Changes in ficin specificity by different substrate proteins promoted by enzyme immobilization.

Autor: Gonzalez-Vasquez AD; Departamento de Biocatalisis, ICP-CSIC, Campus UAM-CSIC, Madrid 28049, Spain; Departamento de Química, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Casilla 653, Santiago, Ñuñoa 7800003, Chile., Hocine ES; Departamento de Biocatalisis, ICP-CSIC, Campus UAM-CSIC, Madrid 28049, Spain; Agri-food Engineering Laboratory (GENIAAL), Institute of Food, Nutrition and Agri-Food Technologies (INATAA), University of Brothers Mentouri Constantine 1, Algeria., Urzúa M; Departamento de Química, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Casilla 653, Santiago, Ñuñoa 7800003, Chile., Rocha-Martin J; Department of Biochemistry and Molecular Biology, Faculty of Biology, Complutense University of Madrid, José Antonio Novais 12, Madrid 28040, Spain. Electronic address: javrocha@ucm.es., Fernandez-Lafuente R; Departamento de Biocatalisis, ICP-CSIC, Campus UAM-CSIC, Madrid 28049, Spain. Electronic address: rfl@icp.csic.es.
Jazyk: angličtina
Zdroj: Enzyme and microbial technology [Enzyme Microb Technol] 2024 Dec; Vol. 181, pp. 110517. Date of Electronic Publication: 2024 Sep 21.
DOI: 10.1016/j.enzmictec.2024.110517
Abstrakt: Ficin extract has been immobilized using different supports: glyoxyl and Aspartic/1,6 hexamethylenediamine (Asp/HA) agarose beads. The latter was later submitted to glutaraldehyde modification to get covalent immobilization. The activities of these 3 kinds of biocatalysts were compared utilizing 4 different substrates, casein, hemoglobin and bovine serum albumin and benzoyl-arginine-p-nitroanilide at pH 7 and 5. Using glyoxyl-agarose, the effect of enzyme-support reaction time on the activity versus the four substrates at both pH values was studied. Reaction time has been shown to distort the enzyme due to an increase in the number of covalent support-enzyme bonds. Surprisingly, for all the substrates and conditions the prolongation of the enzyme-support reaction did not imply a decrease in enzyme activity. Using the Asp/HA supports (with different amount of HA) differences in the effect on enzyme activity versus the different substrates are much more significant, while with some substrates the immobilization produced a decrease in enzyme activity, with in other cases the activity increased. These different effects are even increased after glutaraldehyde treatment. That way, the conformational changes induced by the biocatalyst immobilization or the chemical modification fully altered the enzyme protein specificity. This may also have some implications when following enzyme inactivation.
(Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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