Safety Assessment of Lactiplantibacillus plantarum TWK10 Based on Whole-Genome Sequencing, Phenotypic, and Oral Toxicity Analysis

Abstract

Lactiplantibacillus plantarum TWK10 (TWK10), isolated from Taiwanese pickled cabbage, has been demonstrated to exert beneficial probiotic effects in both mice and humans. Here, we comprehensively assessed the safety of TWK10 using both in vivo and in vitro approaches, including whole-genome sequence analysis, an assessment of hemolytic activity, and performing an antimicrobial susceptibility test, the Ames bacterial reverse mutation assay, the chromosomal aberration test, a rodent peripheral blood micronucleus test, and the 28-day subacute oral toxicity assay. The results showed that there was no significant increase in the incidence of reverse mutations or chromosomal aberrations following exposure to TWK10. Moreover, no significant changes were detected either in the number of reticulocytes or the incidence of micronuclei in ICR mice, and no subacute toxicity was recorded in SD rats at the oral TWK10 dosage of 2000 mg/kg body weight/day repeated for 28 days. Additionally, TWK10 exhibited no hemolytic activity and was susceptible to all the antibiotics tested, except kanamycin. However, no antimicrobial resistance genes, virulence factors, or genes involved in biogenic amine synthesis were found in the genome of TWK10. Our findings demonstrated that TWK10 has high potential of being safe for human consumption as a probiotic.

Effects of heat‐killed Lactiplantibacillus plantarumTWK10 on exercise performance, fatigue, and muscle growth in healthy male adults

“…2.1 | Preparation of L. plantarum TWK10 TWK10 was isolated from Taiwanese pickled cabbage, "Pao-tsai" (Chen et al, 2013), and identified as Lactiplantibacillus plantarum subsp. plantarum (Hsu et al, 2022). Heat-killed TWK10 cells were prepared by SYNBIO TECH INC. (Kaohsiung, Taiwan) in capsule form with a dose of 1.5 × 10 10 cells (corresponding to 1.5 × 10 10 colony-forming units) standardized with microcrystalline cellulose and maltodextrin.…”

Investigate the impact of probiotics of lactic acid bacteria obtained from various local sources on some pathogenic bacteria

“…Among these, the genus Lactobacilli stands out as the largest due to its diverse species. Because the species within this genus are recognized for their safety and numerous health-promoting qualities, several strains have garnered considerable attention (8).…”

Special Issue: Beneficial Properties and Safety of Lactic Acid Bacteria

“…However, as aforementioned, safety is a strain-based characteristic and thus cannot be associated with an entire species. Hsu et al [9] reported on Lb. plantarum TWK10, isolated from Taiwanese pickled cabbage; in addition to its suggested probiotic properties, as evaluated in mice and humans, the authors assessed this strain's safety via in vitro and in vivo experiments.…”

“…The novel tools utilized in the evaluation of the safety of microbial cultures, including DNA-associated experimental approaches, have become routine in the last two decades. Considering this, the validation of safety, both of new microbial and currently applied cultures, is now considered In addition to "classical" PCR-based approaches, whole genome sequencing and the appropriate analysis of the generated data have become routine in the evaluation of the safety profile of microbial cultures [7][8][9].…”

“…The safety profiles of strains of enterococci, Lactiplantibacillus plantarum and Weissella cibaria were assessed in the research of Tsanasidou et al [7], Jang et al [8] and Hsu et al [9]. These strains were shown to have various beneficial features, ranging from their production of bacteriocins, to their favorable probiotic properties.…”