Effect of the immobilization support on the hydrolytic activity of a cutinase from Fusarium solani pisi

Fusarium solani pisi recombinant cutinase was immobilized on two support materials (NaY zeolite and polyamide-Accurel PA6). These preparations were used to catalyze the hydrolysis of tricaprylin in a nonconventional medium. Water content is a very important factor in relation to enzyme activity in organic media, and the effect of the water in the reaction medium as well as the effect of the organic solvent at different water concentrations on the hydrolytic activity of the immobilized cutinase were investigated. Factors that are also important for the catalytic activity of the enzyme such as immobilization pH, ionic strength, and pH of the reaction medium and temperature were optimized. The selected conditions for further work were pH 9 (the immobilization pH) and 30 degrees C. In order to evaluate cutinase selectivity, the kinetic parameters of the immobilized enzyme were determined with triglycerides containing fatty acids ranging from C-4 up to C-14. A selectivity to C-8 was observed with both supports. The thermal stability of the enzyme was also investigated and a particularly good stability was observed with cutinase immobilized on the NaY zeolite with no loss of initial activity after 45 days. (C) 1997 by Elsevier Science Inc.