Cell-based high-throughput screening of polysaccharide biosynthesis hosts

Microbial Cell Factories

Xue-Ping Guo¹  Si-Ling Huang¹  Zi-Xu Liu²  Feng-Shan Wang³  Ju-Zheng Sheng³  Jin Hou⁴ 

[1] Bloomage BioTechnology Corp., Ltd., 250010, Jinan, China;Key Laboratory of Chemical Biology of Natural Products (Ministry of Education), Institute of Biochemical and Biotechnological Drug, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, 250012, Jinan, China;Key Laboratory of Chemical Biology of Natural Products (Ministry of Education), Institute of Biochemical and Biotechnological Drug, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, 250012, Jinan, China;National Glycoengineering Research Center, Shandong University, 250012, Jinan, China;The State Key Laboratory of Microbial Technology, Shandong University, 250100, Jinan, China;

关键词: Polysaccharide;    Glycosyltransferase;    High-throughput screening;    Fluorescently-labeled substrate;    Biosensor;   

DOI  :  10.1186/s12934-021-01555-w

来源: Springer

【 摘 要 】

Valuable polysaccharides are usually produced using wild-type or metabolically-engineered host microbial strains through fermentation. These hosts act as cell factories that convert carbohydrates, such as monosaccharides or starch, into bioactive polysaccharides. It is desirable to develop effective in vivo high-throughput approaches to screen cells that display high-level synthesis of the desired polysaccharides. Uses of single or dual fluorophore labeling, fluorescence quenching, or biosensors are effective strategies for cell sorting of a library that can be applied during the domestication of industrial engineered strains and metabolic pathway optimization of polysaccharide synthesis in engineered cells. Meanwhile, high-throughput screening strategies using each individual whole cell as a sorting section are playing growing roles in the discovery and directed evolution of enzymes involved in polysaccharide biosynthesis, such as glycosyltransferases. These enzymes and their mutants are in high demand as tool catalysts for synthesis of saccharides in vitro and in vivo. This review provides an introduction to the methodologies of using cell-based high-throughput screening for desired polysaccharide-biosynthesizing cells, followed by a brief discussion of potential applications of these approaches in glycoengineering.

【 授权许可】

CC BY