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Characteristics of poly(AAc5-co-HPMA3-cl-EGDMA15) hydrogel-immobilized lipase of Pseudomonas aeruginosa MTCC-4713

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Abstract

Four series of noble networks were synthesized with acrylic acid (AAc) copolymerized with varying amount of 2-hydroxy propyl methacrylate or dodecyl methacrylate (AAc/HPMA or AAc/DMA; 5:1 to 5:5, w/w) in the presence of ethylene glycol dimethacrylate (EGDMA; 1, 5, 10, 15, and 20%, w/w) as a crosslinker and ammonium per sulfate (APS) as an initiator. Each of the networks was used to immobilize a purified lipase from Pseudomonasaeruginosa MTCC-4713. The lipase was purified by successive salting out with (NH4)2SO4, dialysis, and DEAE anion exchange chromatography. Two of the matrices, E15a, i.e. [poly (AAc5-co-DMA1-cl-EGDMA15)] and I15c, i.e. [poly (AAc5-co-HPMA3-cl-EGDMA15)], that showed relatively higher binding efficiency for lipase were selected for further studies. I15c-hydrogel retained 58.3% of its initial activity after 10th cycle of repetitive hydrolysis of p-NPP, and I15c was thus catalytically more stable and efficient than the other matrix. The I15c-hydrogel-immobilized enzyme showed maximum activity at 65°C and pH 9.5. The hydrolytic activity of free and I15c-hydrogel-immobilized enzyme increased profoundly in the presence of 5 mM chloride salts of Hg2+, NH4+, Al3+, K+, and Fe3+. The immobilized lipase was preferentially active on medium chain length p-nitrophenyl acyl ester (C:8, p-nitrophenyl caprylate). © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 100: 4636–4644, 2006

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