Osmoprotection by Pipecolic Acid in Sinorhizobium meliloti: Specific Effects of d and l Isomers

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American Society for Microbiology

RESUMO

dl-Pipecolic acid (dl-PIP) promotes growth restoration of Sinorhizobium meliloti cells facing inhibitory hyperosmolarity. Surprisingly, d and l isomers of this imino acid supplied separately were not effective. The uptake of l-PIP was significantly favored in the presence of the d isomer and by a hyperosmotic stress. Chromatographic analysis of the intracellular solutes showed that stressed cells did not accumulate radiolabeled l-PIP. Rather, it participates in the synthesis of the main endogenous osmolytes (glutamate and the dipeptide N-acetylglutaminylglutamine amide) during the lag phase, thus providing a means for the stressed cells to recover the osmotic balance. 13C nuclear magnetic resonance analysis was used to determine the fate of d-PIP taken into the cells. In the absence of l-PIP, the imported d isomer was readily degraded. Supplied together with its l isomer, d-PIP was accumulated temporarily and thus might contribute together with the endogenous osmolytes to enhance the internal osmotic strength. Furthermore, it started to disappear from the cytosol when the l isomer was no longer available in the culture medium (during the late exponential phase of growth). Together, these results show an uncommon mechanism of protection of osmotically stressed cells of S. meliloti. It was proved, for the first time, that the presence of the two isomers of the same molecule is necessary for it to manifest an osmoprotective activity. Indeed, d-PIP seems to play a major role in cellular osmoadaptation through both its own accumulation and improvement of the utilization of the l isomer as an immediate precursor of endogenous osmolytes.

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