Postbiotic Potential of Newly Isolated Riboflavin-Overproducing Lactiplantibacillus plantarum Strains

Lactic acid bacteria (LAB) are food-grade microorganisms able to produce and release bioactive compounds of interest to human health. Some LAB strains can synthesize vitamin B2, i.e., riboflavin, a micronutrient essential for cellular metabolism. In this work, six Lactiplantibacillus plantarum isolated from fruits of the Mediterranean area were exposed to the selective pressure of roseoflavin in order to select spontaneous riboflavin-overproducing phenotypes. The best strains, as determined by the level of riboflavin produced, were characterized for some basic probiotic features, including antibacterial activity, production of organic acids, antibiotic resistance, and survival under digestive stresses using an in vitro gut model. The strain L. plantarum Lp 187_B2, which produced the highest riboflavin level (6 mg/L), exhibited good resistance to gastro-intestinal stress and a relevant capacity to antagonize undesired bacteria, was selected for additional investigations to assess its capacity to protect intestinal homeostasis. When used as a postbiotic, Lp 187_B2 significantly increased trans-epithelial electrical resistance (TEER) in Caco-2 cell monolayers, an in vitro model of the intestinal barrier. Moreover, in a Caco-2/THP-1 co-culture system, mimicking the inflamed bowel, Lp 187_B2 postbiotics significantly inhibited the release of TNF-alpha by macrophages, thus pointing to gut-barrier strengthening and potential anti-inflammatory properties. Though a validation in vivo is required, our preliminary results indicate that L. plantarum Lp 187_B2 could be successfully applied as both probiotic and postbiotic formulations for improving human health.