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 suppo rt materials (NaY zeolite and polyamide-Accurel PA6). These preparatio ns were used to catalyze the hydrolysis of tricaprylin in a nonconvent ional medium. Water content is a very important factor in relation to enzyme activity in organic media, and the effect of the water in the r eaction medium as well as the effect of the organic solvent at differe nt water concentrations on the hydrolytic activity of the immobilized cutinase were investigated. Factors that are also important for the ca talytic activity of the enzyme such as immobilization pH, ionic streng th, 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 kine tic parameters of the immobilized enzyme were determined with triglyce rides containing fatty acids ranging from C-4 up to C-14. A selectivit y to C-8 was observed with both supports. The thermal stability of the enzyme was also investigated and a particularly good stability was ob served with cutinase immobilized on the NaY zeolite with no loss of in itial activity after 45 days. (C) 1997 by Elsevier Science Inc.