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Mitomycin C (MC) is an antitumor antibiotic derived biosynthetically from 3-amino-5-hydroxybenzoic acid (AHBA),d -glucosamine, and carbamoyl phosphate. A gene (mitA ) involved in synthesis of AHBA has been identified and found to be linked to the MC resistance locus,mrd , inStreptomyces lavendulae . Nucleotide sequence analysis showed thatmitA encodes a 388-amino-acid protein that has 71% identity (80% similarity) with the rifamycin AHBA synthase fromAmycolatopsis mediterranei , as well as with two additional AHBA synthases from related ansamycin antibiotic-producing microorganisms. Gene disruption and site-directed mutagenesis of theS. lavendulae chromosomal copy ofmitA completely blocked the production of MC. The function ofmitA was confirmed by complementation of anS. lavendulae strain containing a K191A mutation in MitA with AHBA. A second gene (mitB ) encoding a 272-amino-acid protein (related to a group of glycosyltransferases) was identified immediately downstream ofmitA that upon disruption resulted in abrogation of MC synthesis. This work has localized a cluster of key genes that mediate assembly of the unique mitosane class of natural products.

journal of bacteriology

Genetic Localization and Molecular Characterization of Two Key Genes ( <i>mitAB</i> ) Required for Biosynthesis of the Antitumor Antibiotic Mitomycin C

Genetic Localization and Molecular Characterization of Two Key Genes ( mitAB ) Required for Biosynthesis of the Antitumor Antibiotic Mitomycin C