Purification, characterization, and gene cloning of a new cold-adapted beta-galactosidase from Erwinia sp E602 isolated in northeast China

beta-Galactosidases are widely used in industry for elimination of lactose from milk products. A new beta-galactosidase was obtained from bacterial strain Erwinia sp. E602, newly isolated in northeast China. The enzyme was purified with the methods of ammonium sulfate fractionation, ion exchange, and gel filtration chromatography for further study of the enzymatic characteristics. The purified enzyme had a molecular weight of near 110 kDa. The optimum reaction temperature and pH of this enzyme was determined to be 40 degrees C and 7.0, respectively, indicating that this enzyme was a mesophilic neutral beta-galactosidase. Furthermore, the enzyme retained near 10% of the activity at 0 degrees C, which also suggested its cold-adapted property. Kinetics of the beta-galactosidase was studied, and the Km (Michaelis constant) and Vmax (maximum enzymatic reaction rate) of this enzyme were 0.21 mmol/L and 263.16 mu mol/mg per minute, respectively. The effects of metal ions on the enzymatic activity and the lactose hydrolysis efficiency in milk, as well as its trans-glycosylation activity, were studied in this work. The beta-galactosidase coding gene was cloned to be a 3-kb length fragment, which shared at most 81% of identity with the published sequences in NCBI Blast database (https://blast.ncbi.nlm.nih.gov). Results in this work suggested it is a new beta-galactosidase and it has potential to be used in dairy and food processing.