A novel thermostable β-galactosidase from Bacillus coagulans with excellent hydrolysis ability for lactose in whey.

β-Galactosidase is one of the most important enzymes used in dairy industry. Here, a novel thermostable β-galactosidase was cloned and overexpressed from Bacillus coagulans NL01 in Escherichia coli. The phylogenetic trees were constructed using neighbor-joining methods. Phylogeny and amino acid analysis indicated that this enzyme belonged to family 42 of glycoside hydrolases. The optimal pH and temperature were, respectively, 6.0 and 55 to 60°C. The purified enzyme had a 3.5-h half-life at 60°C. Enzyme activity was enhanced by Mn2+. Compared with other β-galactosidases from glycoside hydrolase family 42, B. coagulans β-galactosidase exhibited excellent hydrolysis activity. The Michaelis constant (Km) and maximum rate of enzymatic reaction (Vmax) values for p-nitrophenyl-β-d-galactopyranoside and o-nitrophenyl-β-d-galactopyranoside were 1.06 mM, 19,383.60 U/mg, and 2.73 mM, 5,978.00 U/mg, respectively. More importantly, the enzyme showed lactose hydrolysis ability superior to that of the commercial enzyme. The specific enzyme activity for lactose was 27.18 U/mg. A total of 104.02 g/L lactose in whey was completely hydrolyzed in 3 h with addition of 2.38 mg of pure enzyme per gram of lactose. In view of the high price of commercial β-galactosidase, B. coagulans β-galactosidase could be a promising prototype for development of commercial enzymes aimed at lactose treatment in the dairy industry.