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Engineering cell morphology by CRISPR interference in Acinetobacter baylyi ADP1.

Jin Luo1, Elena Efimova1, Daniel Christoph Volke2

  • 1Faculty of Engineering and Natural Sciences, Tampere University, Tampere, Finland.

Microbial Biotechnology
|August 25, 2022
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Summary

Engineered bacteria using CRISPR interference (CRISPRi) produced more wax esters by redirecting carbon flow and increasing cell size. This demonstrates cell morphology engineering for enhanced microbial lipid production.

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Area of Science:

  • Synthetic biology
  • Metabolic engineering
  • Microbial biotechnology

Background:

  • Microbial production of intracellular compounds can be enhanced by optimizing carbon flux and cell size.
  • Clustered regularly interspaced short palindromic repeats interference (CRISPRi) offers precise gene expression control for metabolic engineering.

Purpose of the Study:

  • To engineer Acinetobacter baylyi ADP1 for increased intracellular wax ester production using CRISPRi.
  • To investigate the impact of cell morphology modification on lipid synthesis.

Main Methods:

  • Established an inducible CRISPRi system in Acinetobacter baylyi ADP1 for controlled gene repression.
  • Targeted the glyoxylate shunt to redirect carbon flux towards wax ester synthesis.
  • Utilized CRISPRi to repress the essential ftsZ gene, altering cell division and morphology.

Main Results:

  • Successfully engineered Acinetobacter baylyi ADP1 with a tunable CRISPRi system.
  • Redirected carbon flux towards wax esters by targeting the glyoxylate shunt.
  • Generated enlarged filamentous and spherical cells by repressing ftsZ, leading to increased wax ester production metrics.

Conclusions:

  • CRISPRi is an effective tool for engineering Acinetobacter baylyi for enhanced intracellular lipid production.
  • Cell morphology engineering, achieved through ftsZ repression, significantly boosts wax ester yields.
  • This study highlights the potential of combining metabolic and morphological engineering for improved microbial bioproduction.