ISOLATION OF HEN EGG WHITE LYSOZYME BY CATION EXCHANGE CHROMATOGRAPHY, ANALYSIS OF ITS DIGESTIBILITY AND EVALUATION OF THE INHIBITION LIPID PEROXIDATION IN THE ZEBRAFISH MODEL
Objective: The aim of this study was to separate and identify lysozyme using cation exchange chromatography, evaluate the protein digestibility and analyze the inhibition lipid peroxidation in the zebrafish model. Methods: Hen egg white lysozyme was isolate with cation exchange chromatography. Residual muramidase activity was evaluated with the spectrophotometric method. Isolate lysozyme (ILZ) and hydrolysates were analyzed with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) electrophoresis and reversed phase high performance liquid chromatography (RP HPLC) Results: To identify the protein, the sample was isolated with cationic exchange chromatography, sample was analyzed using RP HPLC and SDS PAGE confirming that the fraction was hen lysozyme egg white. The enzymatic activity of the isolated protein was normal compared to the commercial lysozyme activity. Hydrolysates had no muramidase activity and were able to inhibit lipid peroxidation in zebrafish larvae. Conclusions: Cation exchange chromatography is a good method to ILZ from egg white. Hydrolysates of lysozyme were effective to inhibit lipid peroxidation in zebrafish model. Keywords: Lysozyme, Cation exchange chromatography, Enzymatic hydrolysis, Muramidase activity.