Isolasi, Pemurnian Parsial, dan Karakterisasi Enzim Kitinase dari Bacillus sp. BPPT CC 2

Abstract

Chitinase is a potential enzyme due to its acitvity hydrolyting chitin to its derivates. Therefore, it can be used for various applications, such as agriculture, microbiology, and cosmetic. This study aimed to isolate, purify, and characterize (pH, temperature, metal ion effect, molecular weight, and kinetic parameters) chitinase of Bacillus sp. BPPT CC 2. Ammonium sulphate precipitation and dialysis were used for enzyme purification. Enzyme was characaterized its optimum pH and temperature, metal ions, molecular weight, as well as kinetic parameters. The results showed that enzyme was well precipitated using 80% ammonium sulphate. After dialyzed, level of enzyme purity increased 6.58 fold with specific activity of 0.0924 U/mg. Results showed that the optimum temperature and pH of chitinase were found at 55ºC and pH 5. Value of Km and Vmaks values were 2.705x10-4mg/L second and 0.6959 mg/L. Predicted molecular weight of chitinase was 103 kDa. Addition of metal ions Mn2+and EDTA (5 mM and 10 mM) act as activators of chitinase