Partial purification and characterization of protease enzyme from Bacillus subtilis and Bacillus cereus.
The aim of this experimental study was to isolate and partially purify protease enzyme from Bacillus cereus and Bacillus subtilis. Protease enzyme is obtained by inducing spore genesis of bacteria from Bacillus species in suitable nutrient plates. The partial purification was realized by applying, respectively, ammonium sulfate precipitation, dialysis, and DEAE-cellulose ion-exchange chromatography to the supernatant that was produced later. Optimum pH, optimum temperature, pH stability, and temperature stability were determined, as well as the effects of pH, temperature, substrate concentration, reaction time, and inhibitors and activators on enzyme activity. In addition, the molecular mass of the obtained enzyme was investigated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The specific activity of partially purified enzyme from B. subtilis was determined to be 84 U/mg. The final enzyme preparation was eight-fold more pure than the crude homogenate. The molecular mass of the partially purified enzyme was found to be 45 kDa by using SDS-PAGE. The protease enzyme that was partially purified from B. cereus was purified 1.2-fold after ammonium sulfate precipitation. The molecular mass of the partially purified enzyme was determined to be 37 kDa by using SDS-PAGE.
Orhan
Elif
E
Department of Chemical Engineering, Istanbul Technical University, 34469 Maslak, Istanbul, Turkey.
Omay
Didem
D
Güvenilir
Yüksel
Y
D001409:Bacillus cereus / Q000201:enzymology*; D001412:Bacillus subtilis / Q000201:enzymology*; D018929:Cell Culture Techniques / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D003956:Dialysis / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004789:Enzyme Activation; D004795:Enzyme Stability; D005590:Fractional Precipitation; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000737:chemistry* / Q000302:isolation & purification*; D013696:Temperature
D010447:Peptide Hydrolases