Isolation and Characterization of Buffalo Milk Lysozyme

Lysozyme is a commercially valuable enzyme, and is applied in many fields, concerning products such as foods, drugs, and the like. In this work, lysozyme was isolated and purified from buffalo milk using sephadex G-50 and cation exchanger carboxymethyl cellulose. Lysozyme active fractions from buffalo milk were assayed against Gram positive substrate Micrococcus luteus at 450 nm and a decline in absorbance of 0.001 per min was observed. The optimum activity of lysozyme (158.3 ± 1.7 units/mL) was at 7.5 pH and 37°C temperature. Lysozyme activity at pasteurization temperatures 62.5°C, 30 min and 75°C, 15 s were (156.08 ± 1.03 and 156 ± 2 units/mL) not affected significantly; however, 47% activity of lysozyme was reduced at 100°C for 5 min. Antibacterial susceptibility testing of lysozyme (chicken egg white lysozyme and buffalo milk lysozyme) was performed on Micrococcus luteus (ATCC 4698) and Escherichia coli (ATCC 25235). Both lysozymes showed no inhibition effect against Escherichia coli.