A novel transglycosylating beta-galactosidase was purified from Enterobacter agglomerans B1. It was a homodimer of approximately 248 kDa. The optimal pH and temperature for oNPGal hydrolysis were 7.5-8.0 and 37-40 degrees C, respectively. The K(m) values for oNPGal and lactose were 0.06 and 114 mM,