Nutrient agar medium with various concentrations of cefotaxime was used for isolation spontaneous mutants from wild type strain of P.aeruginosa PHA-1. Eighty-two mutants were successfully isolated with the viable count 52×107 , these mutants were confirmed as spontaneous not physiological adaption mutants by reculture on the same medium. Then, wild type PHA-1 and mutants were examined for production pyocyanin; a blue greenish pigment was clearly noticed on King A medium. Remarkably the mutant strain named S300-8 was distinguished in productivity in comparison with wild type strain PHA-1; the amount of pigment was 56.0667mg/l and 74.53mg/l respectively. In addition, pyocyanin produced by mutant strain S300-8 revealed a potent efficacy against growth of cancer cell line RD; the low concentration of this pigment caused 65% of dead cells after 72hrs of incubation whereas the cytotoxicity was improved by increasing the concentration of pigment with period of exposure time. Moreover, pyocyanin from mutant strain S300-8 induced apoptosis more than wild type PHA-1; the percentage of DNA fragmentation in RD cell was 80% and 73.3% respectively.
University of Baghdad
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