Isolation, purification and mass production of protease enzyme from bacillus subtilis

Screening and isolation of protease producing strains of bacteria were carried out from four different soil samples collected from various places in Bangalore. The isolates were positive on skim milk agar (1%) and thus are selected as protease producing strain. The organisms were tested for various biochemical tests, which lead to their identification as Bacillus subtilis producing protease enzyme. These Bacillus subtilis could group upto 40oC and pH range 6-9 with optimal growth temperature and pH at 37oC and 8.0 respectively. It was also optimized for carbon test and nitrogen test with optimal growth in dextrose and peptone respectively. Enzyme production was carried in 1 litre of optimized media in the fermenter at 37oC for 48 hours at pH 8.0. Harvested protease product was purified by salt precipitation method. Finally the enzyme protease was purified by column chromatography. The protein was characterized using SDS-PAGE. This result showed that Bacillus subtilis under study is a good producer of extra cellular protease, which can be beneficial for industries.