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Updated: Aug 10, 2025

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Developing a genetic engineering method for Acetobacterium wieringae to expand one-carbon valorization pathways.

João P C Moreira1,2, John T Heap3, Joana I Alves1,2

  • 1CEB - Centre of Biological Engineering, University of Minho, 4710-057, Braga, Portugal.

Biotechnology for Biofuels and Bioproducts
|February 15, 2023
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Summary

Researchers developed a new electrotransformation protocol for Acetobacterium wieringae, enabling genetic modification. This breakthrough allows for non-native acetone production in these bacteria, advancing sustainable chemical manufacturing.

Keywords:
AcetobacteriumAcetogenBiofuelsCarbon monoxideChemicalsElectroporationGenetic engineeringSyngasTransformation

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Area of Science:

  • Microbiology
  • Synthetic Biology
  • Biotechnology

Background:

  • Developing cost-effective bioprocesses is crucial for replacing fossil fuels.
  • Utilizing low-cost gas streams (CO, CO2, H2) for chemical production offers a sustainable alternative.
  • Non-model autotrophic acetogens like Acetobacterium wieringae hold potential for industrial applications but require genetic tools.

Purpose of the Study:

  • To develop and optimize an electrotransformation protocol for Acetobacterium wieringae.
  • To establish genetic tools for manipulating A. wieringae for industrial bioprocessing.
  • To demonstrate the feasibility of introducing non-native metabolic pathways into A. wieringae.

Main Methods:

  • Optimized electrotransformation protocol for Acetobacterium wieringae strains JM, DSM 1911, and A. woodii DSM 1030.
  • Validated thiamphenicol resistance marker (catP) and various plasmid origins of replication (pBP1, pCB102, pCD6, pIM13).
  • Introduced the acetone production operon from Clostridium acetobutylicum into Acetobacterium spp.

Main Results:

  • Achieved high electrotransformation efficiencies (up to 2.0 × 10^3 CFU/μgDNA) in A. wieringae strains.
  • Identified key factors influencing electrotransformation efficiency, including cell density, buffer pH, electric field strength, and plasmid concentration.
  • Successfully engineered Acetobacterium spp. for non-native acetone production via plasmid-based expression.

Conclusions:

  • Established an efficient electrotransformation method for biotechnologically relevant Acetobacterium strains.
  • Unlocked genetic and metabolic engineering capabilities for A. wieringae.
  • Demonstrated the first instance of non-native acetone production in A. wieringae.