Poster Presentation 1B-20

Partial Purification and Characterization of Protease from Bacillus subtilis and Bacillus cereus  Bacteria

 

 

Elif Orhan, Didem Omay and Yüksel Güvenilir*

 

 

Istanbul Technical University

Department of Chemical Engineering

80626, Maslak, Istanbul, TURKEY

Phone:  +90 212 2656294

E-mail: avcibasi@itu.edu.tr

 

 

 

The aim of this experimental study is partial isolation and purification of protease enzyme from Bacillus cereus and Bacillus subtilis bacterial species. Protease enzyme is obtained by producing spore genesis of bacteria, which is from Bacillus species in suitable food places.

 

The purification was realized by applying respectively ammonium sulphate precipitation, dialysis, DEAE-Cellulose ion exchange chromatography to the supernatant that was produced later. In this study, the effect of substrate concentration, reaction time, the effect of inhibitor and activator on the optimum pH, optimum temperature, pH stability, temperature stability was determined. Besides, molecular weight of the obtained enzyme was investigated by realization of transaction of SDS-PAGE.

 

In this study, specific activity of partial purified enzyme from Bacillus subtilis species was found as 84 U/mg. The final enzyme preparation was 8 folds purer than the crude homogenate. Molecular weight of the purified enzyme was determined. It was found that the weight of enzyme was 45 kDa by using SDS-PAGE.

 

Protease enzyme which was purificated from Bacillus cereus bacteria species was purified 1.2 fold after ammonium sulphate precipitation. Molecular weight of the purified enzyme was determined 37 kDa by using SDS-PAGE.