Development of a dual temperature control system for isoprene biosynthesis in Saccharomyces cerevisiae  

作　　者：Jiaxi Lin Zhen Yao Xiaomei Lyu Lidan Ye Hongwei Yu 

机构地区：[1]Institute of Bioengineering,College of Chemical and Biological Engineering,Zhejiang University,Hangzhou 310027,China [2]Hangzhou Global Scientific and Technological Innovation Center,Zhejiang University,Hangzhou 311200,China [3]Key Laboratory of Systems Microbial Biotechnology,Tianjin Institute of Industrial Biotechnology,Chinese Academy of Sciences,Tianjin 300308,China [4]School of Food Science and Technology,Jiangnan University,Wuxi 214122,China

出　　处：《Frontiers of Chemical Science and Engineering》2022年第7期1079-1089,共11页化学科学与工程前沿（英文版）

基　　金：financially supported by the National Key Research and Development Program of China(Grant Nos.2018YFA0901800 and 2020YFA0908400);the National Natural Science Foundation of China(Grant No.21776244);Zhejiang Provincial Natural Science Foundation of China(Grant No.LZ20B060002).

摘　　要：Conflict between cell growth and product accumulation is frequently encountered in the biosynthesis of secondary metabolites. To address the growth-production conflict in yeast strains harboring the isoprene synthetic pathway in the mitochondria, the dynamic control of isoprene biosynthesis was explored. A dual temperature regulation system was developed through engineering and expression regulation of the transcriptional activator Gal4p. A cold-sensitive mutant, Gal4ep19, was created by directed evolution of Gal4p based on an internally developed growth-based high-throughput screening method and expressed under the heat-shock promoter PSSA4 to control the expression of PGAL-driven pathway genes in the mitochondria. Compared to the control strain with constitutively expressed wild-type Gal4p, the dual temperature regulation strategy led to 34.5% and 72% improvements in cell growth and isoprene production, respectively. This study reports the creation of the first cold-sensitive variants of Gal4p by directed evolution and provides a dual temperature control system for yeast engineering that may also be conducive to the biosynthesis of other high-value natural products.

关 键 词：transcriptional activator directed evolution dynamic control HEAT-SHOCK ISOPRENE 

分 类 号：O63[理学—高分子化学]