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The Performance of Microbial Lipase Immobilized onto Ion Exchange Resins and onto the Natural Zeolite Clinoptilolite

Laurence Weatherley, Akash Anand


The paper describes a study into the performance of a microbial lipase immobilized on to four different ion exchangers. Lipase from the yeast Candida rugosa was immobilized onto the anionic ion exchange resins Dowex MWA-1, Purolite A109, and Amberlite IR45. The performance of the immobilized lipase in each case was evaluated by measuring the immobilization efficiency and by following the reaction kinetics of hydrolysis of a tri-glyceride ester to free fatty acid and glycerol. The immobilization efficiency is a measure of the enzyme activity in the immobilized state relative to the free enzyme. The performance of the three synthetic anionic ion exchangers was compared with that of the naturally occurring zeolitic cationic exchanger – clinoptilolite, which was similarly evaluated as a lipase support. The potential improvement of immobilization using the technique of pre-cross-linkage of the lipase using a glutaraldehyde crosslinking agent was also studied. The final part of the study focused on the potential enhancement of reaction performance through the addition of pure form cationic and anionic ion exchange resin to the reaction mixture together with the immobilized enzyme as a means of in-situ removal of free fatty acid product. In all cases the specific enzyme activity for the immobilized lipase was significantly lower compared with the activity in free solution. The Purolite A109 displayed the highest value of immobilization efficiency but only by a small margin compared with the Dowex MWA-1, and the Amberlite IR45. The clinoptilolite showed immobilization efficiency 50% lower than that of the synthetic ion exchangers. The application of glutaraldehyde cross-linkage during the immobilization protocol in the cases of the Dowex MWA-1 and the Purolite A109 resulted in increases in total protein uptake of 14.5 and 4.1%, respectively. The significant increase in total protein uptake in the case of the Dowex MWA-1 was not reflected in any significant enhancement of reaction kinetics. The reaction kinetics exhibited by the lipase immobilized on the anionic Purolite A109 resin, when mixed with cationic ion exchange resin in the sodium form during the reaction showed substantial enhancement. This observation suggests removal of fatty acid product through ion exchange uptake of H+ ions.

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