1887

Abstract

The ability to use sulphite as a respiratory electron donor is usually associated with free-living chemolithotrophic sulphur-oxidizing bacteria. However, this paper shows that the chemoheterotrophic human pathogen has the ability to respire sulphite, with oxygen uptake rates of 23±8 and 28±15 nmol O min (mg cell protein) after the addition of 0·5 mM sodium sulphite or metabisulphite, respectively, to intact cells. The NCTC 11168 and genes encode a monohaem cytochrome and molybdopterin oxidoreductase, respectively, homologous to the sulphite : cytochrome oxidoreductase (SOR) of . Western blots of periplasm probed with a SorA antibody demonstrated cross-reaction of a 45 kDa band, consistent with the size of Cj0005. The gene was inactivated by insertion of a kanamycin-resistance cassette. The resulting mutant showed wild-type rates of formate-dependent respiration but was unable to respire with sulphite or metabisulphite as electron donors. 2-Heptyl-4-hydroxyquinoline--oxide (HQNO), a cytochrome complex inhibitor, did not affect sulphite respiration at concentrations up to 25 μM, whereas formate respiration (which occurs partly via a dependent route) was inhibited 50 %, thus suggesting that electrons from sulphite enter the respiratory chain after the complex at the level of cytochrome . Periplasmic extracts of wild-type 11168 showed a symmetrical absorption peak at 552 nm after the addition of sulphite, demonstrating the reduction of cytochrome . No cytochrome reduction was observed after addition of sulphite to periplasmic extracts of the mutant. A fractionation study confirmed that the majority of the SOR activity is located in the periplasm in , and this activity was partially purified by ion-exchange chromatography. The presence of a sulphite respiration system in is another example of the surprising diversity of the electron-transport chain in this small-genome pathogen. Sulphite respiration may be of importance for survival in environmental microaerobic niches and some foods, and may also provide a detoxification mechanism for this normally growth-inhibitory compound.

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2005-01-01
2024-03-28
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