Abstract
Beyond their role as protein-building units, amino acids are modulators of multiple behaviours in different microorganisms. In the root-colonizing beneficial bacterium Pseudomonas putida (recently proposed to be reclassified as alloputida) KT2440, current evidence suggests that arginine functions both as a metabolic indicator and as an environmental signal molecule, modulating processes such as chemotactic responses, siderophore-mediated iron uptake or the levels of the intracellular second messenger cyclic diguanylate (c-di-GMP). Using microcalorimetry and extracellular flux analysis, in this work we have studied the metabolic adaptation of P. putida KT2440 to the presence of L-arginine in the growth medium, and the influence of mutations related to arginine metabolism. Arginine causes rapid changes in the respiratory activity of P. putida, particularly magnified in a mutant lacking the transcriptional regulator ArgR. The metabolic activity of mutants affected in arginine transport and metabolism is also altered during biofilm formation in the presence of the amino acid. The results obtained here further support the role of arginine as a metabolic signal in P. putida and the relevance of ArgR in the adaptation to the amino acid. They also serve as proof of concept on the use of calorimetric and extracellular flux techniques to analyse metabolic responses in bacteria and the impact of different mutant backgrounds on such responses.
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Abbreviations
- ADC pathway:
-
arginine decarboxylase pathway
- ADH pathway:
-
arginine dehydrogenase pathway
- ADI pathway:
-
arginine deiminase pathway
- AST pathway:
-
arginine succinyltransferase pathway
- Seahorse:
-
extracellular flux analyser XFe96 Seahorse (Agilent)
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Author Contributions
CSR carried out the Seahorse experiments and growth under different nutrients, MAH carried out microbiology and calorimetric assays, MEU and SR conceptualized and supervised the experiments and wrote the manuscript.
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This article does not contain any studies with human participants or animals performed by any of the authors.
Funding
MEU was granted from the European Union’s Horizon 2020 Research and Innovation Programme under grant agreement N° 101004806 MOSBRI, DSB-UROM TNA access provider for Seahorse experiment. Financial support from the Sapienza University of Rome (RM120172A7AD98EB to S.R. and AR12117A63EE6037 to C.S.R.), and grant PID2019-109372GB-I00 funded by MCIN/AEI/10.13039/501100011033 (M.E.U. and M.A.M.H.), are also acknowledged.
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Scribani-Rossi, C., Molina-Henares, M.A., Espinosa-Urgel, M., Rinaldo, S. (2024). Exploring the Metabolic Response of Pseudomonas putida to L-arginine. In: Advances in Experimental Medicine and Biology(). Springer, Cham. https://doi.org/10.1007/5584_2024_797
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DOI: https://doi.org/10.1007/5584_2024_797
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