A new heterofunctional amino-vinyl sulfone support to immobilize enzymes: Application to the stabilization of β-galactosidase from Aspergillus oryzae

摘要

The paper shows the preparation of a new heterofunctional agarose support: amino-vinylsulfone. This has been employed to immobilize the interesting enzyme beta-galactosidase from Aspergillus oryzae. The enzyme cannot be immobilized on just vinylsulfone activated support a pH values ranging from 5.0 to 9.0. Neither the enzyme was immobilized using 200 mM of NaCl on amino-vinylsulfone support. However, the enzyme was readily immobilized at moderate ion strength at pH values from 5.0 to 9.0 via ion exchange on amino-vinylsulfone support, and later some covalent enzyme-support bonds could be formed, more rapidly at alkaline pH value. After optimization of immobilization pH, incubation pH and time, and blocking reagent, several immobilized biocatalysts on amino-vinylsulfone support having 50-80% of the initial activity and a stabilization factor of around 8-15 were prepared, depending on the exact immobilization conditions.