Production of Protease from Bacillus sp.isolated from Natural Habitats

Alkaline protease producing Baciilus spp. were isolated from natural habitats of Lahore. Thirty eight isolates exhibited the prominent zones of clearance on skim milk agar medium at pH 10. These isolates which showed best proteolytic activity were then identified. Twenty two (60 %) of the alkalophilic isolates belonged to the genus Bacillus. Maximum enzyme activity (191.0U/ ml) was found in strain isolated from slaughterhouse (coded as MPPL1) at pH 10 and temperature 37°C. Maximum activity was obtained at pH 10, 140 rpm, at 37°C after 24 hours of incubation. The results indicated that protease producing acillus strain coded as MPPL1 is active at normal ranges of temperature and high pH. Alkaline protease producing conditions i.e, culture media, pH, temperature, agitation speed, incubation period were optimized at shake flask level for maximum enzyme activity and these conditions were applied at bioreactor for submerged fermentation in 1 litre jar bioreactor, an increase of nearly 80% proteolytic activity was observed after obtaining crude enzyme by centrifugation the fermented broth at 10000rpm for 10 minutes at 4°C.