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Published on: January 18, 2014
CLCF, Not CrcB, Protects Pseudomonas putida ATCC 12633 During Intense Fluoride Stress
Anthony G Dodge1,2, Madeline R O'Connor1,3, Lawrence P Wackett1,2,3
1Biotechnology Institute, University of Minnesota, St. Paul, Minnesota, USA.
Environmental Microbiology
|July 17, 2026
Summary
Engineered bacteria expressing CLCF transporters achieved high resistance to sodium fluoride and degraded organofluorine compounds. This advance is key for biodegrading fluorinated chemicals and biosynthesis.
Area of Science:
- Microbiology
- Biotechnology
- Environmental Science
Background:
- Fluoride is abundant but toxic to cells, necessitating microbial export mechanisms.
- Prokaryotes utilize either CrcB (fluoride export channels) or CLCF (fluoride/proton antiporters), but not both.
Purpose of the Study:
- To engineer Pseudomonas putida for enhanced fluoride tolerance and organofluorine compound biodegradation.
- To investigate the role of CLCF transporters in high-concentration fluoride environments.
Main Methods:
- Pseudomonas putida engineered with CLCF exporter gene.
- Adaptive evolution under high sodium fluoride conditions.
- Genetic analysis of evolved strains, including CrcB knockout confirmation.
Main Results:
- Engineered cells achieved resistance to >500 mM sodium fluoride.
- Degradation of 150 mM 2-fluoropropionic acid and export of 150 mM fluoride achieved.
- Adaptive evolution uniformly increased CLCF gene copy and mutated native CrcB.
Conclusions:
- The CLCF antiporter is crucial for biodegrading high concentrations of organofluorine compounds.
- Sustaining microbial viability at high fluoride levels enables biodegradation of fluorinated chemicals.
- This research supports engineering prokaryotes for industrial applications in fluorochemicals and biosynthesis.
