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RevistaPROCESS BIOCHEMISTRY
Año2012
Volumen47
Páginas377-2382
Internacional

CHEMICAL AMINATION OF LIPASE B FROM Candida antarctica IS AN EFFICIENT SOLUTION FOR THE PREPARATION OF CROSSLINKED ENZYME AGGREGATES

Autores:Roberto Fernandez-Lafuente
Grupos de investigación:Optimización de biocatalizadores y bioprocesos enzimáticos
Magaly Galvis1, Oveimar Barbosa1,  Monica Ruiz1, Jennifer Cruz1, Claudia Ortiz2,
Rodrigo Torres1,* and Roberto Fernandez-Lafuente3,*
1Escuela de Química, Grupo de Investigación en Bioquímica y Microbiología (GIBIM), Universidad Industrial de Santander, Bucaramanga, Colombia
2 Escuela de Bacteriología y Laboratorio Clínico, Universidad Industrial de Santander, Bucaramanga, Colombia
Departamento de Biocatálisis, Instituto de Catálisis-CSIC, Campus UAM-CSIC, C/Marie Curie 2, Madrid, Cantoblanco 28049, Spain

Lipase B from Candida antarctica (CALB) is not very adequate to prepare cross-linked enzyme aggregates (CLEAS). Although the precipitation step is easy using different precipitants, the cross-linking step becomes a problem due to the low amount of Lys residues in this enzyme. In this paper, we have enriched the enzyme in amino groups by chemical amination of the enzyme using ethylenediamine and carbodiimide. The modification was performed using a solid phase strategy modifying the enzyme adsorbed on octyl-Sepharose. After desorption from the support, the enzyme was more active at pH 7.0 than the unmodified enzyme. This modified enzyme showed to be suitable to produce CLEAs. Using this modified enzyme, precipitation is also effective but the crosslinking step did not fail in giving an intense intermolecular crosslinking. This way, the CLEA did not release enzyme molecules even if boiled in SDS. Stability of this CLEA was higher in both thermal and cosolvent inactivation experiments than that of the coCLEA produced by coagregation of BSA and CALB; another alternative to produce a CLEA of this interesting enzyme.
The strategy may be of high interest for many other enzymes as a way to both permit the production of CLEAs and to improve enzyme stability during CLEA production.
Palabras clave:CALB, CLEA, chemical amination, enzyme crosslinking
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