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A self-regulated network for dynamically balancing multiple precursors in complex biosynthetic pathways.

Yusong Zou1, Jianli Zhang1, Jian Wang1

  • 1School of Chemical, Materials and Biomedical Engineering, College of Engineering, The University of Georgia, Athens, GA, 30602, USA.

Metabolic Engineering
|February 5, 2024
PubMed
Summary

This study introduces a self-regulated microbial network to manage competing metabolic pathways. This dynamic balancing enhances the production of complex molecules like 4-hydroxycoumarin (4-HC) by optimizing precursor flow.

Keywords:
4-HydroxycoumarinBiosynthesisDynamic regulationGenetic biosensorsMultiple precursors

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Area of Science:

  • Synthetic Biology
  • Metabolic Engineering
  • Biochemical Engineering

Background:

  • Microbial synthesis offers a sustainable alternative to traditional methods but faces challenges with competing metabolic pathways.
  • Competition for cellular resources and carbon flux conflicts can reduce the efficiency and yield of target product biosynthesis.
  • Complex synthetic pathways requiring multiple precursors from shared metabolic nodes exacerbate these production limitations.

Purpose of the Study:

  • To develop a self-regulated network to mitigate precursor competition in complex microbial synthetic pathways.
  • To dynamically control metabolic flux and gene expression for improved production efficiency.
  • To demonstrate the efficacy of this approach using the 4-hydroxycoumarin (4-HC) synthetic pathway as a model system.

Main Methods:

  • Designed a self-regulated network utilizing an intermediate metabolite as a trigger.
  • Dynamically rewired metabolic flux of pyruvate and controlled gene expression in the 4-HC pathway.
  • Employed transcriptomic analysis to monitor gene expression changes in response to intermediate concentrations.

Main Results:

  • Successfully achieved self-regulation of multiple precursors in the 4-HC synthetic pathway.
  • Demonstrated enhanced production titers of 4-HC through dynamic metabolic flux control.
  • Transcriptomic data confirmed dynamic changes in pyruvate kinase (PykF) and SdgA gene expression correlating with intermediate levels.

Conclusions:

  • Established a self-regulated network capable of dynamically balancing competing metabolic flux for precursor biosynthesis.
  • The developed system provides a novel strategy for optimizing microbial production of compounds requiring complex metabolic pathways.
  • Offers insights into managing precursor interference and competition in microbial biosynthesis for improved yields.