An optimal culture medium and highly stable biometabolites are important in industrial production processes. The response surface methodology with a Box–Behnken design was performed to determine the optimal culture medium of an engineered Aspergillus oryzae strain for cordycepin production by submerged fermentation. The influences of glucose, yeast extract, and adenine concentrations on cordycepin production were explored, and their concentrations were used for experimental design. The results reveal that the optimal culture components involved 30.0 g/L of glucose, 9.8 g/L of yeast extract, and 1.5 g/L of adenine. As predicted, the maximum cordycepin concentration (1724.53 ± 18.30 mg/L) was obtained with a short fermentation time (2 days). A significant increase in cordycepin yield (>50% increase) was observed in the culture grown in the optimized culture medium compared to that grown in the basal medium. A xanthine oxidase inhibitory activity assay demonstrated that the cordycepin product had a pharmacological function. It exhibited strong stability under high thermal and acidic conditions, with over 95% product recovery. The findings of this study are valuable for developing cost-effective processes for producing health-benefiting products.

Lactiplantibacillus species have been historically used for food applications. Although several species are regarded as safe according to their regulatory status, the safety issues and functional roles of these lactic acid bacteria have been given attention. A selected Lactiplantibacillus strain TBRC 20328, with probiotic properties isolated from fermented Isan-style pork sausage (Mam), was evaluated for its safety through whole-genome sequencing and analysis using integrative bioinformatics tools. The metabolic genes were assessed through comparative genome analysis among Lactiplantibacillus species. The genome of the strain TBRC 20328 consisted of one circular chromosome (3.49 Mb) and five plasmids (totaling 0.25 Mb), encoding 3056 and 284 protein-coding genes, respectively. It exhibited an average nucleotide identity (ANI) with other Lactiplantibacillus pentosus strains of over 95%. Whole-genome analysis confirmed the absence of virulence and antimicrobial resistance genes, supporting its safety for food applications. Functional annotation revealed clusters for bacteriocins (plantaricin EF and pediocin) and polyketides, indicating potential roles in biopreservation and host interactions. Genes involved in the biosynthesis of some short-chain fatty acids and exopolysaccharides were also identified. Comparative genomic analysis across 33 other Lactiplantibacillus strains identified 2380 orthogroups, with 94 unique to the Lp. pentosus group. These included gene clusters involved in malonate decarboxylation, leucine biosynthesis, and 5-oxoprolinase activity. Such distinct genomic features emphasize the sustainable biotechnological potential and safety of Lp. pentosus TBRC 23028. Together, the findings highlight its promise as a safe and functional probiotic candidate with broad applications in functional food development and precision fermentation technologies.