title: Construction of polyketide overproducing Escherichia coli strains via synthetic antisense RNAs based on in silico fluxome analysis and comparative transcriptome analysis.
abstract: Rapid assessment and optimization of the incompatible metabolic modules remain a challenge. Here, we developed a systematic approach to characterize the module interactions and improve the problematic modules during the 6-deoxyerythronolide B (6dEB) biosynthesis in E. coli. Tremendous differences in the overall trends of flux changes of various metabolic modules were firstly uncovered based on in silico fluxome analysis and comparative transcriptome analysis. Potential targets for improving 6dEB biosynthesis were identified through analyzing these discrepancies. All 25 predicted targets at modules of PP pathway and nucleotide metabolism were firstly tested for improving the 6dEB production in E. coli via synthetic antisense RNAs. Down-regulation of 18 targets genes leads to more than 20% increase in 6dEB yield. Combinatorial repression of targets with greater than 60% increase in 6dEB titer, e.g., anti-guaB/anti-zwf led to a 296.2% increase in 6dEB production (210.4 mg/L in flask) compared to the control (53.1 mg/L). This is the highest yield yet reported for polyketide heterologous biosynthesis in E. coli. This study demonstrates a strategy to enhance the yield of heterologous products in the chassis cell and indicates the effectiveness of antisense RNA for use in metabolic engineering.
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.|Bioengineering Research Center, Guangzhou Institute of Advanced Technology, Chinese Academy of Sciences, Guangzhou, China.
authors.forename: Hai-Lin
authors.initials: HL
authors.lastname: Meng
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
authors.forename: Zhi-Qiang
authors.initials: ZQ
authors.lastname: Xiong
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.|Food Science and Engineering, College of Food Science and Technology, Shanghai Ocean University, Shanghai, China.
authors.forename: Shu-Jie
authors.initials: SJ
authors.lastname: Song
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Wang
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China. yongwang@sibs.ac.cn.
authors.forename: Yong
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000900:Anti-Bacterial Agents / Q000096:biosynthesis; D003198:Computer Simulation; D015536:Down-Regulation; D004917:Erythromycin / Q000031:analogs & derivatives* / Q000096:biosynthesis; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods*; D015964:Gene Expression Regulation, Bacterial; D060847:Metabolic Engineering / Q000379:methods*; D010427:Pentose Phosphate Pathway / Q000187:drug effects; D016372:RNA, Antisense / Q000494:pharmacology*
chemical_list: D000900:Anti-Bacterial Agents; D029968:Escherichia coli Proteins; D016372:RNA, Antisense; C034277:6-deoxyerythronolide B; D004917:Erythromycin
keywords: Antisense RNA; Comparative transcriptome analysis; In silico fluxome analysis; Metabolic module interactions; Polyketides