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RevistaPROCESS BIOCHEMISTRY
Año2016
Volumen51
Páginas1391-1396
Internacional

REUSE OF ANION EXCHANGERS AS SUPPORTS FOR ENZYME IMMOBILIZATION: REINFORCEMENT OF THE ENZYME-SUPPORT MULTIINTERACTION AFTER ENZYME INACTIVATION

Autores:V.C: Corberan, Roberto Fernandez-Lafuente
Grupos de investigación:Optimización de biocatalizadores y bioprocesos enzimáticos
Jose J. Virgen-Ortíz a+, Sara Peirce a,b+, Veymar G. Tacias-Pascacio a,c, Vicente Cortes-Corberan a, Antonio Marzocchella b, Maria Elena Russo d, Roberto Fernandez-Lafuente a*
 
a Departamento de Biocatálisis. Instituto de Catálisis-CSIC, Campus UAM-CSIC, Cantoblanco, 28049 Madrid, Spain.
b Dipartimento di Ingegneria Chimica, dei Materiali e della Produzione Industriale. Universita' degli Studi di Napoli Federico II, Italy.
c Unidad de Investigación y Desarrollo en Alimentos. Instituto Tecnológico de Veracruz, Calzada Miguel A. de Quevedo 2779, 91897 Veracruz, Mexico.
d Istituto di Ricerche sulla Combustione– Consiglio Nazionale delle Ricerche, Napoli, Italy.


β-galactosidase from Aspergillus oryze has been immobilized on agarose beads coated with polyethyleneimine. The fresh enzyme was released from the support using 500 mL NaCl at pH  7. After thermal inactivation or inactivation in the presence of organic solvents, the active enzyme still could be easily released from the support using similar conditions. However, SDS-PAGE analysis of the enzyme contained in the support after enzyme desorption showed that enzyme molecules remained in the support (inactivated enzyme molecules). This effect was stronger on enzyme preparations inactivated in organic medium. Now the conditions should be greatly strengthen to permit the full enzyme desorption: only after incubation in 2 M sodium phosphate at pH 2 and 50ºC full release of the enzyme molecules was achieved. This could be repeated several cycles with any difference neither in the immobilization performance nor on the SDS-PAGE analysis. Therefore, the reversibility of the immobilization is a real fact, but recovery of a support fully free of protein molecules is not an easy objective after enzyme inactivation, because the inactivated enzymes seemed to unfold increasing in a great way the interaction with the support, driving to a very strong enzyme-support multi-interaction that difficulty its desorption.
Palabras clave:enzyme unfolding, ion exchange, reversibly immobilization, support-enzyme multi-interaction, enzyme desorption, support reuse
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