Cloning, expression, and bioinformatics analysis and characterization of a β-galactosidase from Bacillus coagulans T242

Abstract

The activities of β-galactosidases from bacteria and molds are affected by temperature, pH, and other factors in the processing of dairy products, limiting their application, so it is necessary to find alternative lactases. In this study, the β-galactosidase gene from Bacillus coagulans T242 was cloned, co-expressed with a molecular chaperone in Escherichia coli BL21, and subjected to bioinformatic and kinetic analyses and lactase characterization. The results show that the enzyme is a novel thermostable neutral lactase with optimum hydrolytic activity at pH 6.8 and 50°C. The thermal stability and increased lactose hydrolysis activity of β-galactosidase in the presence of Ca²⁺ indicated its potential application in the dairy industry.

Keywords: Bacillus coagulans; bioinformatics analysis; enzymatic properties; expression; thermostable β-galactosidase.