title: Tyrosinase immobilized enzyme reactor: development and evaluation.
abstract: Immobilized enzyme reactors of tyrosinase (tyr-IMERs) for use on-line in HPLC system were prepared by different procedures and then compared. The enzyme, obtained from Agaricus bisporus, was immobilized on epoxy-silica which was prepared using different conditions. Enzyme immobilization was conducted by both in situ and in batch techniques. The different procedures were compared in terms of protein and activity retention, IMERs activity, kinetics and stability. The influence of immobilization procedure on enzyme activity and the behavior of the IMERs against a standard inhibitor were also investigated. In situ immobilization on epoxy-silica, synthesized using microwave assistance, provided the best conditions to prepare tyrosinase IMERs. The tyr-IMERs were successfully tested with known and potential inhibitors of tyrosinase, and the results showed that they can be used for the screening of inhibitors of that enzyme.
authors: Universidade Federal do Paraná, Departamento de Farmácia, Av. Pref. Lothário Meissner, 632, CEP 80210-170 Curitiba, PR, Brazil.
authors: Karina Bora
authors: KB
authors: de Oliveira
authors: Universidade Federal do Paraná, Departamento de Farmácia, Av. Pref. Lothário Meissner, 632, CEP 80210-170 Curitiba, PR, Brazil.
authors: Keylla Lençone
authors: KL
authors: Mischiatti
authors: Universidade Tecnológica Federal do Paraná, Programa de Pós-Graduação em Ciência e Tecnologia Ambiental, Rua Deputado Heitor Alencar Furtado, 4900, CEP 81280-340 Curitiba, PR, Brazil.
authors: José Domingos
authors: JD
authors: Fontana
authors: Universidade Federal do Paraná, Departamento de Química, Centro Politécnico CP 19081, CEP 81531-990 Curitiba, PR, Brazil. Electronic address: bho@ufpr.br.
authors: Brás Heleno
authors: BH
authors: de Oliveira
mesh_terms: D000364:Agaricus / Q000201:enzymology*; D019149:Bioreactors; D002851:Chromatography, High Pressure Liquid; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D014442:Monophenol Monooxygenase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D011753:Pyrones / Q000737:chemistry / Q000494:pharmacology; D012822:Silicon Dioxide / Q000737:chemistry
chemical_list: D004791:Enzyme Inhibitors; D004800:Enzymes, Immobilized; D011753:Pyrones; C011890:kojic acid; D012822:Silicon Dioxide; D014442:Monophenol Monooxygenase
keywords: Agaricus bisporus; Enzyme immobilization; Immobilized enzyme reactor; Tyrosinase