Mohd Faheem Khan, Carina Hof, Patricie Niemcova, Cormac D Murphy
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引用次数: 0
Abstract
Some species of the genus Cunninghamella (C. elegans, C. echinulata and C. blaskesleeana) produce the same phase I and phase II metabolites when incubated with xenobiotics as mammals, and thus are considered microbial models of mammalian metabolism. This had made these fungi attractive for metabolism studies with drugs, pesticides and environmental pollutants. As a substantial proportion of pharmaceuticals and agrochemicals are fluorinated, their biotransformation has been studied in Cunninghamella fungi and C. elegans in particular. This article details the methods employed for cultivating the fungi in planktonic and biofilm cultures, and extraction and analysis of fluorinated metabolites. Furthermore, protocols for the heterologous expression of Cunninghamella cytochromes P450 (CYPs), which are the enzymes associated with phase I metabolism, are described.
Cunninghamella 属的一些物种（C. elegans、C. echinulata 和 C. blaskesleeana）在与异种生物一起培养时会产生与哺乳动物相同的 I 期和 II 期代谢物，因此被认为是哺乳动物代谢的微生物模型。这使得这些真菌对药物、杀虫剂和环境污染物的代谢研究具有吸引力。由于相当一部分药物和农用化学品是含氟的，因此研究人员特别在坎宁安真菌和秀丽隐杆线虫中对它们的生物转化进行了研究。本文详细介绍了在浮游培养物和生物膜培养物中培养真菌以及提取和分析含氟代谢物的方法。此外，文章还介绍了与第一阶段代谢相关的酶--坎宁安菌细胞色素 P450（CYPs）的异源表达方案。
期刊介绍：
The critically acclaimed laboratory standard for almost 50 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Each volume is eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 500 volumes the series contains much material still relevant today and is truly an essential publication for researchers in all fields of life sciences, including microbiology, biochemistry, cancer research and genetics-just to name a few. Five of the 2013 Nobel Laureates have edited or contributed to volumes of MIE.
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