IMMOBILIZATION OF A RECOMBINANT CUTINASE BY ENTRAPMENT AND BY COVALENT BINDING - KINETIC AND STABILITY STUDIES

Fusarium solani pisi recombinant cutinase, immobilized by entrapment i n calcium alginate and by covalent binding on porous silica, was used to catalyze the hydrolysis of tricaprylin. The influence of relevant p arameters on the catalytic activity such as pH, temperature, and the s ubstrate concentration were studied. Cutinase immobilized by entrapmen t presented a Michaelis-Menten kinetics for tricaprylin concentrations up to 200 mM. At higher concentrations of substrate, inhibition was o bserved. For covalent binding immobilization, diffusional limitations were observed at low substrate concentrations and substrate inhibition occurred for concentrations higher than 150 mM. The stability of immo bilized cutinase was also evaluated. The enzyme immobilized by entrapm ent showed a high stability, in contrast to the immobilization on poro us silica.