Gonzalez F J, Korzekwa K R
Laboratory of Molecular Carcinogenesis, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
Annu Rev Pharmacol Toxicol. 1995;35:369-90. doi: 10.1146/annurev.pa.35.040195.002101.
Catalytically active cytochrome P450 enzymes have been successfully expressed in bacterial, yeast, and mammalian cells. A variety of expression vectors have been used, resulting in both transient and stable expression. The system of choice depends on the goals of a particular project. Factors such as expense, ease of use, and yields required should govern the decision whether to use bacterial, yeast, insect, or mammalian cDNA expression. High-level expression of mammalian P450s in bacteria usually requires modifications of the amino-terminal region of the enzyme. The Escherichia coli P450-OR fusion proteins may also come of age for use in fermentation-production processes for the chemical industry. Many cytochromes P450 have been expressed in yeast, with variable levels of expression. Baculovirus, albeit somewhat tedious in having to individualize expression conditions, can produce high levels of enzyme. The standard mammalian cell expression systems, both transient and stable, have been of tremendous value to drug metabolism and carcinogenesis research and will continue to play a role in these areas.
具有催化活性的细胞色素P450酶已在细菌、酵母和哺乳动物细胞中成功表达。人们使用了多种表达载体，实现了瞬时表达和稳定表达。选择何种系统取决于特定项目的目标。诸如费用、易用性和所需产量等因素应决定使用细菌、酵母、昆虫或哺乳动物cDNA表达的决策。哺乳动物P450s在细菌中的高水平表达通常需要对酶的氨基末端区域进行修饰。大肠杆菌P450-OR融合蛋白在化学工业的发酵生产过程中也可能会成熟起来。许多细胞色素P450已在酵母中表达，表达水平各不相同。杆状病毒尽管在确定表达条件方面有些繁琐，但能产生高水平的酶。标准的哺乳动物细胞表达系统，无论是瞬时的还是稳定的，对药物代谢和致癌作用研究都具有巨大价值，并将继续在这些领域发挥作用。
