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

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.