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Sci Rep ; 6: 26065, 2016 05 17.
Artigo em Inglês | MEDLINE | ID: mdl-27184291

RESUMO

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.


Assuntos
Vias Biossintéticas/genética , Escherichia coli/enzimologia , Escherichia coli/genética , Ácidos Indolacéticos/metabolismo , Engenharia Metabólica , Complexos Multienzimáticos/genética , Complexos Multienzimáticos/metabolismo , DNA/metabolismo , Enzimas/metabolismo , Escherichia coli/metabolismo , Proteínas de Escherichia coli/metabolismo , Ligação Proteica
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