Peters Frank T, Bureik Matthias, Maurer Hans H
Institute of Forensic Medicine, Friedrich Schiller University of Jena, Fürstengraben 23, D-07740 Jena, Germany.
Bioanalysis. 2009 Jul;1(4):821-30. doi: 10.4155/bio.09.53.
Cytochrome P450 mono-oxygenases (CYPs) are the major enzymes involved in the metabolism of drugs and poisons in humans. The variation of their activity - due to genetic polymorphisms or enzyme inhibition/induction - potentially increases the risk of side effects or toxicity. Studies on CYP-dependent metabolism are important in drug-development or toxicity studies. Reference standards of drug metabolites required for such studies, especially in the context of metabolites in safety testing (MIST), are often not commercially available and their classical chemical synthesis can be cumbersome. Recently, a biotechnological approach using human CYP isozymes heterologously expressed in fission yeast was developed for the synthesis of drug metabolites. Among other aspects, this approach has the distinct advantages that the reactions run under mild conditions and that only the final product must be isolated and characterized. This review overviews the first practical applications of this new approach and discusses the selection of substrates, metabolites and fission yeast strains as well as important aspects of incubation, product isolation and clean-up.
细胞色素P450单加氧酶（CYPs）是参与人体药物和毒物代谢的主要酶。由于基因多态性或酶抑制/诱导导致其活性变化，可能会增加副作用或毒性风险。关于CYP依赖性代谢的研究在药物开发或毒性研究中很重要。此类研究所需的药物代谢物参考标准品，尤其是在安全性测试中的代谢物（MIST），通常无法从商业渠道获得，其经典化学合成可能很繁琐。最近，开发了一种利用在裂殖酵母中异源表达的人CYP同工酶合成药物代谢物的生物技术方法。在其他方面，该方法具有明显优势，即反应在温和条件下进行，且只需分离和表征最终产物。本综述概述了这种新方法的首次实际应用，并讨论了底物、代谢物和裂殖酵母菌株的选择以及孵育、产物分离和纯化的重要方面。
