Purification and characterization of beta-galactosidase from a strain of Bacillus coagulans

Abstract

beta-Galactosidase from B. coagulans strain L4 is produced constitutively, has a mol. wt. of 4.3 x 10(5) and an optimal temperature of 55 degrees C. The optimal pH at 30 degrees C is 6.0 whereas at 55 degrees C it is 6.5. The energy of activation of enzyme activity is 41.9 kJ/mol (10 kcal/mol). No cations are required. The Km with ONPG as substrate is 4.2-5.6 mM and with lactose is 50 mM. The Ki for inhibition by galactose is 11.7-13.4 mM and for dextrose is 50 mM. Galactose inhibited competitively while dextrose inhibited noncompetitively. The purified and unprotected enzyme is 70% destroyed in 30 min at 55 degrees C whereas in the presence of 2 mg/ml of BSA 42% of the activity is destroyed in 30 min at 55 degrees C. An overall purification of 75.3-fold was achieved.