The glutamate decarboxylase (GAD) system is critical to the survival of Listeria monocytogenes LO28 at low-pH stress (<pH 4.0). The GAD system classically involves two proteins, a glutamate decarboxylase enzyme coupled to a glutamate/gamma-aminobutyrate antiporter, which results in the consumption of an intracellular proton for each glutamate entering the system. Uniquely among prokaryotes, some strains of L. monocytogenes, including strain LO28, possess genes encoding three decarboxylases (gadD1, gadD2, and gadD3) and two antiporters (gadT1 and gadT2). These are organized in two pairs (gadD1T1 and gadD2T2) and a distinct gadD3. While the creation of a gadD3 mutant has not been possible, analysis of 15 isogenic mutants has confirmed previous observations that GadD2/T2 are primarily responsible for surviving severe acid challenge (pH 2.8). However, we have now established that GadD1 plays a major role in growth at mildly acidic pHs (pH 5.1). When strain variation studies revealed that a large number of L. monocytogenes strains (including all serotype 4 strains) lack the gadD1 gadT1 pair, low-pH growth assays were carried out. It was found that the majority of strains that grew poorly at pH 5.1 lacked these genes. The strain-variable ability to grow in mildly acidic conditions may explain why non-serotype 4 strains of L. monocytogenes predominate in foods.