Universidade Federal do Paraná, Departamento de Farmácia, Av. Pref. Lothário Meissner, 632, CEP 80210-170 Curitiba, PR, Brazil.
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.
J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Jan 15;945-946:10-6. doi: 10.1016/j.jchromb.2013.11.042. Epub 2013 Nov 25.
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.
通过不同的方法制备了漆酶固定化酶反应器（tyr-IMERs），并将其在线用于 HPLC 系统中，然后对其进行了比较。该酶从双孢蘑菇中提取，固定在使用不同条件制备的环氧-硅上。通过原位和分批技术进行酶固定化。从蛋白质和活性保留、IMERs 活性、动力学和稳定性方面比较了不同的方法。还研究了固定化方法对酶活性的影响以及 IMERs 对标准抑制剂的行为。使用微波辅助合成的环氧-硅原位固定化提供了制备漆酶 IMERs 的最佳条件。成功地用已知和潜在的漆酶抑制剂对 tyr-IMERs 进行了测试，结果表明它们可用于筛选该酶的抑制剂。
