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Cellular needs and conditions vary from cell to cell and change within individual cells over time. For example, the required enzymes and energetic demands of stomach cells are different from those of fat storage cells, skin cells, blood cells, and nerve cells. Furthermore, a digestive cell works much harder to process and break down nutrients during the time that closely follows a meal compared with many hours after a meal. As these cellular demands and conditions vary, so do the amounts and...
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The operon model represents a fundamental mechanism of gene regulation in prokaryotes, enabling coordinated expression of genes involved in related metabolic or functional pathways. Operons consist of structural genes, a promoter, and an operator, with transcription regulated by repressors, activators, and small effector molecules.Structure and Function of OperonsAn operon is a cluster of structural genes transcribed together under the control of a single promoter. The promoter region...
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Lipids also are sources of energy that power cellular processes. Like carbohydrates, lipids are composed of carbon, hydrogen, and oxygen, but these atoms are arranged differently. Most lipids are nonpolar and hydrophobic. Major types include fats and oils, waxes, phospholipids, and steroids.
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Applied evolution: Dual dynamic regulations-based approaches in engineering intracellular malonyl-CoA availability.

Junjun Wu1, Lin Zhou1, Xuguo Duan2

  • 1College of Food Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu, 210095, China.

Metabolic Engineering
|August 19, 2021
PubMed
Summary

Engineered microbial pathways using evolution and dual dynamic regulations significantly boosted malonyl-CoA availability. This breakthrough enables scalable, autonomous production of valuable compounds like (2S)-naringenin.

Keywords:
Pathway optimizationPhenylpropanoidsPlant natural productsStilbeneSynthetic biology

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Area of Science:

  • Synthetic biology
  • Metabolic engineering
  • Microbial biotechnology

Background:

  • Malonyl-CoA is crucial for synthesizing valuable compounds but its availability is tightly regulated in cells.
  • Existing microbial production methods are often limited by inefficient malonyl-CoA supply.
  • Novel strategies are needed to overcome cellular constraints and enhance malonyl-CoA production.

Purpose of the Study:

  • To develop a robust and dynamic system for controlling malonyl-CoA availability in microbes.
  • To engineer a microbial platform for enhanced production of malonyl-CoA derived compounds.
  • To create an autonomous fermentation process minimizing human intervention.

Main Methods:

  • Genome mining and phylogenomic analysis identified highly active acetyl-CoA carboxylase (ACC) families from Salmonella enterica.
  • Designed orthogonal, bi-functional quorum-sensing (QS)-based regulatory tools coupled with T7 RNA polymerase for dual dynamic gene control.
  • Integrated QS circuits with Salmonella ACC and CRISPR interference (CRISPRi) to reprogram central metabolism and malonyl-CoA pathway.

Main Results:

  • Achieved a 29-fold increase in malonyl-CoA availability by rewiring the metabolic pathway.
  • Successfully synthesized (2S)-naringenin with a record production of 1073.8 mg/L, while reducing by-products.
  • Demonstrated autonomous fermentation, eliminating the need for external inducers and continuous supervision.

Conclusions:

  • The developed 'evolution: dual dynamic regulations' approach provides a scalable and economically viable method for microbial production of malonyl-CoA derived compounds.
  • This strategy overcomes inherent cellular limitations, paving the way for efficient biosynthesis of complex molecules.
  • The autonomous fermentation system represents a significant advancement in bioprocess engineering.