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Received January 8, 2007
Accepted February 27, 2007
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Purification and characterization of a thermophilic chitinase produced by Aeromonas sp. DYU-Too7
Department of Bioindustry Technology, Da-Yeh University, Chang-hua 515, Taiwan 1Department of Environmental Engineering, Da-Yeh University, Chang-hua 515, Taiwan
Korean Journal of Chemical Engineering, September 2007, 24(5), 806-811(6), 10.1007/s11814-007-0045-3
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Abstract
An extracellular chitinase, produced by Aeromonas sp. DYU-Too7, was purified in the following procedures: ammonium sulfate precipitation, ultrafiltration, chromatographic separation of DEAE-sepharose CL-6B and sephacryl S-100HR. The resulting chitinase has a molecular mass of 36 kDa, an optimal reaction pH of 5.0, and an optimal reaction temperature of 70 ℃. It retains almost 100% activity in the pH range of 5.0-8.0. This chitinase has a high thermal tolerance and retained 90% of its activity at 50 ℃ and 75% at 60 ℃. Enzyme activity was inhibited by Ba2+, Hg2+, Mg2+ and Ag+ cations, but was not substantially inhibited by the K+ cation nor the chelating agent EDTA. The Km and Vmax, using colloidal chitin as a substrate, are 6.3 g/L and 18.69 μmol/min/mg-protein, respectively. The 36 kDa chitinase of Aeromonas sp. DYU-Too7 is an exo-type enzyme, because chitobiose was the main hydrolysate in hydrolysis of colloidal chitin.
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References
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Wen CM, Tseng CS, Cheng CY, Li YK, Biotechnol. Appl. Biochem., 35, 213 (2002)
