Enzymatic and Antibacterial Activity of Low-pH and Pepsin-Modified Freeze-Dried Lysozyme from Local Chicken Egg White
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antibacterial, egg white, enzyme activity, lysozyme, MIC
Abstract
The research aimed to isolate and modify lysozyme from local chicken egg white to improve the antibacterial activity of lysozyme hydrolysates, and to characterize the isolated and modified lysozyme. Lysozyme was isolated from local chicken egg white using an ion-exchange resin. The lysozyme isolate was then modified with pepsin in a low-pH solution, and the pepsin-modified samples were neutralized to pH 7 before testing. The enzymatic activity of lysozyme was measured spectro-photometrically using Micrococcus lysodeikticus. The antibacterial activity of lysozyme was tested using the microdilution method. The lysozyme was characterized by SDS-PAGE and RP-HPLC. The yield of lysozyme isolate was 5.35±0.23% (n= 3) with a lysozyme concentration of 41.07±4.44 mg/g (n= 3). After dialysis, the lysozyme concentration was 143.24±8.32 mg/g (n= 3). The enzymatic activity of the lysozyme isolate was 4,778±1,835 unit/mg (n= 3). The modification of lysozyme improved its antibacterial activity by decreasing its MIC value from 6 mg/mL (or >6 mg/mL) to 3 mg/mL and increasing its antibacterial spectrum to include Bacillus cereus, Staphylococcus aureus, Salmonella Typhimurium, and Escherichia coli. However, all low-pH treatments against E. coli reduced the MIC value only from >6 mg/mL to 6 mg/mL. The SDS-PAGE and RP-HPLC profiles of the lysozyme isolate showed that it had the same molecular weight (MW) and retention time (RT) as the standard lysozyme, with values of approximately 14.70±0.43 kDa (n= 3) and 43.59±0.09 min (n= 3), respectively. The modification of lysozyme also affected its enzymatic activity and SDS-PAGE and RP-HPLC profiles, explaining why the antibacterial activity increased.
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