A novel cross-linked enzyme aggregates (CLEAs) of papain and neutrase-production, partial characterization and application.
The neutrase (EC 3.4.24.4) and papain (EC 3.4.22.2) were together immobilized ascross-linked enzyme aggregates (N-P-CLEAs) and their properties were characterized. The influence of the precipitant, cross-linking ratio of glutaraldehyde and cross-linking time were investigated. Ethanol was selected as the more efficient precipitant compared with ammonium sulfate. The proper cross-linking ratio of enzyme and glutaraldehyde was 1:5 (v/v) and the optimized cross-linking time was 4h. N-P-CLEAs showed obvious improvement in thermal stability and pH stability than the free enzyme (P<0.05) and could hold relatively high activity retention in nonpolar and hydrophilic solvents and without activity loss at 4°C for more than six months. The cross-linking reaction had been appeared in N-P-CLEAs and more orderly microscopic surface morphology of N-P-CLEAs was observed. The molecular weight and thermal denaturation temperature of N-P-CLEAs were increased while the isoelectric point was decreased compared with those of the free enzymes. Application of N-P-CLEAs in bean proteins and zein showed a higher degree of hydrolysis, such as the hydrolysis degree of mung bean protein hydrolyzed by N-P-CLEAs was 12%, increased by approximately 4.5% compared to that of free enzyme. The results demonstrated that the N-P-CLEAs was suitable for application in food protein hydrolysis.
['Chen|Zhongqin|Z|', 'Wang|Yanwei|Y|', 'Liu|Wei|W|', 'Wang|Jingya|J|', 'Chen|Haixia|H|']
D011232:Chemical Precipitation D003432:Cross-Linking Reagents D004795:Enzyme Stability D004800:Enzymes, Immobilized D005976:Glutaral D006868:Hydrolysis D007700:Kinetics D008666:Metalloendopeptidases D010206:Papain D010940:Plant Proteins D066329:Protein Aggregates D059748:Proteolysis D012997:Solvents
Cross-linked enzyme aggregates (CLEAs) Papain Characterization Neutrase Protein hydrolysis