Behrendorff James B Y H, Huang Weiliang, Gillam Elizabeth M J
*Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St. Lucia, Brisbane, QLD 4072, Australia.
†School of Chemistry and Molecular Biosciences, The University of Queensland, St. Lucia, Brisbane, QLD 4072, Australia.
Biochem J. 2015 Apr 1;467(1):1-15. doi: 10.1042/BJ20141493.
Cytochrome P450 enzymes are renowned for their ability to insert oxygen into an enormous variety of compounds with a high degree of chemo- and regio-selectivity under mild conditions. This property has been exploited in Nature for an enormous variety of physiological functions, and representatives of this ancient enzyme family have been identified in all kingdoms of life. The catalytic versatility of P450s makes them well suited for repurposing for the synthesis of fine chemicals such as drugs. Although these enzymes have not evolved in Nature to perform the reactions required for modern chemical industries, many P450s show relaxed substrate specificity and exhibit some degree of activity towards non-natural substrates of relevance to applications such as drug development. Directed evolution and other protein engineering methods can be used to improve upon this low level of activity and convert these promiscuous generalist enzymes into specialists capable of mediating reactions of interest with exquisite regio- and stereo-selectivity. Although there are some notable successes in exploiting P450s from natural sources in metabolic engineering, and P450s have been proven repeatedly to be excellent material for engineering, there are few examples to date of practical application of engineered P450s. The purpose of the present review is to illustrate the progress that has been made in altering properties of P450s such as substrate range, cofactor preference and stability, and outline some of the remaining challenges that must be overcome for industrial application of these powerful biocatalysts.
细胞色素P450酶以其在温和条件下能够高度化学选择性和区域选择性地将氧插入多种化合物中的能力而闻名。这一特性在自然界中被用于多种生理功能，并且在所有生命王国中都已鉴定出这个古老酶家族的代表。P450酶的催化多功能性使其非常适合用于重新设计以合成精细化学品，如药物。尽管这些酶在自然界中并未进化以执行现代化学工业所需的反应，但许多P450酶显示出宽松的底物特异性，并对与药物开发等应用相关的非天然底物表现出一定程度的活性。定向进化和其他蛋白质工程方法可用于改善这种低水平的活性，并将这些混杂的通用酶转化为能够以精确的区域和立体选择性介导感兴趣反应的专一性酶。尽管在代谢工程中利用天然来源的P450酶取得了一些显著成功，并且P450酶已被反复证明是工程设计的优良材料，但迄今为止，工程化P450酶的实际应用实例很少。本综述的目的是阐述在改变P450酶的性质（如底物范围、辅因子偏好和稳定性）方面所取得的进展，并概述这些强大的生物催化剂在工业应用中必须克服的一些剩余挑战。
