Purification, characterization, antibacterial activity and N-terminal sequencing of buffalo-milk lysozyme.

摘要: Lysozyme from buffalo milk was purified to homogeneity and its N-terminal amino acid sequence, biochemical properties antibacterial spectrum were determined. The purification procedure, comprising ion-exchange chromatography using CM-cellulose size-exclusion Sephadex G-50, conferred 8622-fold 39.3% recovery of lyoszyme. enzyme migrated as a single band on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) native PAGE. Immunological purity lysozyme preparation confirmed by immuno-electrophoresis. Molecular weight buffalo-milk determined SDS-PAGE 16 kDa composition reverse phase high performance liquid (HPLC). sequence 23 residues at the end showed 56.5% homology with bovine 30.4% equine lysozyme. specific activity ten-times that Buffalo-milk active over wide range pH strongly influenced molarity medium. Antibacterial against 11 species bacteria; out seven Gram-positive bacteria tested, four inhibited, while Gram-negative resistant.