Abstract: The methylotrophic thermotolerant actinomycete strain 381, isolated from a compost sample, assimilated methanol during growth. Methanol dehydrogenase could not be detected in cell extracts using dye-linked assays in the absence of NAD+. Dichlorophenol-indophenol-linked methanol oxidation was detected in broken cell suspension in the presence of NAD+ and the absence of cyanide. Taxonomic studies showed that actinomycete strain 381 was distinct from Nocardia species 239, which has also been reported to contain an NAD+-dependent methanol dehydrogenase.