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Published on: January 22, 2014
Gene cloning, expression and characterization of a novel cold-adapted protease from Colwellia hornerae
Xi Xie1, Jiansheng Lin2, Li Lin3
1Guangdong Provincial Key Laboratory of Lingnan Specialty Food Science and Technology, College of Light Industry and Food, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China; Key Laboratory of Green Processing and Intelligent Manufacturing of Lingnan Specialty Food, Ministry of Agriculture, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China.
Abstract:
A cold-adapted protease (Chprotease) from Colwellia hornerae, identified through genomic screening, was cloned, heterologously expressed and purified. The purified Chprotease had a molecular weight of 53.3 kDa and showed maximum specific activity (35.36 U/mL) at 20 °C, with casein as substrate. It was stable and active between the temperature range of 15-25 °C and pH 6.0-8.0, with an optimum temperature of 20 °C at pH 7.0, and it present a good hydrolytic activity to hemoglobin and gluten protein. Moreover, it displayed better tolerance to organic solvents, surfactants, metal ions and reducing agents. Notably, the enzyme exhibited increased activity in the presence of surfactants, showing enhancements of 38 % with 1 % Tween-20 and 13 % with 5 % Tween-80, respectively. Furthermore, it retained over 70 % of its activity in the presence of 5 % SDS, EDTA, β-mercaptoethanol (β-ME), and PMSF. Because of these properties, the Chprotease shows potential for applications in food processing and the production of laundry products.
