Reutilization of the Most Stable Coimmobilized Enzyme Using Glutaraldehyde Chemistry to Produce a New Combi-biocatalyst When the Coimmobilized Enzyme with a Lower Stability Is Inactivated

Abstract

In the present article, glutaraldehyde was used to covalently coimmobilize the lipase Eversa Transform 2.0 and the β-galactosidase from Aspergillus oryzae. Both enzymes were adsorbed on amino supports and modified with glutaraldehyde. However, the first enzyme remained almost fully active under stress conditions, while the β-galactosidase lost a large percentage of its activity. To prevent the necessity of discarding both enzymes, the lipase was covalently immobilized following this immobilization strategy. The biocatalyst was reduced to eliminate its chemical reactivity, and the β-galactosidase was then coimmobilized via ion exchange. The incubation at high concentrations of salt desorbed the β-galactosidase from the support. This combi-biocatalyst was used in three inactivation/rebuilding cycles where the inactivated β-galactosidase was liberated from the combi-biocatalyst by washing at high ionic strength and replaced with a fresh enzyme, while the immobilized lipase maintained its activity throughout the 3 cycles. That way, it was possible to use this strategy to reuse the immobilized Eversa Transform 2.0 to build new combi-biocatalysts after β-galactosidase inactivation.

A Preblocking Strategy with Rapid Immobilization of Nitrile Hydratase and Precise Control of Hydrophobic Microenvironment for High Regioselective Amide Transformation

“…Enzyme immobilization could be combined with related strategies (gene editing, chemical modification, separation techniques) to cater for different applications properly. − Notably, the control of mass transfer and partitioning expands the possibilities for biocatalysis of hydrophobic substrates . Consequently, there’s a pressing need for more specific enzyme immobilization platforms …”