PRODUCTION, OPTIMIZATION AND PARTIAL PURIFICATION OF NATTOKINASE FROM BACILLUS CEREUS; A STRAIN ISOLATED FROM GRAPE WINE

Abstract

In the current study a potent fibrinolytic protease producing Bacillus cereus VMI2 was isolated from Grape wine (Vitis vinifera) and studied the protease activity and fibrinolytic activity. The organism was identified by morphological, biochemical and molecular analysis. The potent Bacillus cereus VMI2 showed significant clot lysis activity in 2h of incubation. In order to enhance the fibrinolytic activity, the medium was optimized. Inoculum size, pH, temperature, nitrogen and carbon were selected for the optimization. Compared with base medium the optimized medium with fructose, beef extract and 1000 µL inoculum volume were incubated at 35℃ and showed significant increase in fibrinolytic activity. The potent strain also showed a maximum of enzyme production and enzyme activity at 6^th h which showed the optimum time for production of nattokinase. The FTIR-HPLC results confirmed the presence of nattokinase. The molecular weight of the enzyme was analysed by SDS PAGE. It was found to be a 29kDa protein, confirmed the presence of nattokinase.