Selection of GABA-Producing Lactic Acid Bacteria Strains by Polymerase Chain Reaction Using Novel gadB and gadC...

Gamma-aminobutyric acid (GABA) has been attributed to health-promoting properties and has received attention from the food industry as an attractive bioactive compound for the development of functional foods. Some lactic acid bacteria (LAB) produce GABA through a glutamate decarboxylase encoded by gadB and a glutamate/GABA antiporter encoded by gadC. In this study, we develop a molecular screening method based on a polymerase chain reaction able to detect those genes in different LAB species through the use of novel multispecies primers. PCR was performed in 92 LAB strains of six different species. The primer pair designed for gadB allowed its identification in Lactiplantibacillus plantarum, Lactococcus cremoris, Lactococcus lactis, Levilactobacillus brevis, Limosilactobacillus fermentum, and Limosilactobacillus reuteri strains. For gadC, two different primer pairs were designed for its detection in different species. Glutamate decarboxylase activity (GAD assay) and GABase enzymatic quantification were also assessed. Among those strains showing glutamate decarboxylase activity, 93.2% harbored the gadB gene, and those showing GABA production had the gadB gene and exhibited glutamate decarboxylase activity. PCR detection of gadB correlates strongly with GABA production and constitutes a good strategy for the selection of LAB with high yields (>18 mM) that could be used for the development of GABA-enriched functional foods.