Key Laboratory for Potato Biology of Yunnan Province, The CAAS-YNNU-YINMORE Joint Academy of Potato Science, Yunnan Normal University, Kunming, China.
Lingnan Guangdong Laboratory of Modern Agriculture, Genome Analysis Laboratory of the Ministry of Agriculture, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen, China.
Plant Commun. 2019 Dec 3;1(1):100012. doi: 10.1016/j.xplc.2019.100012. eCollection 2020 Jan 13.
Cytochrome P450s (P450s) are the most versatile catalysts and are widely used by plants to synthesize a vast array of structurally diverse specialized metabolites that not only play essential ecological roles but also constitute a valuable resource for the development of new drugs. To accelerate the metabolic engineering of these high-value metabolites, it is imperative to identify and characterize pathway P450s, and to further improve their activities through protein engineering. In this review, we focus on P450 engineering and summarize the major strategies for enhancing the stability and activity of P450s and successful cases of P450 engineering. Studies in which the functions of P450s were altered to create metabolic pathways or novel compounds are discussed as well. We also overview emerging tools, specifically DNA synthesis, machine learning, and protein design, as well as the evolutionary patterns of P450s unveiled from a massive number of DNA sequences that could be integrated to accelerate the engineering of these enzymes. These approaches would greatly aid in the exploitation of plant-specialized metabolites or derivatives for various uses including medical applications.
细胞色素 P450s（P450s）是用途最广泛的催化剂，被广泛应用于植物合成结构多样的特殊代谢物，这些代谢物不仅发挥着重要的生态作用，而且是开发新药的宝贵资源。为了加速这些高价值代谢物的代谢工程，必须鉴定和表征途径 P450s，并通过蛋白质工程进一步提高它们的活性。在这篇综述中，我们重点关注 P450 的工程，并总结了提高 P450 稳定性和活性的主要策略以及 P450 工程的成功案例。我们还讨论了改变 P450 功能以创建代谢途径或新型化合物的研究。我们还概述了新兴工具，特别是 DNA 合成、机器学习和蛋白质设计，以及从大量 DNA 序列中揭示的 P450 的进化模式，这些工具可以整合在一起，加速这些酶的工程。这些方法将极大地有助于开发植物特殊代谢物或衍生物，用于包括医疗应用在内的各种用途。
