Rubradirin, as ansamycin antibiotic has been isolated from streptomyces achromogenes var rubradiris NRRL 3061. It Consists of four distinct structural moieties. Amide bond present in the structure of rubradirin plays an important role as a linkage between the coumarin ring and didipicolinic acid moiety of the rubradirin. We disrupted the amide bond present in the structure of rubradirin and then analyzed the production of rubradirin could not take place in mutant. The rubransarol containing fractions and the native rubradirin were detected by HPLC.