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

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 stre...

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Detalles Bibliográficos
Autores: Carballares, Diego, Abellanas-Pérez, Pedro, Andrades, Diandra de, Polizeli, Maria de Lourdes T. M., Rocha-Martín, Javier, Fernández-Lafuente, Roberto
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2024
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/390001
Acceso en línea:http://hdl.handle.net/10261/390001
https://api.elsevier.com/content/abstract/scopus_id/85191406786
Access Level:acceso abierto
Palabra clave:Dissimilar enzyme stabilities
Enzyme coimmobilization
Enzyme release from combi-biocatalysts | reuse of stable coimmobilized enzymes
Descripción
Sumario: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.