title: Novel dehalogenase mechanism for 2,3-dichloro-1-propanol utilization in Pseudomonas putida strain MC4.
abstract: A Pseudomonas putida strain (MC4) that can utilize 2,3-dichloro-1-propanol (DCP) and several aliphatic haloacids and haloalcohols as sole carbon and energy source for growth was isolated from contaminated soil. Degradation of DCP was found to start with oxidation and concomitant dehalogenation catalyzed by a 72-kDa monomeric protein (DppA) that was isolated from cell lysate. The dppA gene was cloned from a cosmid library and appeared to encode a protein equipped with a signal peptide and that possessed high similarity to quinohemoprotein alcohol dehydrogenases (ADHs), particularly ADH IIB and ADH IIG from Pseudomonas putida HK. This novel dehalogenating dehydrogenase has a broad substrate range, encompassing a number of nonhalogenated alcohols and haloalcohols. With DCP, DppA exhibited a k(cat) of 17 s(-1). (1)H nuclear magnetic resonance experiments indicated that DCP oxidation by DppA in the presence of 2,6-dichlorophenolindophenol (DCPIP) and potassium ferricyanide [K(3)Fe(CN)(6)] yielded 2-chloroacrolein, which was oxidized to 2-chloroacrylic acid.
authors: Department of Biochemistry, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, Netherlands.
authors: Muhammad Irfan
authors: MI
authors: Arif
authors: Ghufrana
authors: G
authors: Samin
authors: Jan G E
authors: JG
authors: van Leeuwen
authors: Jantien
authors: J
authors: Oppentocht
authors: Dick B
authors: DB
authors: Janssen
mesh_terms: D000171:Acrolein / Q000031:analogs & derivatives / Q000378:metabolism; D002244:Carbon / Q000378:metabolism; D002728:Chlorohydrins / Q000378:metabolism*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004734:Energy Metabolism; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D021382:Protein Sorting Signals; D016958:Pseudomonas putida / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D002728:Chlorohydrins; D004269:DNA, Bacterial; D021382:Protein Sorting Signals; C028406:2-chloroacrolein; D002244:Carbon; D000171:Acrolein; C076159:2,3-dichloro-1-propanol; D010088:Oxidoreductases