Gene cloning, expression and characterization of a novel cold-adapted protease from Planococcus sp.

Publication date: Available online 30 April 2016 Source:Journal of Molecular Catalysis B: Enzymatic Author(s): Huitu Zhang, Huiyan Mu, Qingshan Mo, Tongwei Sun, Yang Liu, Mingyu Xu, Haikuan Wang, Yujie Dai, Fuping Lu A psychrophilic bacterium strain named M7 was isolated from the deep-sea mud of Eastern Indian Ocean and identified as Planococcus sp. A novel protease gene, cpls8, was cloned from the newly isolated bacterium and successfully expressed in Escherchia coli. The open reading frame of cpls8 gene encodes a 329-amino acid polypeptide with a molecular weight of approximately 35.6kDa. Sequence alignment reveals that the CPLS8 protease contain a catalytic module belonging to serralysin-type protease family 8 and displays the highest identity of 62% to a functionally known protease from Bacillus sp. The heterologously expressed CPLS8 exhibited optimal activity at pH 10.0 and demonstrated an increasing activity at temperatures between 5 and 35°C with a complete inactivation at 55°C. At 25°C the relative activity of CPLS8 still reached to 80% of the highest activity at 35°C. These enzymatic properties together with amino acid composition analysis showed that CPLS8 was a novel cold-adapted protease and seemed to be attractive for industrial application. Graphical abstract
Source: Journal of Molecular Catalysis B: Enzymatic - Category: Biochemistry Source Type: research