The cytochrome composition of the meat spoilage bacterium Brochothrix thermosphacta: identification of cytochrome a3-and d-type terminal oxidases under various conditions.

Abstract

Brochothrix thermosphacta, grown in batch culture in a yeast-dextrose broth, at temperatures from 30 degrees C to 10 degrees C, contained diverse membrane-bound respiratory cytochromes. Under conditions of moderate aeration, cytochromes of the a-, b- and d-type were detected at all growth temperatures, but the proportions changed as a function of temperature, with the spectra of cells grown at 10 or 15 degrees C being dominated by a-type cytochrome(s). Cytochrome a3 was detected by its reactions with CO and cyanide in cells from all growth conditions. An additional cytochrome a, which was not cyanide-reactive, was also detected, suggesting the presence of an aa3 oxidase complex. Cytochrome d was cyanide- and CO-reactive, but not detectable in photodissociation spectra, presumably because of the very rapid recombination of CO at the sub-zero temperatures used. Decreasing the oxygen transfer rates to batch cultures resulted in enhanced expression of cytochrome d and changed the proportion of the aa3-type oxidase that could be attributed to ligand-binding cytochrome a3; at the lowest oxygen transfer rates, no cytochrome a was detected, suggesting the presence of a cytochrome ba3 terminal oxidase complex. Intact cells showed no evidence of a c-type cytochrome and no haem C was detected in membrane preparations. After growth at 10 degrees C, the cytochrome composition of B. campestris was essentially identical to that of B. thermosphacta. The multiplicity of putative terminal oxidases in B. thermosphacta is discussed.