Abstract:
Protease enzyme from Bacillus cereus LS2B was successively purified by four step
processes including centrifugation, ammonium precipitation, membrane dialysis,
and hitrap ion exchange with DEAE sepharose FF matrix. The best concentration
of ammonium sulfate to purified enzyme was observed 80%. Hitrap ion exchange
chromatography was performed with flow rate 1.5 ml min-1, and the number of
enzyme in once running was 52,5 ml in the 35 tube. Specific activities at crude enzyme,
ammonium sulfate precipitation, membrane dialysis and hitrap ion exchange was 0.4
U/mg, 0.5 U/ml, 1.8 U/mg and 7.2 U/mg, respectively. By using hitrap ion exchange
purification of alkaline protease from Bacillus cereus LS2B was observed 16 fold from
crude enzyme. Furthermore, the yield decreased from 100% in crude enzyme to 2% by
using hitrap ion exchange purification. The purified enzyme was characterized using
SDS-PAGE obtained three specific protein molecules with each weight was 34 kDa,
17 kDa and 13 kDa respectively. The maksimum activity of pure anzyme alkaline
protease from Bacillus cereus LS2B obtained at 40o C and pH 8.
