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The effect of adding hemin to anaerobically grown cells of a strain of Staphylococcus epidermidis, which was heme-deficient due to anaerobic growth, has been examined. Cells grown anaerobically in media containing hemin exhibited a marked increase in several oxidative activities as compared with cells grown anaerobically without hemin. The respiratory activity of whole cells and a cyamide-sensitive reduced nicotinamide adenine dinucleotide oxidase activity of cell-free extracts were increased fourfold. The content of enzymatically reducible pigments which exhibit difference spectra similar to cytochromes b(1) and o was also markedly increased. These pigments are mostly sedimented at 100,000 x g (1 hr). Hemin also caused a marked increase in respiratory activity when added directly to the anaerobic culture after the period of growth, but did not cause a similar increase in respiration when added to washed, resting-cell suspensions. Under the latter conditions, heme pigments were formed which exhibited difference spectra similar to, but not identical with, the spectra of pigments found in anaerobic cells grown in the presence of hemin. When resting suspensions of cells grown anaerobically without hemin were exposed to air, a rapid fourfold increase in respiratory activity and a limited increase in cytochrome-like pigments occurred. The presence of the heme precursor Delta-aminolevulinic acid during this aeration resulted in a rapid and marked increase in heme pigments, but only a slight stimulation of respiratory activity. The possible implications of these results for the roles which heme and oxygen play in the development of the respiratory system of this organism are discussed.

Effects of Oxygen and Heme on the Development of a Microbial Respiratory System