In Vitro and In Silico Assessment of the Probiotic and Technological Potential of Lacticaseibacillus rhamnosus MW019593 Isolated from Algerian Cow’s Milk

Abstract

This study evaluated the in vitro and in silico probiotic potential and technological characteristics of Lacticaseibacillus rhamnosus MW019593 isolated from Algerian cow’s milk. Safety was verified by the absence of hemolytic activity, biogenic amine production, and antibiotic sensitivity. Functional probiotic properties of L. rhamnosus MW019593 included moderate inhibition of gastrointestinal pathogens such as Salmonella, Listeria, Clostridioides, and Escherichia coli, tolerance to pH 2–3 and up to 1% bile salts, 46.4% hydrophobicity, 42.4% autoaggregation, and 12.9% adhesion to Caco-2/TC7 cells with out cytotoxicity. The ability to form biofilms with a thickness of 23.7 µm, comparable to L. rhamnosus ATCC 53103, was observed using confocal microscopy. Technologically, L. rhamnosus MW019593 and ATCC 53103 showed similar growth profiles and biomass yields, reaching approximately 5× 109 CFU/mL with a 63% yield after lyophilization. No significant difference in strain viability was noted at 4 °C and−20 °C, which are suitable temperatures to maintain their viability for 3 months. Whole genome sequencing (WGS) was conducted on L. rhamnosus MW019593, followed by comprehensive in silico genomic analysis. Taxonomic validation using average nucleotide identity (ANI) and genome-to-genome distance hybridization (GGDH) confirmed the strain’s affiliation to this species. Gene screening revealed the absence of virulence and antibiotic resistance genes, while beneficial genes, including those for bacteriocin production, were identified. These findings, supported by the demonstrated safety, functional, and technological properties of the strain, highlight its promising potential for food and therapeutic applications, pending in vivo validation.