Biodiversity andγ-Aminobutyric Acid Production by Lactic Acid Bacteria Isolated from Traditional Alpine Raw Cow’s Milk Cheeses

Abstract

“Nostrano-cheeses” are traditional alpine cheeses made from raw cow’s milk in Trentino-Alto Adige, Italy. This study identified lactic acid bacteria (LAB) developing during maturation of “Nostrano-cheeses” and evaluated their potential to produceγ-aminobutyric acid (GABA), an immunologically active compound and neurotransmitter. Cheese samples were collected on six cheese-making days, in three dairy factories located in different areas of Trentino and at different stages of cheese ripening (24 h, 15 days, and 1, 2, 3, 6, and 8 months). A total of 1,059 LAB isolates were screened using Random Amplified Polymorphic DNA-PCR (RAPD-PCR) and differentiated into 583 clusters. LAB strains from dominant clusters (n=97) were genetically identified to species level by partial 16S rRNA gene sequencing. LAB species most frequently isolated wereLactobacillus paracasei,Streptococcus thermophilus, andLeuconostoc mesenteroides. The 97 dominant clusters were also characterized for their ability in producing GABA by high-performance liquid chromatography (HPLC). About 71% of the dominant bacteria clusters evolving during cheeses ripening were able to produce GABA. Most GABA producers wereLactobacillus paracaseibut other GABA producing species includedLactococcus lactis,Lactobacillus plantarum,Lactobacillus rhamnosus,Pediococcus pentosaceus, andStreptococcus thermophilus. NoEnterococcus faecalisorSc. macedonicusisolates produced GABA. The isolate producing the highest amount of GABA (80.0±2.7 mg/kg) was aSc. thermophilus.