Engineering cell morphology by CRISPR interference in Acinetobacter baylyi ADP1

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.