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

Abstract

AbstractFour 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 Pseudomonas aeruginosa 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