Efficient immobilization of lipase from Candida rugosa by entrapment into poly(N-isopropylacrylamide-co-itaconic acid) hydrogels under mild conditions
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Temperature and pH-sensitive hydrogels, based on N-isopropylacrylamide and itaconic acid, with varying comonomer ratios and crosslinking agent content, were prepared by free radical crosslinking copolymerization. The immobilization of lipase from Candida rugosa was carried out by post-loading entrapment method at different temperatures until equilibrium swelling was achieved. The effects of the hydrogel composition and the immobilization temperature on the hydrogel-binding capacity, immobilized lipase specific activity, as well as the enzyme leakage were studied. It was found that the NiPAAm/IA ratio, crosslinking agent concentration and the temperature at which the entrapment was performed significantly affected the hydrogel-binding capacity. The biocatalysts obtained by entrapment into hydrogel with the highest itaconic acid content at 5 °C exhibited both highest binding capacity and the highest specific activity, but appeared to be less suitable for repeated uses than those obtained at 25 °C.
KeywordsLipase from Candida rugosa Immobilization temperature Hydrogel N-isopropylacrylamide Itaconic acid
The authors acknowledge funding from the Ministry of Education and Science of the Republic of Serbia, Project No. III 46010, “Novel encapsulation and enzyme technologies for designing of new biocatalysts and biologically active compounds targeting enhancement of food quality, safety and competitiveness” as well as Project No. 172062 “Synthesis and characterization of novel functional polymers and polymeric nanomaterials”. The authors would also like to thank Dr. Mirjana Mihajlović from the Institute for Biological Research “Siniša Stanković”, University of Belgrade, for her help in the sample lyophilization.
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