We have previously introduced the use of permeabilized fission yeast cells (enzyme bags) that recombinantly express full-length CYPs for drug metabolism studies. Such enzyme bags are cells with pores that function as enzymes in situ. They can easily be prepared without a need for ultracentrifugation and may be used in similar protocols as microsomes. In this study we report the preparation of enzyme bag cocktails that permit the testing of multiple CYPs in a single enzyme bag reaction. Moreover, we established a convenient testing scheme that permits a rapid screen of all human CYPs for activity towards any given candidate substrate. An important aspect of this approach is the reduction of individual CYP test assays. If a cocktail containing many CYPs tests negative, it follows that all CYPs included in that cocktail need not be tested individually, thus saving time and resources. The new protocol was validated using two probe substrates.

之前，我们已经介绍了将重组表达全长CYP的透化裂变酵母细胞（酶袋）用于药物代谢研究的方法。这种酶袋是具有毛孔的细胞，这些毛孔在原位起酶的作用。它们可以容易地制备而无需超速离心，并且可以与微粒体类似的方案使用。在这项研究中，我们报告了酶袋混合物的制备，该混合物可在一个酶袋反应中测试多种CYP。此外，我们建立了方便的测试方案，可以快速筛选所有人类CYP对任何给定候选底物的活性。这种方法的一个重要方面是减少单个CYP测试方法。如果包含许多CYP的鸡尾酒测试为阴性，则无需对该鸡尾酒中包含的所有CYP进行单独测试，从而节省了时间和资源。使用两个探针基质验证了新方案。