title: A novel cross-linked enzyme aggregates (CLEAs) of papain and neutrase-production, partial characterization and application.
abstract: 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.
authors.affiliation: Tianjin Key Laboratory for Modern Drug Delivery & High-Efficiency, School of Pharmaceutical Science and Technology, Tianjin University, Tianjin, 300072, PR China.
authors.forename: Zhongqin
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Tianjin Key Laboratory for Modern Drug Delivery & High-Efficiency, School of Pharmaceutical Science and Technology, Tianjin University, Tianjin, 300072, PR China.
authors.forename: Yanwei
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Tianjin Key Laboratory for Modern Drug Delivery & High-Efficiency, School of Pharmaceutical Science and Technology, Tianjin University, Tianjin, 300072, PR China.
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.affiliation: Tianjin Key Laboratory for Modern Drug Delivery & High-Efficiency, School of Pharmaceutical Science and Technology, Tianjin University, Tianjin, 300072, PR China.
authors.forename: Jingya
authors.initials: J
authors.lastname: Wang
authors.affiliation: Tianjin Key Laboratory for Modern Drug Delivery & High-Efficiency, School of Pharmaceutical Science and Technology, Tianjin University, Tianjin, 300072, PR China. Electronic address: chenhx@tju.edu.cn.
authors.forename: Haixia
authors.initials: H
authors.lastname: Chen
mesh_terms: D011232:Chemical Precipitation; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D005976:Glutaral / Q000494:pharmacology; D006868:Hydrolysis; D007700:Kinetics; D008666:Metalloendopeptidases / Q000737:chemistry* / Q000378:metabolism; D010206:Papain / Q000737:chemistry* / Q000378:metabolism; D010940:Plant Proteins / Q000378:metabolism; D066329:Protein Aggregates; D059748:Proteolysis; D012997:Solvents / Q000737:chemistry
chemical_list: D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized; D010940:Plant Proteins; D066329:Protein Aggregates; D012997:Solvents; D010206:Papain; D008666:Metalloendopeptidases; C044194:microbial metalloproteinases; D005976:Glutaral
keywords: Characterization; Cross-linked enzyme aggregates (CLEAs); Neutrase; Papain; Protein hydrolysis