Document Type : Article
Authors
1
Department of Medical Genetics, National Institute of Genetic Engineering and Biotechnology (NIGEB)
2
Bioprocess Engineering Research Group, Departments of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB)
3
Faculty of Biotechnology, Amol University of Special Modern Technologies, Amol, Iran, and Departments of Biochemistry, Institute of Biochemistry and Biophysics (IBB), University of Tehran, Tehran, Iran
4
National institute of genetic Engineering and Biotechnology Shahrak-e pajoohesh,km 15,Tehran-karaj Highway, Tehran,Iran
5
Department of Biochemistry, Faculty of Biological Science, Tarbiat Modares University, Tehran, Iran.
Abstract
Abstract
Background: Chitinase is one of the essential enzymes that have broad applications in industrial, agriculture, medical through biodegradation of chitin and hence are in demand. Chitinase-producing Serratia marcescens B4A was cultured in the medium containing chitin and then was purified through ammonium sulfate precipitation and DEAE ion-exchange chromatography. The purity of chitinase was determined by one and two- dimension sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
Results: Its optimum activity was at pH 7.0 and temperature 50 °C and was stable at 90 °C for 60 min and over a pH range from 5.0 to 8.0. The Km, Vmax, kcat, and kcat/Km values were 3.72 mg/ml, 0.19 U/ml, 134.75 min-1, and 36.17 mg.min-1.ml-1, respectively that showed a high affinity of chitinase to the substrate and exhibited excellent catalytic efficiency. Investigation of irreversible thermo-inactivation of chitinase at a range of 60 to 90 °C revealed a high value of ΔG# with the low value of ΔH# and a negative value of ΔS#.
Conclusions: These characteristics of Serratia marcescens B4A chitinase showed high tolerance to thermal denaturation. Therefore, it may have a positive impact on industrial, antifungal applications, and biodegrade chitin waste. The present study is the first report on the thermodynamic and kinetic characterization of chitinase from Serratia marcescens B4A.
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