title: Spatial organization of heterologous metabolic system in vivo based on TALE.
abstract: For years, prokaryotic hosts have been widely applied in bio-engineering. However, the confined in vivo enzyme clustering of heterologous metabolic pathways in these organisms often results in low local concentrations of enzymes and substrates, leading to a low productive efficacy. We developed a new method to accelerate a heterologous metabolic system by integrating a transcription activator-like effector (TALE)-based scaffold system into an Escherichia coli chassis. The binding abilities of the TALEs to the artificial DNA scaffold were measured through ChIP-PCR. The effect of the system was determined through a split GFP study and validated through the heterologous production of indole-3-acetic acid (IAA) by incorporating TALE-fused IAA biosynthetic enzymes in E. coli. To the best of our knowledge, we are the first to use the TALE system as a scaffold for the spatial organization of bacterial metabolism. This technique might be used to establish multi-enzymatic reaction programs in a prokaryotic chassis for various applications.
authors.affiliation: College of Science, National University of Defense Technology, Changsha, Hunan, 410073, People's Republic of China.
authors.forename: Lv-yun
authors.initials: LY
authors.lastname: Zhu
authors.affiliation: College of Aerospace Science and Engineering, National University of Defense Technology, Changsha, Hunan, 410073, People's Republic of China.
authors.forename: Xin-Yuan
authors.initials: XY
authors.lastname: Qiu
authors.affiliation: College of Science, National University of Defense Technology, Changsha, Hunan, 410073, People's Republic of China.
authors.forename: Ling-Yun
authors.initials: LY
authors.lastname: Zhu
authors.affiliation: College of Science, National University of Defense Technology, Changsha, Hunan, 410073, People's Republic of China.
authors.forename: Xiao-Min
authors.initials: XM
authors.lastname: Wu
authors.affiliation: College of Aerospace Science and Engineering, National University of Defense Technology, Changsha, Hunan, 410073, People's Republic of China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: College of Aerospace Science and Engineering, National University of Defense Technology, Changsha, Hunan, 410073, People's Republic of China.
authors.forename: Qian-Hui
authors.initials: QH
authors.lastname: Zhu
authors.affiliation: College of Science, National University of Defense Technology, Changsha, Hunan, 410073, People's Republic of China.
authors.forename: Dong-Yu
authors.initials: DY
authors.lastname: Fan
authors.affiliation: College of Science, National University of Defense Technology, Changsha, Hunan, 410073, People's Republic of China.
authors.forename: Chu-Shu
authors.initials: CS
authors.lastname: Zhu
authors.affiliation: College of Science, National University of Defense Technology, Changsha, Hunan, 410073, People's Republic of China.
authors.forename: Dong-Yi
authors.initials: DY
authors.lastname: Zhang
mesh_terms: D053898:Biosynthetic Pathways / Q000235:genetics*; D004247:DNA / Q000378:metabolism; D004798:Enzymes / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism; D007210:Indoleacetic Acids / Q000378:metabolism*; D060847:Metabolic Engineering; D009097:Multienzyme Complexes / Q000235:genetics* / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D004798:Enzymes; D029968:Escherichia coli Proteins; D007210:Indoleacetic Acids; D009097:Multienzyme Complexes; C030737:indoleacetic acid; D004247:DNA