IntroductionProbioticLactiplantibacillus plantarumMC5 produces large amounts of exopolysaccharides (EPS), and its use as a compound fermentor can greatly improve the quality of fermented milk.MethodsTo gain insight into the genomic characteristics of probiotic MC5 and reveal the relationship between its EPS biosynthetic phenotype and genotype, we analyzed the carbohydrate metabolic capacity, nucleotide sugar formation pathways, and EPS biosynthesis-related gene clusters of strain MC5 based on its whole genome sequence. Finally, we performed validation tests on the monosaccharides and disaccharides that strain MC5 may metabolize.ResultsGenomic analysis showed that MC5 has seven nucleotide sugar biosynthesis pathways and 11 sugar-specific phosphate transport systems, suggesting that the strain can metabolize mannose, fructose, sucrose, cellobiose, glucose, lactose, and galactose. Validation results showed that strain MC5 can metabolize these seven sugars and produce significant amounts of EPS (> 250 mg/L). In addition, strain MC5 possesses two typicalepsbiosynthesis gene clusters, which include the conserved genesepsABCDE,wzx, andwzy, six key genes for polysaccharide biosynthesis, and one MC5-specificepsGgene.DiscussionThese insights into the mechanism of EPS-MC5 biosynthesis can be used to promote the production of EPS through genetic engineering.