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Unveiling the Probiotic Properties of Lacticaseibacillus paracasei UFTM 2.9 Through Probiogenomic Analysis.

Lactic acid bacteria (LAB) comprise a group of Gram-positive bacteria with biotechnological applications. LAB, including Lacticaseibacillus spp., are recognized as potential probiotics due to their ability to confer benefits to the host. Here we employ probiogenomic and in vitro analyses to characterize the probiotic potential of Lc. paracasei UFTM 2.9, a LAB that previously demonstrated probiotic properties in vitro. The draft genome of Lc. paracasei UFTM 2.9 comprises 127 contigs, totaling 3 216 252 base pairs, with a GC content of 46.20%. The bacteria showed metabolic versatility, growing in five carbon sources. A total of 170 genes potentially associated with probiotic characteristics were identified, with functions linked to stress resistance (n = 106), adhesion (n = 12), biosynthesis of vitamins (n = 10), and others. No virulence genes or CRISPR elements were detected, and two phages were identified in Lc. paracasei UFTM 2.9. Gene clusters encoding bacteriocins were detected and confirmed in vitro. Lc. paracasei UFTM 2.9 inhibited all indicator bacteria tested (n = 12), including strains of Listeria innocua, Staphylococcus aureus, Streptococcus agalactiae, and Escherichia coli. The results indicate the potential use of Lc. paracasei UFTM 2.9 as a probiotic, considering its genetic potential to express traits of interest and survive in the gastrointestinal tract (GIT).

Probiotics

Screening of Fermentative Strains for Reducing the Allergenicity of a Whey Protein-Soy Protein System and Genomic Characterization of the Selected Strain.

Dual-protein systems combining whey protein isolate (WPI) and soy protein isolate (SPI) offer complementary nutritional benefits but are limited by the presence of major allergens. Lactic acid bacteria (LAB) fermentation provides a promising strategy to mitigate this limitation. In this study, Lacticaseibacillus paracasei JM053, selected from 13 LAB strains based on phenotypic screening, significantly reduced the in vitro allergenicity of the dual-protein system, increasing the IgE-binding inhibition rate to 48.75%. Whole-genome sequencing and characterization of JM053 revealed a comprehensive proteolytic system, including the proline-specific peptidase genes pepX and pepQ, which may contribute to the degradation of allergenic peptide sequences. Combined with in silico bioinformatic analysis, potential cleavage sites within the linear epitopes of the dual-protein system were predicted based on the substrate specificity of the identified proteases, offering a testable hypothesis for the strain's mechanism of action. In addition, in vitro safety assessment and genomic analysis supported the safety potential, stress tolerance, and probiotic characteristics of JM053. Collectively, this study provides a valuable candidate strain for the development of hypoallergenic dual-protein products and offers preliminary genomic insights into LAB-mediated allergenicity reduction.

Lacticaseibacillus paracasei

Hybrid genome assembly and phenotypic assays reveal carbohydrate metabolism diversity in Lacticaseibacillus strains.

Investigation of carbohydrate metabolism in lactic acid bacteria is essential for the rational selection of strains for fermentation processes, particularly in emerging applications involving non-conventional substrates or building of synthetic microbial consortia. However, establishing robust genotype-phenotype relationships remains challenging, as gene presence alone often fails to explain observed metabolic traits without considering the genomic context and regulatory architecture. In the present study, we combined hybrid genome assembly (Illumina and Oxford Nanopore) with high-throughput phenotype profiling (Biolog GENIII and PM2A) to investigate carbohydrate utilization in five Lacticaseibacillus strains. Phenotypic assays revealed clear intra- and inter-specific variability in substrate utilization. We therefore investigated whether such differences could be attributed to the organization and regulatory context of carbohydrate-associated loci, rather than to gene presence alone. Functional annotation based on COG and CAZyme databases revealed candidate genomic regions potentially involved in carbohydrate metabolism. Comparative analysis between predicted and experimentally observed substrate usage highlighted specific loci associated with carbohydrate utilization profile. The trehalose (tre) operon was conserved across all strains, while at least two distinct cellobiose-associated loci were detected in each genome. Despite the presence of these loci, L. paracasei strains were unable to metabolize cellobiose, a phenotype likely linked to the presence of a downstream TetR-type transcriptional repressor within the cellobiose (cel) operon. Additionally, a genomic region uniquely found in L. rhamnosus strains was associated with gentiobiose utilization, consistent with phenotypic observations. Overall, these findings highlight the importance of integrating phenotypic validation with complete genome context to support the identification of candidate structural and regulatory determinants of carbohydrate utilization in lactic acid bacteria. KEY POINTS: • Phenotype microarrays reveal metabolic traits of interest in isolated strains. • Regulatory context is key to understanding carbohydrate metabolism differences. • Basis of subspecies-dependent cellobiose metabolism in L. paracasei is provided.

Carbohydrate Metabolism