of 1.0 and 0.2 μM(-1) s(-1), respectively. Heat inactivation studies revealed that the two enzymes are relatively instable. The application of LpNox1 for the regeneration of NAD(+) was demonstrated by coupling with a glycerol dehydrogenase-catalyzed oxidation of glycerol to 1,3-dihydroxyacetone. The characteristics of the LpNox1 could prove to be of interest in industrial application such as NAD(+) regeneration in dehydrogenase-catalyzed oxidations.
authors.affiliation: Department of Biological and Pharmaceutical Engineering, College of Chemistry and Chemical Engineering, Taiyuan University of Technology, No.79 West Yingze Street, Taiyuan, Shanxi, 030024, People's Republic of China. zhangjiandong@tyut.edu.cn.
authors.forename: Jian-Dong
authors.initials: JD
authors.lastname: Zhang
authors.affiliation: Department of Biological and Pharmaceutical Engineering, College of Chemistry and Chemical Engineering, Taiyuan University of Technology, No.79 West Yingze Street, Taiyuan, Shanxi, 030024, People's Republic of China.
authors.forename: Zhi-Mei
authors.initials: ZM
authors.lastname: Cui
authors.affiliation: Department of Biological and Pharmaceutical Engineering, College of Chemistry and Chemical Engineering, Taiyuan University of Technology, No.79 West Yingze Street, Taiyuan, Shanxi, 030024, People's Republic of China. fxjbio@163.com.
authors.forename: Xiao-Jun
authors.initials: XJ
authors.lastname: Fan
authors.affiliation: Department of Biological and Pharmaceutical Engineering, College of Chemistry and Chemical Engineering, Taiyuan University of Technology, No.79 West Yingze Street, Taiyuan, Shanxi, 030024, People's Republic of China.
authors.forename: Hua-Lei
authors.initials: HL
authors.lastname: Wu
authors.affiliation: Department of Biological and Pharmaceutical Engineering, College of Chemistry and Chemical Engineering, Taiyuan University of Technology, No.79 West Yingze Street, Taiyuan, Shanxi, 030024, People's Republic of China.
authors.forename: Hong-Hong
authors.initials: HH
authors.lastname: Chang
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D002384:Catalysis; D003001:Cloning, Molecular; D006863:Hydrogen-Ion Concentration; D000070018:Lactiplantibacillus pentosus / Q000201:enzymology / Q000235:genetics; D009243:NAD / Q000235:genetics / Q000378:metabolism*; D019255:NADPH Oxidases / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D010084:Oxidation-Reduction
chemical_list: D001426:Bacterial Proteins; D009243:NAD; D019255:NADPH Oxidases
keywords: Cloning; Cofactor regeneration; Lactobacillus pentosus; NADH oxidase; Water forming