Oxygen-insensitive nitroreductases NfsA and NfsB of Escherichia coli function under anaerobic conditions as lawsone-dependent Azo reductases.
Quinones can function as redox mediators in the unspecific anaerobic reduction of azo compounds by various bacterial species. These quinones are enzymatically reduced by the bacteria and the resulting hydroquinones then reduce in a purely chemical redox reaction the azo compounds outside of the cells. Recently, it has been demonstrated that the addition of lawsone (2-hydroxy-1,4-naphthoquinone) to anaerobically incubated cells of Escherichia coli resulted in a pronounced increase in the reduction rates of different sulfonated and polymeric azo compounds. In the present study it was attempted to identify the enzyme system(s) responsible for the reduction of lawsone by E. coli and thus for the lawsone-dependent anaerobic azo reductase activity. An NADH-dependent lawsone reductase activity was found in the cytosolic fraction of the cells. The enzyme was purified by column chromatography and the amino-terminal amino acid sequence of the protein was determined. The sequence obtained was identical to the sequence of an oxygen-insensitive nitroreductase (NfsB) described earlier from this organism. Subsequent biochemical tests with the purified lawsone reductase activity confirmed that the lawsone reductase activity detected was identical with NfsB. In addition it was proven that also a second oxygen-insensitive nitroreductase of E. coli (NfsA) is able to reduce lawsone and thus to function under adequate conditions as quinone-dependent azo reductase.
Rau
Jörg
J
Institut für Mikrobiologie, Universität Stuttgart, Stuttgart, Germany.
Stolz
Andreas
A
D000595:Amino Acid Sequence; D000693:Anaerobiosis; D001391:Azo Compounds / Q000378:metabolism; D024881:Drug Resistance, Bacterial; D019008:Drug Resistance, Neoplasm; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000254:growth & development*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D008969:Molecular Sequence Data; D009285:Naphthoquinones / Q000378:metabolism*; D009601:Nitroreductases / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D010088:Oxidoreductases / Q000378:metabolism; D010100:Oxygen / Q000494:pharmacology*; D013379:Substrate Specificity
D001391:Azo Compounds; D029968:Escherichia coli Proteins; D009285:Naphthoquinones; D010088:Oxidoreductases; C116053:NfsA protein, E coli; C116054:NfsB protein, E coli; D009601:Nitroreductases; D010100:Oxygen; C005090:lawsone