Isolation, Purification and Characterization of Bacitracin from Bacillus

Bacterial cultures of bacillus species capable of producing bacitracin were isolated from local habitats (garden soil, timber yard soil and sea water) and screened for the production of bacitracin on nutrient media against test organisms viz: Micrococcus luteus,(ATCC 9341) & Staphylococcus aureus (ATCC 13565).Optimization of culture conditions and media composition for bacitracin production was done by investigating the effect of different nitrogen sources ( tryptone, meat extract, yeast extract ), carbon sources (maltose, glucose, fructose, sucrose, lactose) pH (3.6, 4.6, 5.6, 6.6), time(6hrs,12hrs,16hrs,24hrs,48hrs 72hrs), temperature (4°C,15°C,25°C,37°C,48°C ) on the growth of isolates. The isolated strains were subjected to solid state and submerged fermentation on different production media containing Soyabean meal & Wheat bran. The bacitracin was extracted by butanol-ether solvent system and purified by thin layer chromatography and characterized by SDS-PAGE. Keyword: Bacitracin, SDS-PAGE, Antimicrobial activity