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Benzoate transport in Pseudomonas putida CSV86
Alpa Choudhary1, Hemant Purohit2, Prashant S Phale1
1Department of Biosciences and Bioengineering, Indian Institute of Technology-Bombay, Powai, Mumbai 400076, India.
FEMS Microbiology Letters
|June 8, 2017
Summary
Pseudomonas putida CSV86 utilizes aromatic compounds via specific transporters. Benzoate transport is an inducible, active process driven by proton motive force, involving outer and inner membrane proteins.
Area of Science:
- Microbial metabolism
- Bacterial transport systems
- Pseudomonas genomics
Background:
- Pseudomonas putida CSV86 metabolizes diverse aromatic compounds.
- Genome analysis identified putative transporters for various aromatic acids.
Purpose of the Study:
- Investigate the molecular mechanisms of benzoate transport in P. putida CSV86.
- Characterize the benzoate transport system at the genetic and biochemical levels.
Main Methods:
- Bioinformatic analysis of the ben locus (benK-catA-benE-benF).
- Protein topology prediction for transporters BenK and BenE.
- Biochemical assays to determine transport kinetics and energy dependence.
- Transcriptional analysis of benzoate and glucose-grown cells.
Main Results:
- Identified benzoate transport and metabolism genes clustered at the ben locus.
- Predicted transmembrane helices for BenK (12) and BenE (11).
- Benzoate transport is inducible, active, proton motive force-dependent, with high affinity (Km 4.5 μM).
- Inducible expression of benF, benK, and benE suggests roles for both porin and inner membrane transporters.
Conclusions:
- P. putida CSV86 employs a multi-component system for benzoate uptake.
- Outer membrane porin (BenF/OprD) and inner membrane transporters (BenK, BenE) are crucial for benzoate transport.
- The transport process is energy-dependent, utilizing proton motive force.
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