Separation of lactoperoxidase using cation exchange nano-cryogels
Lactoperoxidase(LP)has received widespread attention in the fields of food,medicine and chemical industry because of its important biological functions and broad-spectrum antibacterial activity.In view of the shortcomings of traditional LP separation and purification methods,such as poor selectivity,high cost and easy contamination of separated materials,2-hydroxyethyl methacrylate and poly(glycidyl methacrylate)nanogels(PGN)were used as composite substrates to prepare poly(2-hydroxyethyl methacrylate)composite cryogel embedded with PGN nanogels(pHEMA/PGN-based cryogel)by using the principle of low-temperature freezing polymerization.Then the pHEMA/PGN-based cryogel was grafted with the functional monomer 2-acrylamino-2-methyl-1-propanesulfonic acid to obtain pHEMA/PGN cation exchange nano-cryogel(pHEMA/PGN nano-cryogel)for the chromatography separation of LP from whey.The basic properties of pHEMA/PGN nano-cryogel were investigated,and the effect of buffer pH on chromatography performance was emphatically explored.The results showed that the obtained pHEMA/PGN nano-cryogel had large pore structure with uniform distribution,high permeability,excellent mass transfer performance,and strong adsorption capacity for lysozyme reaching 4.41 mg·ml-1.The pHEMA/PGN nano-cryogel was used to isolate LP in whey,the purity was about 96.0％,the specific enzyme activity was 27.03 U·mg-1,and enzyme activity recovery was 86.5％under the optimal conditions.The excellent separation performance indicated that the pHEMA/PGN nano-cryogel has good potential in the field of lactoperoxidase separation from whey.
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