Hydrogenase activity and the H2-fumarate electron transport system in Bacteroides fragilis.
Hydrogenase activity and the H(2)-fumarate electron transport system in a carbohydrate-fermenting obligate anaerobe, Bacteroides fragilis, were investigated. In both whole cells and cell extracts, hydrogenase activity was demonstrated with methylene blue, benzyl viologen, flavin mononucleotide, or flavin adenine dinucleotide as the electron acceptor. A catalytic quantity of benzyl viologen or ferredoxin from Clostridium pasteurianum was required to reduce nicotinamide adenine dinucleotide or nicotinamide adenine dinucleotide phosphate with H(2). Much of the hydrogenase activity appeared to be associated with the soluble fraction of the cell. Fumarate reduction to succinate by H(2) was demonstrable in cell extracts only in the presence of a catalytic quantity of benzyl viologen, flavin mononucleotide, flavin adenine dinucleotide, or ferredoxin from C. pasteurianum. Sulfhydryl compounds were not required for fumarate reduction by H(2), but mercaptoethanol and dithiothreitol appeared to stimulate this activity by 59 and 61%, respectively. Inhibition of fumarate reduction by acriflavin, rotenone, 2-heptyl-4-hydroxyquinoline-N-oxide, and antimycin A suggest the involvement of a flavoprotein, a quinone, and cytochrome b in the reduction of fumarate to succinate. The involvement of a quinone in fumarate reduction is also apparent from the inhibition of fumarate reduction by H(2) when cell extracts were irradiated with ultraviolet light. Based on the evidence obtained, a possible scheme for the flow of electrons from H(2) to fumarate in B. fragilis is proposed.
D000167:Acriflavine / Q000494:pharmacology; D000693:Anaerobiosis; D000968:Antimycin A / Q000494:pharmacology; D001441:Bacteroides fragilis / Q000201:enzymology / Q000378:metabolism*; D004579:Electron Transport / Q000187:drug effects / Q000528:radiation effects; D005650:Fumarates / Q000378:metabolism*; D006859:Hydrogen / Q000378:metabolism; D006912:Hydroxyquinolines / Q000494:pharmacology; D010088:Oxidoreductases / Q000378:metabolism*; D012402:Rotenone / Q000494:pharmacology; D013438:Sulfhydryl Compounds / Q000494:pharmacology; D014466:Ultraviolet Rays; D014812:Vitamin K / Q000494:pharmacology
D005650:Fumarates; D006912:Hydroxyquinolines; D013438:Sulfhydryl Compounds; D012402:Rotenone; D014812:Vitamin K; D000167:Acriflavine; D000968:Antimycin A; D006859:Hydrogen; D010088:Oxidoreductases