Cell suspensions of yeasts, Candida utilis ATCC 26387, Hansenula polymorpha ATCC 26012, Pichia sp. NRRL-Y-11328, Torulopsis sp. strain A1, and Kloeckera sp. strain A2, grown on various C-1 compounds (methanol, methylamine, methylformate), ethanol, and propylamine catalyzed the oxidation of secondary alcohols to the corresponding methyl ketones. Thus, isopropanol, 2-butanol, 2-pentanol, and 2-hexanol were converted to acetone, 2-butanone, 2-pentanone, and 2-hexanone, respectively. Cell-free extracts derived from methanol-grown yeasts catalyzed an oxidized nicotinamide adenine dinucleotide-dependent oxidation of secondary alcohols to the corresponding methyl ketones, Primary alcohols were not oxidized. The effect of various environmental factors on the production of methyl ketones from secondary alcohols by methanol-grown Pichia sp. was investigated.