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The enzyme -galactosidase has tremendous potential in research and application in various fields like food, bioremediation, biosensor, diagnosis and treatment of disorders. The sources of the -galactosidase are microorganisms, plants and animals. -galactosidase producing bacteria was isolated from milk and milk products. Bacteria were tested for their ability to hydrolyze 5- bromo-4-chloro-3-indolyl- -D-galactosidase (X-Gal) and O-nitro phenyl- -D-galactopyranoside (ONPG). Further, it was characterized by biochemical and molecular methods.