Spores of Septomyxa affinis transformed Reichstein"s compound "S" into mainly 1-dehydro "S" and small amounts of 17-side-chain degradation products. 1-Dehydrogenating activity of the spores was not influenced by the sporulation medium. Adaptation has never been observed. Metal ions, chelating agents, activators, and cofactors showed no significant influence, within wide limits. Externally supplied phosphate, glucose, or any other carbon source was not required. Mercuric chloride, N-ethylmaleimide, NaN3, 2,4-dinitrophenol, antimycin, and some oxidation–reduction dyes inhibited steroid conversion. Degradation of the side chain was blocked by conducting the transformation at 32 °C.Spores of S. affinis oxidized acetate, glucose, fructose, ethanol, and xylose. Oxidation of these substrates was inhibited by most of the reagents that also inhibited steroid transformation.