The carbazomycin-producing microorganism, strain H 1051-MY 10, was determined to a strain of Streptoverticillium ehimense. Biosynthesis of carbazomycin B was studied using 14C-labeled and 13C-enriched precursors in combination with 13C NMR spectroscopy. The C-2 carbon of [2-13C]trytophan was shown to be involved at the C-3 carbon in carbazomycin B and both carbons of [1,2-13C]acetate at the C-1 and C-10 moiety of the antibiotic. [CH3-13C]Methionine was involved at the methoxyl group but not at the methyl group on the C-2 carbon of the antibiotic. Neither of the labeled carbons, [1-14C]tryptophan nor [2,3-13C]propionic acid, was detected in the antibiotic, and a progenitor of the C-2 and C-11 moiety of the antibiotic has not been determined.