Multi-strategy engineering unusual sugar TDP-L-mycarose biosynthesis to improve the production of 3-O-α-mycarosylerythronolide B in Escherichia coli  

作　　者：Zhifeng Liu Jianlin Xu Zhanguang Feng Yong Wang 

机构地区：[1]CAS-Key Laboratory of Synthetic Biology,CAS Centre for Excellence in Molecular Plant Sciences,Institute of Plant Physiology and Ecology,Chinese Academy of Sciences,Shanghai,200032,China [2]University of Chinese Academy of Sciences,Beijing,100039,China [3]State Key Laboratory of Bioreactor Engineering,East China University of Science and Technology,Shanghai,200237,China

出　　处：《Synthetic and Systems Biotechnology》2022年第2期756-764,共9页合成和系统生物技术（英文）

基　　金：This study was financially supported by the National Key R&D Program of China(2018YFA0900600);the Program of Shanghai Academic Research Leader(20XD1404400);the Strategic Priority Research Program“Molecular mechanism of Plant Growth and Development”of CAS(XDB27020202);the National Natural Science Foundation of China(31670099);the Construction of the Registry and Database of Bioparts for Synthetic Biology of the Chinese Academy of Science(No.ZSYS-016);the International Partnership Program of Chinese Academy of Science(No.153D31KYSB20170121);the National Key Laboratory of Plant Molecular Genetics,SIPPE,CAS.

摘　　要：The insufficient supply of sugar units is the key limitation for the biosynthesis of glycosylated products.The unusual sugar TDP-L-mycarose is initially attached to the C3 of the polyketide erythronolide B,resulting in 3-O-α-mycarosylerythronolide B(MEB).Here,we present the de novo biosynthesis of MEB in Escherichia coli and improve its production using multi-strategy metabolic engineering.Firstly,by blocking precursor glucose-1-phosphate competing pathways,the MEB titer of triple knockout strain QC13 was significantly enhanced to 41.2 mg/L,9.8-fold to that produced by parental strain BAP230.Subsequently,the MEB production was further increased to 48.3 mg/L through overexpression of rfbA and rfbB.Moreover,the CRISPRi was implemented to promote the TDP-L-mycarose biosynthesis via repressing the glycolysis and TDP-L-rhamnose pathway.Our study paves the way for efficient production of erythromycins in E.coli and provides a promising platform that can be applied for biosynthesis of other glycosylated products with unusual sugars.

关 键 词：TDP-L-mycarose Glycosylated products 3-O-α-mycarosylerythronolide B Metabolic engineering CRISPRi 

分 类 号：Q78[生物学—分子生物学]