Bian Guangkai, Ma Tian, Liu Tiangang
Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education and School of Pharmaceutical Sciences, Wuhan University, Wuhan, PR China.
Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education and School of Pharmaceutical Sciences, Wuhan University, Wuhan, PR China; Hubei Engineering Laboratory for Synthetic Microbiology, Wuhan Institute of Biotechnology, Wuhan, PR China.
Methods Enzymol. 2018;608:97-129. doi: 10.1016/bs.mie.2018.04.025. Epub 2018 Jul 6.
Terpenoids represent a highly diverse group of natural products with wide applications. Engineering approaches have been used to increase titers of many value-added terpenoids, such as farnesene, taxadiene, lycopene, and astaxanthin. In this chapter, we review the in vitro reconstitution-based targeted engineering of terpenoids, as well as approaches for the mining of terpene cyclases and for increasing the chemical diversity. Information gained from in vitro reconstitution extends our understanding of the mechanisms underlying terpenoid biosynthesis, the contributions of enzymes and cofactors, and key enzymes and rate-limiting steps for the development of an ideal biosynthetic production system. The in vitro reconstitution-based targeted engineering strategy provides a rational and accurate engineering approach for terpenoid overproduction with high efficiency. Furthermore, an efficient terpenoid overproduction platform can accelerate the entire process for the mining of terpene cyclases and the discovery of novel terpenoids and can substantially increase the chemical diversity of these kinds of terpenoids.
萜类化合物是一类高度多样化的天然产物,具有广泛的应用。工程方法已被用于提高许多增值萜类化合物的产量,如法尼烯、紫杉二烯、番茄红素和虾青素。在本章中,我们综述了基于体外重构的萜类化合物靶向工程,以及萜烯环化酶的挖掘方法和增加化学多样性的方法。从体外重构中获得的信息扩展了我们对萜类化合物生物合成机制、酶和辅因子的作用以及开发理想生物合成生产系统的关键酶和限速步骤的理解。基于体外重构的靶向工程策略为高效过量生产萜类化合物提供了一种合理且准确的工程方法。此外,一个高效的萜类化合物过量生产平台可以加速萜烯环化酶的挖掘和新型萜类化合物的发现的整个过程,并能大幅增加这类萜类化合物的化学多样性。