Functional γ-Aminobutyrate Shunt in Listeria monocytogenes: role in acid tolerance and succinate biosynthesis.
Listeria monocytogenes, the causative agent of human listeriosis, is known for its ability to withstand severe environmental stresses. The glutamate decarboxylase (GAD) system is one of the principal systems utilized by the bacterium to cope with acid stress, a reaction that produces γ-aminobutyrate (GABA) from glutamate. Recently, we have shown that GABA can accumulate intracellularly under acidic conditions, even under conditions where no extracellular glutamate-GABA exchange is detectable. The GABA shunt, a pathway that metabolizes GABA to succinate, has been described for several other bacterial genera, and the present study sought to determine whether L. monocytogenes has this metabolic capacity, which, if present, could provide a possible route for succinate biosynthesis in L. monocytogenes. Using crude protein extracts from L. monocytogenes EGD-e, we show that this strain exhibits activity for the two main enzyme reactions in the GABA shunt, GABA aminotransferase (GABA-AT) and succinic semialdehyde dehydrogenase (SSDH). Two genes were identified as candidates for encoding these enzyme activities, argD (GABA-AT) and lmo0913 (SSDH). Crude protein extracts prepared from a mutant lacking a functional argD gene significantly reduced GABA-AT activity, while an lmo0913 mutant lost all detectable SSDH activity. The deletion of lmo0913 increased the acid tolerance of EGD-e and showed an increased accumulation of intracellular GABA, suggesting that this pathway plays a significant role in the survival of this pathogen under acidic conditions. This is the first report of such a pathway in the genus Listeria, which highlights an important link between metabolism and acid tolerance and also presents a possible compensatory pathway to partially overcome the incomplete tricarboxylic acid cycle of Listeria.
Bacterial Stress Response Group, Microbiology, School of Natural Sciences, National University of Ireland, Galway, Galway, Ireland.
Conor
C
Feehily
Conor P
CP
O'Byrne
Kimon Andreas G
KA
Karatzas
D000613:Aminobutyrates / Q000378:metabolism*; D002264:Carboxylic Acids / Q000378:metabolism*; D018698:Glutamic Acid / Q000378:metabolism; D006801:Humans; D008089:Listeria monocytogenes / Q000378:metabolism*; D053858:Metabolic Networks and Pathways / Q000235:genetics*; D019802:Succinic Acid / Q000378:metabolism*
D000613:Aminobutyrates; D002264:Carboxylic Acids; D018698:Glutamic Acid; D019802:Succinic Acid