L-Alanine Exporter, AlaE, of Escherichia coli Functions as a Safety Valve to Enhance Survival under Feast Conditions
Abstract
:1. Introduction
2. Results and Discussion
2.1. Susceptibility of MG1655 and Its Derivatives to L-Alanyl-L-Alanine (Ala-Ala)
2.2. Accumulation of Intracellular Alanine in MG1655 and its Derivatives
2.3. Coculture Assay in the Presence of Ala-Ala
3. Materials and Methods
3.1. Bacterial Strains, Plasmids, and Culture Conditions
3.2. Construction of a Single Mutant Lacking dadA and Double Mutants Lacking both alaE and dadA
3.3. Susceptibility of the Mutants to L-alanyl-L-alanine (Ala-Ala)
3.4. Determination of Intracellular Amino Acid Concentration
3.5. Coculture Assay
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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MIC (mg mL −1) | ||
---|---|---|
Strains | Glucose | Glycerol |
MG1655 | >1.25 | >1.25 |
MG1655∆dadA | >1.25 | >1.25 |
MG1655∆alaE | 0.04 | 0.16 |
MG1655∆dadA∆alaE | 0.0025 | 0.0025 |
MLA301 | >1.25 | >1.25 |
MLA301∆alaE | 0.0025 | 0.0025 |
Strain or Plasmid | Characteristics | Reference |
---|---|---|
Strain | ||
E. coli MG1655 | Wild type | Laboratory strain |
E. coli JW1178 | dadA disruptant derived from BW25113, KMr | [34] |
E. coli JW2645 | alaE disruptant derived from BW25113, KMr | [34] |
E. coli MG1655 alaE | alaE disruptant derived from MG1655, KMr | [29] |
E. coli MG1655 dadA | dadA disruptant derived from MG1655, KMr | This study |
E. coli MG1655 dadA alaE | MG1655 dadA derivative with a deletion in the alaE gene, KMr | This study |
E. coli MLA301 | MG1655 alr::FRT, dadX::FRT, yfdZ::FRT, avtA::GM, yfbQ::KM | [20] |
E. coli MLA301 alaE | MLA301 derivative with a deletion in the alaE gene, GMr, KMr | [27] |
Plasmid | ||
pCP20 | FLP+, λcI857−, λpRRepts, Ampr, CPr | [35] |
pKD46 | Red recombinase expression plasmid | [36] |
pBR322 | Ampr, TETr, cloning vector | [27] |
pBR325 | Ampr, TETr, CPr, cloning vector | Laboratory stock |
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Katsube, S.; Ando, T.; Yoneyama, H. L-Alanine Exporter, AlaE, of Escherichia coli Functions as a Safety Valve to Enhance Survival under Feast Conditions. Int. J. Mol. Sci. 2019, 20, 4942. https://doi.org/10.3390/ijms20194942
Katsube S, Ando T, Yoneyama H. L-Alanine Exporter, AlaE, of Escherichia coli Functions as a Safety Valve to Enhance Survival under Feast Conditions. International Journal of Molecular Sciences. 2019; 20(19):4942. https://doi.org/10.3390/ijms20194942
Chicago/Turabian StyleKatsube, Satoshi, Tasuke Ando, and Hiroshi Yoneyama. 2019. "L-Alanine Exporter, AlaE, of Escherichia coli Functions as a Safety Valve to Enhance Survival under Feast Conditions" International Journal of Molecular Sciences 20, no. 19: 4942. https://doi.org/10.3390/ijms20194942