title: Role of hydrogenase 1 of Clostridium pasteurianum in the reduction of metronidazole.
abstract: Competition studies between the phosphoroclastic reaction and the metronidazole reduction reaction using dialyzed crude cell-free extracts of Clostridium pasteurianum which were essentially devoid of Hydrogenase 1 activity demonstrated that this enzyme plays an important role in the reduction of metronidazole. To determine further the exact function for Hydrogenase 1 in the reduction of the drug, this enzyme was highly purified from C. pasteurianum. Metronidazole reduction activity copurified with Hydrogenase 1 specific activity throughout the purification procedure. Drug reduction required the presence of an electron carrier and could not be accomplished by the enzyme alone. Ferredoxin, and also the low potential electron carrier dyes, methyl and benzyl viologen, and the flavin coenzymes, FAD and flavin mononucleotide (FMN), could couple the reduction of metronidazole. Hydrogenase 1 activity and its metronidazole reduction activity were inactivated irreversibly in the presence of oxygen. Metronidazole could be reduced only by an electron carrier-Hydrogenase 1 mechanism or directly by sodium dithionite.
authors.affiliation: Department of Microbiology and Infectious Diseases, University of Calgary, Alberta, Canada.
authors.forename: D L
authors.initials: DL
authors.lastname: Church
authors.forename: H R
authors.initials: HR
authors.lastname: Rabin
authors.forename: E J
authors.initials: EJ
authors.lastname: Laishley
mesh_terms: D003013:Clostridium / Q000201:enzymology*; D004227:Dithionite; D004579:Electron Transport; D006864:Hydrogenase / Q000302:isolation & purification / Q000378:metabolism*; D008795:Metronidazole / Q000378:metabolism*; D009243:NAD; D009249:NADP; D010084:Oxidation-Reduction; D010269:Paraquat
chemical_list: D009243:NAD; D008795:Metronidazole; D004227:Dithionite; D009249:NADP; D006864:Hydrogenase; D010269:Paraquat