dicumarol and stigmatellin

Previous studies have demonstrated that Shewanella decolorationis S12 can grow on the azo compound amaranth as the sole electron acceptor. Thus, to explore the mechanism of energy generation in this metabolism, membranous vesicles (MVs) were prepared and the mechanism of energy generation investigated. The membrane, which was fragmentized during preparation, automatically formed vesicles ranging from 37.5-112.5 nm in diameter under electron micrograph observation. Energy was conserved when coupling the azoreduction by the MVs of an azo compound or Fe(III) as the sole electron acceptor with H2, formate, or lactate as the electron donor. The amaranth reduction by the vesicles was found to be inhibited by specific respiratory inhibitors, including Cu(2+) ions, dicumarol, stigmatellin, and metyrapone, indicating that the azoreduction was indeed a respiration reaction. This finding was further confirmed by the fact that the ATP synthesis was repressed by the ATPase inhibitor N,N'-dicyclohexylcarbodiimide (DCCD). Therefore, this study offers solid evidence of a mechanism of microbial dissimilatory azoreduction on a subcell level.

Topics: Adenosine Triphosphate; Amaranth Dye; Anti-Bacterial Agents; Cell Membrane; Copper; Dicumarol; Dicyclohexylcarbodiimide; Electron Transport; Enzyme Inhibitors; Ferric Compounds; Isotopes; Metyrapone; Oxidation-Reduction; Polyenes; Reducing Agents; Shewanella; Trace Elements; Uncoupling Agents

Experimental results suggested Shewanella cinica D14T is capable of humus respiration utilizing various organic acids and some important environmental pollutants (e.g., toluene. etc) as electron donors and AQS or AQDS as a sole terminal electron acceptor under anaerobic condition. The dissimilatory reduction of 1mmol/L AQDS can couple to the production of enough ATP to support cell growth about 60 generations; The oxidization of electron donors was coupled to the reduction of humus, as reduced humus increased corresponding with increasing of electron donor; The typical inhibitors such as Cu2+ which inhibited Fe-S center, Stigmatellin which was methyl-naphthoquinone model, Dicumarol which inhibited oxidized methyl-naphthoquinone transform to reduced one, Metyrapone which was specific inhibitor for P450 enzyme blocked the humus respiration seriously. These were powerful evidences for humus-respiration by D14.

Topics: Anaerobiosis; Anthraquinones; Biodegradation, Environmental; Copper; Dicumarol; Electron Transport; Humic Substances; Metyrapone; Oxidation-Reduction; Polyenes; Shewanella