Calcium dependent, alkaline detergent-stable trypsin from the viscera of Goby (Zosterisessor ophiocephalus): Purification and characterization
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- 作者：Rim Nasri ; Assaâ ; d Sila ; Naourez Ktari ; Imen Lassoued ; Ali Bougatef ; Maha Karra-Chaâ ; bouni ; Moncef Nasri ; moncef.nasri@enis.rnu.tn ; mon_nasri@yahoo.fr
- 关键词：Alkaline trypsin ; Biochemical characterization ; Goby ; Zosterissessor ophiocephalus ; Calcium-dependent ; Detergent-stable
- 刊名：Process Biochemistry
- 出版年：2012
- 出版时间：December, 2012
- 年：2012
- 卷：47
- 期：12
- 页码：1957-1964
- 全文大小：694 K
文摘
An alkaline calcium dependent trypsin from the viscera of Goby (Zosterisessor ophiocephalus) was purified to homogeneity with a 16-fold increase in specific activity and 20 % recovery. The purified trypsin appeared as a single band on sodium dodecyl sulphate-polyacrylamide gel (SDS-PAGE) and native-PAGE. The enzyme had an estimated molecular weight of 23.2 kDa.
The optimum pH was 9.0, and the enzyme was extremely stable in various pH buffers between pH 7.0 and 11.0. The optimum temperature for enzyme activity was 60 ¡ãC, and the activity and stability of trypsin was highly dependent on the presence of calcium ion. At 60 ¡ãC, Ca2+ (5 mM) stimulated the protease activity by 220 % . The trypsin kinetic constants, Km and kcat, were 0.312 mM and 2.03 s?1.
The enzyme showed high stability towards non-ionic surfactants and oxidizing agent. In addition, the enzyme showed excellent stability and compatibility with some commercial solid and liquid detergents.