Effect of immobilization on stability and kinetic properties of alpha-L-fucosidase from Turbo cornutus.
An amount of alpha-L-fucosidase from T. cornutus liver was copolymerized with glutaraldehyde using bovine serum albumin as the carrier protein. The properties of the native, the soluble enzyme polymer complex, and the insoluble enzyme polymer complex were studied and compared under various conditions of pH, temperature, substrate, and inhibitor concentration. Native alpha-L-fucosidase was heat labile and lost more than 85% of its activity when incubated at 55 degrees C for 5 min. In contrast, under equivalent incubation conditions, both the soluble and the insoluble enzyme polymer complexes exhibited enhanced resistance to thermal inactivation and after 5 min lost only 65 and 40% of their original activity, respectively. Polymerzation also resulted in the shift of pH optima towards the acidic range, a decrease in activation energy and a change in the apparent K(m) values towards the p-nitrophenyl-alpha-L-fucopyranoside substrate.
Department of Biochemistry and Biophysics, University of Rhode Island, Kingston, Rhode Island 02881.
M A
MA
Cohenford
J C
JC
Urbanowski
J A
JA
Dain