title: An efficient ribitol-specific dehydrogenase from Enterobacter aerogenes.
abstract: An NAD(+)-dependent ribitol dehydrogenase from Enterobacter aerogenes KCTC 2190 (EaRDH) was cloned and successfully expressed in Escherichia coli. The complete 729-bp gene was amplified, cloned, expressed, and subsequently purified in an active soluble form using nickel affinity chromatography. The enzyme had an optimal pH and temperature of 11.0 and 45°C, respectively. Among various polyols, EaRDH exhibited activity only toward ribitol, with Km, Vmax, and kcat/Km values of 10.3mM, 185Umg(-1), and 30.9s(-1)mM(-1), respectively. The enzyme showed strong preference for NAD(+) and displayed no detectable activity with NADP(+). Homology modeling and sequence analysis of EaRDH, along with its biochemical properties, confirmed that EaRDH belongs to the family of NAD(+)-dependent ribitol dehydrogenases, a member of short-chain dehydrogenase/reductase (SCOR) family. EaRDH showed the highest activity and unique substrate specificity among all known RDHs. Homology modeling and docking analysis shed light on the molecular basis of its unusually high activity and substrate specificity.
authors.affiliation: Department of Chemical Engineering, Konkuk University, 1 Hwayang-Dong, Gwangjin-Gu, Seoul 143-701, South Korea.
authors.forename: Ranjitha
authors.initials: R
authors.lastname: Singh
authors.affiliation: Department of Chemical Engineering, Konkuk University, 1 Hwayang-Dong, Gwangjin-Gu, Seoul 143-701, South Korea.
authors.forename: Raushan
authors.initials: R
authors.lastname: Singh
authors.affiliation: Department of Chemical Engineering, Konkuk University, 1 Hwayang-Dong, Gwangjin-Gu, Seoul 143-701, South Korea.
authors.forename: In-Won
authors.initials: IW
authors.lastname: Kim
authors.affiliation: Department of Chemical Engineering, Konkuk University, 1 Hwayang-Dong, Gwangjin-Gu, Seoul 143-701, South Korea.
authors.forename: Sujan
authors.initials: S
authors.lastname: Sigdel
authors.affiliation: Microbial Biotechnology and Genomics, CSIR-Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, Delhi 110007, India.
authors.forename: Vipin C
authors.initials: VC
authors.lastname: Kalia
authors.affiliation: Department of Materials Science and Engineering, Korea University, Anam-Dong, Seongbuk-Gu, Seoul 136-713, South Korea. Electronic address: yckang@korea.ac.kr.
authors.forename: Yun Chan
authors.initials: YC
authors.lastname: Kang
authors.affiliation: Department of Chemical Engineering, Konkuk University, 1 Hwayang-Dong, Gwangjin-Gu, Seoul 143-701, South Korea. Electronic address: jkrhee@konkuk.ac.kr.
authors.forename: Jung-Kul
authors.initials: JK
authors.lastname: Lee
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001709:Biotechnology; D020134:Catalytic Domain; D021902:Enterobacter aerogenes / Q000201:enzymology* / Q000235:genetics; D005798:Genes, Bacterial; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008970:Molecular Weight; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012255:Ribitol / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D040681:Structural Homology, Protein; D013379:Substrate Specificity; D013401:Sugar Alcohol Dehydrogenases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D012255:Ribitol; D013401:Sugar Alcohol Dehydrogenases; C022529:ribitol 2-dehydrogenase
keywords: Enterobacter aerogenes; Homology modeling; Ribitol dehydrogenase; Short-chain dehydrogenase/reductase