Comparing the effect of immobilization methods on the activity of lipase biocatalysts in ester hydrolysis
The activity of various lipases was compared, in both free and immobilized forms, using the kinetics of the hydrolysis reaction of p-nitrophenyl butyrate, which was followed with in situ UV/Vis diode array spectrophotometry. Several enzymes were used to catalyze the reaction, namely Candida antarctica lipase B and Fusarium solani pisi cutinase wildtype and three single-mutation variants. The enzymes were tested in three different forms: free, immobilized as cross-linked aggregates and supported on zeolite NaY. A simple kinetic model was used to allow a quantitative comparison of the behavior of the different catalysts. It was concluded that although immobilization reduces the activity of the enzyme, the zeolite offers a much higher specific activity when compared to the cross-linked aggregates, thus supplying a heterogeneous catalyst with promising catalytic properties.
KeywordsLipases Zeolite NaY Kinetic modeling Diode array spectrophotometry Spectral deconvolution
The authors gratefully thank Novozymes A/S (Denmark) for the kind gifts of Novozym® 435 and Lipozyme® CALB L. L. Costa and V. Brissos acknowledge Ph.D. grants (SFRH/BD/19108/2004 and SFRH/BD/9019/2002) from Fundação para a Ciência e Tecnologia.
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