Abstract: Lysozyme is a natural antibacterial substance that is found in mammalian milk, tears and saliva. The lysozyme content of buffalo（Bubalus bubalis） milk is higher than that of cattle（Bos taurus） milk. There are 16 members of the buffalo lysozyme gene family. This study systematically analyzed the buffalo lysozyme gene family. The results showed that the lysozyme genes of buffalo were closely related to cattle, goat（Capra hircus） and sheep（Ovis aries）. Physicochemical properties of buffalo lysozyme showed that most of the buffalo lysozymes were alkaline, stable, heat-resistant and hydrophilic. Single nucleotide polymorphism analysis showed that the protein encoded by buffalo lysozyme gene had amino acid variation and insertion of amino acid sequence compared with cattle. The variation and insertion of these amino acids may be the reason for the difference in antibacterial ability between buffalo lysozyme and cattle lysozyme. The number of transcription factor binding sites of lysozyme gene in buffalo was higher than that in cattle, which might be the important reason why the expression of lysozyme in buffalo mammary gland was higher than that in cattle.