高通量技术在萜类合成酶筛选中的应用

Application of high-throughput technology in the screening of terpene synthases

摘要:

萜类化合物种类繁多，生物活性多样，在食品、药品与化妆品等行业中具有广泛的应用。萜类化合物多来源于植物，然而随着合成生物学的快速发展，相较于传统的天然植物提取与化学合成方法，利用工程微生物进行萜类化合物异源合成的方法显得更为经济与环保。萜类合成酶的催化活性及合成产物的结构特性是萜类化合物异源合成的关键。通过蛋白定向进化与理性设计可以有针对性地优化萜类合成酶的催化性能及产物专一性，但该方案需要一个特异的筛选方法来实现蛋白突变体库的高通量筛选。近年来，一系列高通量筛选方法的建立使得萜类合成酶的筛选变得更加灵敏与高效。本文对近期建立的萜类合成酶高通量筛选方法进行了综述，简要概述了各种筛选方法的基本原理与优缺点，并对高通量筛选技术在萜类合成酶改造中的应用做出了展望。

Abstract:

Terpenoids, a group of natural products possessing diverse chemical structures and biological activities, have been widely used in the production of food additives, medicines, and cosmetics. Terpenoids are mainly produced by plants. With the rapid development of synthetic biology, heterologous production of terpenoids by engineered microbial strains is more economical and environmentally friendly compared with the conventional methods of extraction from plants and chemical synthesis. The catalytic activities of terpene synthases and the structural specificity of their products are pivotal for the heterologous biosynthesis of terpenoids. Directed evolution and rational design can be adopted to optimize the catalytic properties and product profile of target terpene synthases. However, this requires a feasible high-throughput method to screen mutant libraries. In recent years, various high-throughput screening methods have been developed to boost the sensitivity and efficiency in screening terpene synthases. We review recently established high-throughput screening methods for terpene synthases and briefly outline the principles and advantages and disadvantages of each method. Eventually, we envision the future directions of the application of high-throughput screening in the engineering of terpene synthases.