Study of Prepared α-Chymotrypsin as Enzyme Nanoparticles and of Biocatalytic Membrane Reactor

Abstract

Biocatalytic kinetic effect of α-chymotrypsin enzyme has been investigated in its free and pretreated forms (it was covered by a very thin, porous polymer layer, called enzyme nanoparticle) as well as its immobilized form into pores of polysulfone/polyamide asymmetric, hydrophilic membrane. Trimethoxysilyl and acrylamide-bisacrylamide polymers have been used for synthesis of enzyme nanoparticles. Applying Michaelis-Menten kinetics, the KM and vmax values of enzyme-polyacrylamide nanoparticles are about the same, as that of free enzyme. On the other hand, enzyme nanoparticles retain their activity 20–80 fold longer time period than that of the free enzyme, but their initial activity values are reduced to 13–55% of those of free enzymes, at 37 °C. Enzyme immobilized into asymmetric porous membrane layer remained active about 2.3-fold longer time period than that of native enzyme (at pH = 7.4 and at 23 °C), while its reaction rate was about 8-fold higher than that of free enzyme, measured in mixed tank reactor. The conversion degree of substrate was gradually decreased in presence of increasing convective flux of the inlet fluid phase. Biocatalytic membrane reactor has transformed 2.5 times more amount of substrate than the same amount of enzyme nanoparticles and 19 times more amount of substrate than free enzyme, measured in mixed tank reactor.