We investigated the potential probiotic properties of Limosilactobacillus fermentum KGC1601 isolated from Panax ginseng. Ginseng cultivated in an experimental field of the Korea Ginseng Research Institute was fermented, followed by single colony selection from MRS agar. We performed 16s-rRNA sequencing and whole-genome analysis to identify L. fermentum and evaluate the biosafety parameters of this strain, respectively. We confirmed this strain was susceptible to six antibiotics, as proposed by the European Food Safety Authority, did not produce biogenic amines, and did not exhibit any hemolytic activity. Acid resistance and bile salt tolerance, which are essential properties of a probiotic agent, were investigated. Notably, distinguishing properties of this strain were that it exhibited excellent bile salt tolerance and anti-inflammatory effects. The excellent bile salt tolerance was confirmed by scanning electron microscopy. Through qRT-PCR and ELISA studies, it was revealed that L. fermentum KGC1601 pre-treatment up-regulates anti-inflammatory cytokines and down-regulates pro-inflammatory cytokines in RAW 264.7 cells. Consequently, we suggested that L. fermentum KGC1601 can be safely used as a potential anti-inflammatory functional probiotic agent.

In this study, we determined the probiotic properties and safety of Limosilactobacillus reuteri KGC1901 isolated from Korean Panax ginseng root. This strain was identified based on its 16s rRNA sequence, and the absence of genes related to antibiotic resistance and virulence was confirmed through whole genome analysis in terms of safety. Moreover, this strain had no antibiotic resistance to eight antibiotics as proposed by the European Food Safety Authority, did not show hemolytic activity on blood agar, and did not produce biogenic amines. L. reuteri KGC1901 also showed the capability to survive at low pH and in presence of bile salts and sufficiently adhered to HT-29 cells and mucin. The adhesion ability to HT-29 was confirmed by immunofluorescence staining. These results indicated the excellent viability of L. reuteri KGC1901 in the human gastrointestinal tract. Additionally, L. reuteri KGC1901 had antimicrobial activity against Clostridium difficile, and the presence of the reuterin (3-HPA) biosynthetic gene cluster in the genome was revealed. Furthermore, the cell-free supernatant of KGC1901 had DPPH scavenging activity and reduced the nitric oxide production in LPS-stimulated RAW 264.7 cells. Based on these results, it was confirmed that L. reuteri KGC1901 derived from ginseng has sufficient potential to be used as a probiotic.