title: Chemical modification of turnip peroxidase with methoxypolyethylene glycol enhances activity and stability for phenol removal using the immobilized enzyme.
abstract: Peroxidase from turnip roots (TP) was isolated followed by modification with methoxypolyethylene glycol (MPEG). The catalytic activity of the modified TP (MTP) on ABTS increased 2.5 times after 80 min of reaction. MTP showed a KM similar value to that of TP, but a significantly greater kcat for ABTS oxidation, in aqueous buffer. Chemical modification produced an enhanced stability in organic solvents and increased thermal stability of about 4 times that of TP, in aqueous buffer at 70 degrees C. Circular dichroism showed that MPEG modification decreased TP alpha-helical structure from 26 to 16% and increased beta-turns from 26 to 34%, resulting in an enhanced conformational stability. The temperature at the midpoint of thermal denaturation (melting temperature) increased from 57 to 63 degrees C after modification. MTP was immobilized in alginate beads (IMTP) and tested for oxidative polymerization of concentrated phenolic synthetic solutions, achieving 17 effective contact cycles removing >65% phenols. IMTP may be useful for the development of an enzymatic process for wastewater effluent treatment.
authors.affiliation: Departamento de Investigación y Posgrado en Alimentos PROPAC, Facultad de Química, Universidad Autónoma de Querétaro, C.U. Cerro de las Campanas s/n, Querétaro, Qro. 76010, Mexico.
authors.forename: F
authors.initials: F
authors.lastname: Quintanilla-Guerrero
authors.forename: M A
authors.initials: MA
authors.lastname: Duarte-Vázquez
authors.forename: R
authors.initials: R
authors.lastname: Tinoco
authors.forename: M
authors.initials: M
authors.lastname: Gómez-Suárez
authors.forename: B E
authors.initials: BE
authors.lastname: García-Almendárez
authors.forename: R
authors.initials: R
authors.lastname: Vazquez-Duhalt
authors.forename: C
authors.initials: C
authors.lastname: Regalado
mesh_terms: D029688:Brassica napus / Q000201:enzymology*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized; D006358:Hot Temperature; D007700:Kinetics; D009195:Peroxidase / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism*; D010636:Phenols / Q000378:metabolism*; D018517:Plant Roots / Q000201:enzymology*; D011092:Polyethylene Glycols / Q000494:pharmacology*; D011487:Protein Conformation
chemical_list: D004800:Enzymes, Immobilized; D010636:Phenols; D011092:Polyethylene Glycols; C028210:monomethoxypolyethylene glycol; D009195:Peroxidase