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The mitomycins were first obtained in 1956 from Streptomyces caespitosus by Hata and coworkers in Japan (1, 2). The isolation involved alumina chromatography of the chloroform extracts of the concentrated aqueous filtrates of the fermentation broth. Although there were many active components in the eluted fractions, only two antibiotics, mitomycins A and B were obtained in crystalline form. Later fractionations of broths of S. caespitosus yielded mitomycin C (3). By 1958, the outstanding antitumor activity of mitomycin C was a subject of great interest in spite of the fact that there existed no firm evidence as to its structure (4). In 1960, N-methylmitomycin C, called porfiromycin, was isolated from cultures of Streptoverticillatium ardus (5). In 1962 the isolation of the same four antibiotics from Streptomyces verticillatus was reported in the U.S. (6). Additionally, a fifth, but inactive member of mitomycin family, mitiromycin was obtained. The fractionation of the S. verticillatus broths to afford the crystalline antibiotics was accomplished by partition chromatography on a diatomaceous earth support.