RT Journal Article
T1 Changes in ficin specificity by different substrate proteins promoted by enzyme immobilization
A1 González Vásquez, Alex D.
A1 Hocine, El Siar
A1 Urzúa, Marcela
A1 Rocha Martín, Javier
A1 Fernández Lafuente, Roberto
AB 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.
PB Elsevier
SN 0141-0229
YR 2024
FD 2024
LK https://hdl.handle.net/20.500.14352/112066
UL https://hdl.handle.net/20.500.14352/112066
LA eng
NO Gonzalez-Vasquez, A. D., Hocine, E. S., Urzúa, M., Rocha-Martin, J., & Fernandez-Lafuente, R. (2024). Changes in ficin specificity by different substrate proteins promoted by enzyme immobilization. Enzyme and Microbial Technology, 181. https://doi.org/10.1016/J.ENZMICTEC.2024.110517
NO We gratefully recognize the financial support from Ministerio de Ciencia e Innovación and Agencia Estatal de Investigación (Spanish Government) (PID2022–136535OB-I00)., FONDECYT Project N° 1231631(MU), Beca Doctorado Nacional N° 21212403 Agencia Nacional de Investigación y Desarrollo (ANID). Siar thanks Algerian Ministry of higher education and scientific research. Also, thank INATAA, University of Constantine 1, Brothers Mentouri, Algeria.
NO Ministerio de Ciencia e Innovación (España)
NO Agencia Nacional de Investigación y Desarrollo (Chile)
NO Algerian Ministry of Higher Education and Scientific Research
NO Institute of Nutrition and Agro-Food Technologies (Argelia)
NO Fondo Nacional de Desarrollo Científico y Tecnológico (Chile)
DS Docta Complutense
RD 7 abr 2025