Synthesis and Characterization of Cross-Linked Aggregates of Peroxidase from <i>Megathyrsus maximus</i> (Guinea Grass) and Their Application for Indigo Carmine Decolorization
Angie V. Perez,
Jorge A. Gaitan-Oyola,
Diana P. Vargas-Delgadillo,
John J. Castillo,
Oveimar Barbosa,
Roberto Fernandez-Lafuente
Affiliations
Angie V. Perez
Grupo de Investigación en Materiales Porosos con Aplicaciones Ambientales y Tecnológicas, Departamento de Química, Universidad del Tolima, Ibagué 730006299, Colombia
Jorge A. Gaitan-Oyola
Grupo de Investigación en Materiales Porosos con Aplicaciones Ambientales y Tecnológicas, Departamento de Química, Universidad del Tolima, Ibagué 730006299, Colombia
Diana P. Vargas-Delgadillo
Grupo de Investigación en Materiales Porosos con Aplicaciones Ambientales y Tecnológicas, Departamento de Química, Universidad del Tolima, Ibagué 730006299, Colombia
John J. Castillo
Grupo de Investigación en Bioquímica y Microbiología, Escuela de Química, Universidad Industrial de Santander, Bucaramanga 680002, Colombia
Oveimar Barbosa
Grupo de Investigación en Materiales Porosos con Aplicaciones Ambientales y Tecnológicas, Departamento de Química, Universidad del Tolima, Ibagué 730006299, Colombia
Roberto Fernandez-Lafuente
Departamento de Biocatálisis, ICP-CSIC, Campus Cantoblanco UAM-CSIC, C/Marie Curie 2, 28049 Madrid, Spain
We present the synthesis of a cross-linking enzyme aggregate (CLEAS) of a peroxidase from Megathyrsus maximus (Guinea Grass) (GGP). The biocatalyst was produced using 50%v/v ethanol and 0.88%w/v glutaraldehyde for 1 h under stirring. The immobilization yield was 93.74% and the specific activity was 36.75 U mg−1. The biocatalyst surpassed by 61% the free enzyme activity at the optimal pH value (pH 6 for both preparations), becoming this increase in activity almost 10-fold at pH 9. GGP-CLEAS exhibited a higher thermal stability (2–4 folds) and was more stable towards hydrogen peroxide than the free enzyme (2–3 folds). GGP-CLEAS removes over 80% of 0.05 mM indigo carmine at pH 5, in the presence of 0.55 mM H2O2 after 60 min of reaction, a much higher value than when using the free enzyme. The operational stability showed a decrease of enzyme activity (over 60% in 4 cycles), very likely related to suicide inhibition.