Abstract
1. Difference spectra of Bacillus megaterium KM membrane preparations indicate the presence of two pigments which bind CO and which exhibit the spectral characteristics of cytochromes a 3 and o. Relative amounts of the pigments vary with growth stage of the organism, but both are present at all stages which have been investigated. The pigments are believed to be metabolically active because they are completely reducible by substrate (NADH) and are reoxidizable in the presence of air. CO difference spectra of whole cell suspensions are in agreement with spectra of the isolated membrane fragments. In particulate preparations and in whole cells, CO difference spectra suggest that the a 3 component binds CO much more readily than the o component; this behavior offers a possible explanation for the fact that cytochrome o has been detected in only a few other microorganisms, since CO binding is by definition the property used to identify this cytochrome. 2. A separation of the two CO-binding pigments is obtained by incubation of membrane preparations with pancreatic lipase. This treatment selectively removes the o pigment from the membrane, leaving the a 3 component associated with an enzymatically active particulate fraction.
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