{"paper_id":"30912daa-1501-45c7-bff0-759ee2d95ca9","body_text":"Genomic Characterization and Probiotic Properties of Lactiplantibacillus pentosus Isolated from Fermented Rice | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Genomic Characterization and Probiotic Properties of Lactiplantibacillus pentosus Isolated from Fermented Rice Athira Cheruvari, Rajagopal Kammara This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4845642/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 21 Oct, 2024 Read the published version in Probiotics and Antimicrobial Proteins → Version 1 posted 13 You are reading this latest preprint version Abstract This study focused on a potential probiotic strain of Lactiplantibacillus pentosus (strain krglsrbmofpi2) isolated from traditional fermented rice in India. The study aimed to conduct preliminary genetic and phenotypic characterization. The strain has a genome size of 3.7 Mb, a GC content of 46%, and 3192 protein-coding sequences, as determined by genomic analysis. Subsequently, various bacteriocins, the CRISPR Cas system, phage genes, plasmids, pathogenicity, and antibiotic resistance were identified using bioinformatic methodologies. Biochemical and biophysical examinations improved our understanding of hydrophobicity, antioxidant activity, antibiotic resistance, auto-aggregation, co-aggregation properties, and tolerance to the simulated gastrointestinal condition. The safety assessment of the isolated L. pentosus was evaluated based on the hemolytic activity. The strain has demonstrated strong antibiotic activity against pathogens considered WHO priorities, such as Salmonella enterica subsp. enterica ser. Typhi, Clostridium perfringens , Escherichia coli , Listeria monocytogenes , Staphylococcus aureus , and Vibrio cholerae . Our findings suggest that gaining a deep understanding of the genetic and functional characteristics of the L. pentosus strain could pave the way for its application as a beneficial probiotic in the food industry. L. pentosus whole genome sequence bacteriocin gastrointestinal tract antimicrobial Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Introduction Many ethnic cultures consider fermented foods essential to their customs and traditions [ 1 ]. Fermentation has been used for centuries to extend the shelf life of food and improve its flavour, texture, and nutritional value [ 2 ]. During fermentation, an organism converts a carbohydrate into an acid or alcohol. The most common yeast strain used in fermentation processes is Saccharomyces cerevisiae , which is used to produce wines, beers, and ciders [ 3 ]. Lactic acid bacteria (LAB) are also associated with fermented foods, including Enterococcus , Lactobacillus , Lactococcus , Leuconostoc , Pediococcus , and Weissella species. Additionally, cheese-derived Bifidobacterium and species of Bacillus in fermented legume-based foods are included in this category [ 4 , 5 ]. These microorganisms, consumed through food, play a crucial role in maintaining good health and are called pro-biotics [ 6 ]. Probiotics have been used to modify the intestinal microbiota to improve the health of both humans and animals. To reduce the risk of gastrointestinal (GI) infections, several well-characterized lactobacilli and bifidobacterial strains are already available for human consumption [ 7 ]. The consumption of probiotics has been shown to improve intestinal health by regulating the microbiota, stimulating the development of the immune system, improving nutrient bioavailability, reducing the risk of various cancers, enhancing lactose digestion, preventing cardiovascular diseases, and lowering the risk of certain other diseases [ 8 , 9 ]. Probiotics are generally considered safe, although some LAB species have been linked to opportunistic infections. Further research is needed to determine the advantages and adverse effects of their use [ 10 , 11 ]. Therefore, when evaluating the general safety of probiotic-based foods and beverages, factors such as the source and nature of the probiotics, their pathogenicity, the method of administration, their potential to carry antibiotic resistance genes, the level of exposure, the host’s health, and their intended use must be taken into account [ 12 ]. A microorganism’s potential to serve as a probiotic depends on its pathogenicity, non-poisonous nature, allergenicity, carcinogenicity, or mutagenicity, and it should not have any adverse effects. It should be compatible with the food matrix and associated handling and storage requirements. It should be able to colonize the intestinal mucosa and survive through the gastrointestinal tract (adhesion characteristics, tolerance to acid and bile). Additionally, the host should benefit from it [ 13 ]. Furthermore, the strain must endure at the administration sites, multiply, and colonize for maximal activity in this particular spot. Moreover, the immune system should be able to tolerate it [ 7 ]. India offers various region-specific fermented food products and beverages [ 14 ]. These traditional foods are typically prepared at home through natural fermentation. Despite their long history, many of these fermented foods have not been thoroughly studied from a scientific perspective. Therefore, utilizing modern food microbiology, such as sequence-based taxonomy, and examining the bioactive compounds present in these foods through metabolomics and other technologies, could help validate the value of this traditional food culture [ 15 ]. The Indian Himalayan Region is known for its traditional fermented foods and alcoholic beverages, making it a gastronomic hotspot. Alcoholic drinks have deep ceremonial significance for the ethnic communities of this region and have been consumed at cultural, social, and religious events for generations [ 16 ]. The main ingredients used in these foods include cereals, vegetables, meat, unripe fruits, legumes, and bamboo shoots [ 17 ]. Ethnic fermented foods, such as bhaturu, marchu, dosha, chilra, bedvin roti, seera, pinni, pakk, thuktal, sepubari, bari, churpa, and aska, are frequently made in Himachal Pradesh. Cereals are the main ingredient in these products. Himachal Pradesh is home to several traditional fermented alcoholic drinks made from millets, apples, wild apricots, and grapes, including sura, lugri, kinnauri, chakti, chulli, behmi, and ara [ 16 ]. L. pentosus strains are lactic acid bacteria (LAB) commonly found in various fermented olives from the Mediterranean region, with several strains exhibiting probiotic characteristics [ 18 ]. Due to its genetic diversity and adaptability, it possesses a range of molecular defenses to withstand harsh environmental conditions during processing and consumption [ 19 ]. Research indicates that L. pentosus derived from vegetable fermentation offers numerous health benefits, including cholesterol reduction, antioxidant, anticancer, immunomodulatory activities, and antagonistic and antibacterial effects against gut pathogens [ 20 ]. The safety and probiotic properties of L. pentosus , a GABA-producing strain isolated from Thai pickles, were evaluated through whole genome sequencing, demonstrating its safety and absence of transferable antimicrobial resistance (AMR) genes. The genome does not contain genes associated with biogenic amines, antimicrobial compounds, or clinically relevant toxins [ 21 ]. Probiotic traits and health advantages vary according to the strain. Every strain is distinct. The primary goal of this study was to report on isolating probiotic L. pentosus (strain krglsrbmofpi2) from fermented foods from unknown ethnic groups in Himachal Pradesh, India. Its genomic and probiotic properties were assessed to determine whether the isolated strain could be used as a safe food preservative or as a probiotic food. Materials and Methods Isolation of Probiotic Bacteria Various fermented foods, including fermented rice sourced from villages in Himachal Pradesh, were collected for investigation. From the collected samples, a 1.0% (1.0 mL) aliquot was mixed with 9 mL of peptone broth and incubated at 37°C under aerobic conditions (200 RPM agitation in New Brunswick Innova 44, incubator shaker, Ham-burg, Germany.) for a duration of 24 h. Subsequently, the sample was spread-plated onto de Man, Rogosa, and Sharpe (MRS) agar plates to facilitate the isolation of various bacterial strains. Salmonella enterica subsp. enterica ser. Typhi ( S. Typhi) was used to evaluate the antibacterial activity of the isolated strains. The selected strain, active against S. Typhi, was sub-cultured (1.0%) in MRS broth and incubated at 37°C for 24 h, 200 RPM, in a shaker incubator. The nature of the antimicrobial compound was determined by testing its activity in the presence of different enzymes. The enzyme solutions (1 mg/mL) were prepared as follows: trypsin (in Tris-HCl 40 mM, pH 8.2), pepsin (in 0.002 M HCl, pH 2.0), chymotrypsin (in 50 mm Tris-HCl, pH 8.0), catalase (in 20 mM HCl, pH 2.0), protease (in water), lysozyme (in 10 mM Tris-HCl, pH 8.0), amylase (in water), lipase (in water). All enzymes were from Sigma-Aldrich (St. Louis, MO, USA). Then, 1.0 mL of cell-free supernatant (CFS) solution was added to 1.0 mL of the enzyme solutions. After 1 h of incubation at 37°C, samples were boiled for 5 min at 100°C to inactivate the enzyme. Concentrated CFS (60 µL, Enzyme-treated CFS concentrated using a vacuum concentrator, Eppendorf, Hamburg, Germany) was used to estimate their antimicrobial activity through an agar-well diffusion assay against S. Typhi [ 22 – 24 ]. Preliminary Characterization of Isolates Gram staining and scanning electron microscopy analyses are standard techniques initially used to characterize isolated bacteria [ 25 ]. Next, a variety of biochemical tests were performed, such as the catalase test (using S. Typhi as a positive control), sugar utilization tests, growth in various media, growth at different pH (2.0 to 10), growth at multiple concentrations of NaCl (0 to 10%) [ 24 , 26 ] and varying concentrations of phenol (0.1–0.4%) [ 27 ]. This growth investigation used 1.0% overnight-grown cells and incubated for 24 h at 37°C, 200 RPM. Culture grown in MRS broth was used as a control. The optical density (OD) was measured at 600 nm after 24 h [ 27 , 28 ]. The following different media were used in the study: M17 Broth, Sabouraud Dextrose Broth, Hi-veg (SD), Brain Heart Infusion Broth (BHI), Nutrient Broth (NB), Luria Bertani Broth (LB), Yeast Extract Peptone-Dextrose Broth (YPD), Elliker Broth (ELK), Bifidobacterium broth (BB), Tryptone soya broth (SB), de Man Rogosa, and Sharpe Broth (MRS) (all media from Hi Media) [ 29 ]. Moreover, LAB was identified at the species level using additional biochemical tests like sugar utilization pattern and other properties by using the HiLacto identification kit (KB020, [ 30 ], Hi Carbo kit (KB009A/KB009B1) [ 31 ], Hi Bacillus Identification Kit (KB013). All kits are from HiMedia, Mumbai, India [ 32 ]. DNA Extraction Thermo Fisher Scientific’s Gene JET Bacterial Genomic DNA Purification Kit was used to extract genomic DNA. The kit’s instructions were meticulously followed. A nano-drop spectrophotometer was used to verify the amount and quality of genomic DNA (Thermo Fisher Scientific, Waltham, MA, USA) [ 33 ]. Phylogenetic Analysis The identity of cultures was verified by sequencing 16S rRNA using bacterial universal primers 27F (5′-AGAGTTTGATCCTGGCTCAG-3′) and 1492R (5′-GGTTACCTTGTT ACGACTT-3′) [ 34 ]. To identify the nearest neighbour in the evolutionary tree, the 16S rRNA sequences of similar organisms were taken from the NCBI database (National Centre for Biotechnology Information) and compared with the sequences of isolated isolates. The Tamura-Nei model and the Maximum Likelihood technique were used to estimate the evolutionary history [ 35 ]. Comparative analyses of 10 nucleotide sequences enabled the identification of the closest relatives in the phylogenetic tree constructed using the Neighbour-joining (NJ) method with a p-distance model in MEGA X software (Molecular Evolutionary Genetics Analysis, version 10.0.0, https://megasoftware.net , accessed on 21 March 2019) [ 36 ]. Genomic Characterization Genome Sequencing and Mapping Whole genome sequencing of the isolate was performed by Sandor Speciality Di-agnostics (Hyderabad, India) using Illumina Hi Seq 1000 technology [ 33 ]. The results were submitted to the National Centre for Biotechnology Information (NCBI). Contigs containing genomic DNA were built using the SPAdes v. 3.13.1 assembler. The NCBI PGAP (Prokaryotic Genome Annotation Pipeline) contributed to the annotation [ 37 ]. With the help of the CG View server, a circular genome map was created ( https://cgview.ca/ , accessed on 14 December 2022) [ 38 ]. Bioinformatic Analysis Different bioinformatics tools were used to annotate and identify functional genes. Probiotic Factors Probiotic genes were identified using blast analysis by searching the entire genome sequence for sequence similarity [ 25 ]. The functional annotations of the genome were manually examined for the presence of the essential probiotic genes through prokka-generated annotation based on prior studies [ 39 ]. Various proteins involved in stress response, resistance, adhesion, aggregation, and immunomodulation were examined [ 40 ]. Bacteriocin and Secondary Metabolite BAGEL v.4.0, an online web server, was used to mine the antimicrobial peptides (bacteriocin) produced by bacteria. The assembled genome in FASTA format was uploaded and processed using the default settings in BAGEL v.4.0 ( http://bagel4.molgenrug.nl , accessed on 14 December 2022) [ 41 , 42 ]. Secondary metabolite production was analyzed using the bacterial versions of antiSMASH 6.0 (accessed on 14 December 2022), one of the most popular software programs for this task. It is based on profile hidden Markov models of genes typically found in specific Gene clusters [ 43 , 44 ]. Plasmids The genome of the isolated strain was examined for the presence of plasmids using the Plasmid Finder web tool version 2.0.1 ( https://cge.food.dtu.dk/services/PlasmidFinder/ , accessed on 14 December 2022). A minimum coverage of 60% and a minimum identity percentage of 90% were added to the parameters. Plasmid Finder detects replicons in WGS and assigns the plasmid under study to incompatibility (Inc.) groups, referring to the plasmid’s GenBank accession number as the reference for that group [ 45 – 47 ]. CRISPR/CRISPR-Cas The CRISPR regions were detected with a CRISPR online detection tool, CRISPR Cas finder ( https://crisprcas.i2bc.paris-saclay.fr/CrisprCasFinder/ , accessed on 14 December 2022). CRISPR refers to a specific family of tandem repeats in various bacterial genomes. CRISPR-Cas9 is a prokaryote adaptive immune system that protects cells from DNA virus infections [ 48 ]. Prophage Regions The PHAge Search Tool Enhanced Release (PHASTER) web server ( www.phaster.ca , accessed on 14 December 2022) identified potential prophage locations. The PHASTER finds, annotates, and visually shows prophage sequences found in bacterial genomes or plasmids. Bacteriophage genes, both functional and non-functional, may be found in more than 20% of bacterial genomes. Prophage sequences can explain a significant portion of the variance found among bacterial species or clades. Prophage sequences may also aid in developing pathogenicity, antibiotic resistance, exploration of novel ecological niches, and strengthened adhesion in certain bacteria [ 49 – 51 ]. Pathogenicity Human pathogenicity and virulence were predicted by the Pathogen Finder v.1.1 website (accessed on 15 December 2022, from https://cge.cbs.dtu.dk/services/PathogenFinder/ ). It is a web server that predicts the pathogenicity of bacteria by analyzing the proteome, genome, or raw data given by the user [ 52 ]. Antibiotic Resistance Genes Antibiotic resistance genes were found using the Comprehensive Antibiotic Research Database (CARD Database, https://card.mcmaster.ca/analyze , accessed on 16 December 2022). It contains bioinformatic tools for identifying antibiotic resistance genes in whole genome or partial genome sequence data, including unannotated raw sequence assembly contigs. It is a robustly curated database in a user-friendly format that assembles over 1600 known antibiotic resistance genes, allowing sophisticated antibiotic resistance analysis and query in a way that will benefit the broader biomedical research community [ 53 – 55 ]. Probiotic Properties Acid and Bile Tolerance Following a 24 h incubation period in MRS broth, the isolated strain was sub-cultured at a concentration of 10% in three different growth media: MRS broth adjusted to pH 3.0 to assess acid tolerance, MRS broth supplemented with 0.15% bile salt to evaluate bile tolerance, and standard MRS broth serving as the control (initial OD—0.5). The adjustment of pH to 3.0 was achieved using 1.0 N HCl. Cultures were maintained aerobically (200 RPM agitation) at 37°C throughout the experiment [ 56 – 58 ]. Samples were withdrawn at intervals of 0, 3, 6, 9, 12, and 24 h for evaluation. Growth dynamics were assessed by measuring optical density at 600 nm (OD600) using a spectrophotometer (DLAB SP-V1000, Los Angeles, CA, USA). Additionally, serial dilutions of the cultures were plated onto MRS agar plates, followed by incubation for 24 h at 37°C. Colony-forming units per millilitre (CFU/mL) were enumerated to quantify bacterial growth. All counts were obtained in duplicate and expressed as mean ± SD [ 59 ]. Finally, the acid and bile tolerance/percentage of survivability was estimated by the following equation: Bile/Acid Tolerance (%) = [OD600 of test/OD600 of control)] × 100 [ 60 – 62 ]. Bile Salt Hydrolase Assay Bile salt hydrolase (BSH) plays a crucial role in the metabolism of bile acids, particularly in the intestine. This enzyme facilitates the hydrolysis of bile acids conjugated with glycine and taurine, liberating free bile acids and amino acid residues. BSH activity of intestinal bacteria is one of the indirect ways of decreasing the cholesterol level in the human body [ 63 ]. The BSH activity of the isolated strain was detected using a plate as-say, with slight modifications. Using a sterile cork borer (6.0 mm diameter), wells were made on a bile salt agar (HiMedia, Mumbai, India) plate, and the fresh overnight cultures (100 µL) were added. Plates were incubated in an aerobic environment for 24 h at 37°C. MRS agar plates without bile salt were used as a control. The presence of precipitated bile acid (cholic acid) around colonies (opaque halo) or the formation of opaque granular white colonies was considered a positive reaction. Each sample was analyzed twice [ 64 – 66 ]. Lysozyme Tolerance Lysozyme tolerance was assessed with a 24 h growth culture, washed with phosphate buffer saline (PBS) solution (NaCl—1.37 M, KCl—27 mM, Na2HPO4—10 mM, KH2PO4—18 mM, pH 7.0), resuspended the 1.0 mL of collected cells in 10 mL of PBS solution containing 0.01% lysozyme (Sigma-Aldrich, St. Louis, MO, USA), and incubated at 37°C for 1 h. Subsequently, cell viability was estimated as CFU/mL by serially diluted cells plated on an MRS agar plate. All counts were obtained in duplicate, and mean values were taken [ 27 , 67 ]. Gastrointestinal Tolerance Isolated strains were cultured for 24 h in MRS broth at 37°C, and the absorbance at 600 nm (OD600) was measured using a spectrophotometer after incubation. Cells were pelletized by centrifugation (6000 RPM for 10 min at room temperature) and re-suspended in sterile MRS broth. Culture at 10% (0.5 OD) was added to MRS broth containing stimulated gastric fluid (0.3% pepsin at pH 3), intestinal fluid (0.1% pancreatin and 0.15% bovine bile salt at pH 8), and culture in MRS broth was used as control [ 57 ]. These cell suspensions were incubated for 24 h at 37°C, 200 RPM. During the incubation, absorbance (OD at 600 nm) and the viable cell counts (CFU/mL) were determined for 0, 3, 6, 9, 12, and 24 h. The experiments were repeated twice, and the mean values were taken [ 27 , 68 ]. The following equation measured the percentage of bacterial survival: Percentage of Survivability/Tolerance = (Growth of Test/Growth of Control) × 100 [ 69 ]. Cell Surface Hydrophobicity Cell surface hydrophobicity refers to the tendency of microbial cell surfaces to interact with hydrophobic molecules or surfaces. It plays a significant role in microbial processes, including adhesion to host tissues, biofilm formation, and interactions with environmental surfaces. The measurement of cell surface hydrophobicity is crucial in understanding the behaviour and physiology of microorganisms. The overnight culture of the isolated strain was pelletized by centrifugation (6000 RPM, 10 min), washed twice using sterile phosphate buffer saline (PBS), and re-suspended in 3.0 mL of the same buffer. The initial absorbance (A0) was adjusted to 0.5 OD at 600nm. Then, 1.0 mL of xylene (Merck, Darmstadt, Germany)/chloroform/hexadecane (HiMedia, Mumbai, India) was added to the bacterial suspension and vortexed for 2 min [ 63 , 70 ]. The mixture was subjected to phase separation by incubating at 37°C for 1 h, and the absorbance of the aqueous phase (A1) was measured using a spectrophotometer at 600 nm. The experiments were repeated twice, and the mean values were taken [ 28 ]. The following equation measured the percentage of hydrophobicity: % Cell surface hydrophobicity = (1 − A1/A0) × 100. DPPH (2,2-Diphenyl-1-picrylhydrazyl) Free Radical Scavenging Assay The DPPH assay was used to measure the capacity to scavenge free radicals. The overnight culture was centrifuged for 15 min at room temperature at 6000 RPM, and the cell-free supernatant (CFS) was collected in a separate tube. CFS (2.0 mL) was mixed with 2,2-diphenyl-1-picrylhydrazyl (2.0 mL, DPPH—6 mg/100 mL in methanol). Methanol served as the blank, while 2.0 mL of DPPH in 2.0 mL of methanol served as the control. The reaction mixture was thoroughly vortexed and allowed to sit in the dark at 37˚C for 30 min. Absorbance was recorded at 517 nm [ 71 ]. Ascorbic acid (10 µg/mL, Sigma-Aldrich, St. Louis, MO, USA) was used as a positive control in distilled water. Each sample analysis was conducted in duplicate to ensure accuracy and reliability, and the following formula was applied to determine the antioxidant activity: Free radical scavenging activity (%) = (Ac − As)/Ac × 100 As is the sample absorbance, and Ac is the control absorbance at 517 nm [ 60 ]. Proteolytic Activity Assays The isolated strain was screened for proteolytic activity by an agar-well diffusion method on skim milk containing 2.0% (w/v) agar (Hi-Media). Actively growing culture (60 µL) and CFS (60 µL) were added to the wells made in an agar plate (skim milk agar) and incubated at 37°C. Trypsin (1.0 mg/mL) was used as a positive control. A clear zone around the agar-well-containing bacteria indicates a positive result. This methodology enabled the qualitative assessment of the strain’s ability to hydrolyse proteins, as evidenced by the formation of clear zones resulting from casein degradation within the skim milk agar medium [ 72 ]. Antibiotic Susceptibility Testing The antibiotic susceptibilities of isolated bacteria were determined by the disc diffusion method, as recommended by CLSI (Clinical and Laboratory Standards Institute, Wayne, PA, USA). Thus, 100 µL of 0.5 OD culture mixed with MRS agar was plated. Antibiotic discs Universal-1 OD308 and Dodeca Universal-XII DE027 (HiMedia, Mumbai, India) were placed on the agar plates with culture. They were incubated at 37°C for 24 h. The diameter of zones was measured, and the isolates were identified as sensitive, intermediate, or resistant according to CLSI tables and guidelines. The experiments were repeated twice [ 73 ]. Hemolytic Activity The isolated bacteria’s hemolytic activity was determined by adding the 24 h grown culture and cell-free supernatant to the wells on blood agar plates containing 5.0% (w/v) sheep blood and incubating at 37°C for 24 h. After incubation, the plates were examined for β-hemolysis (Clear zone of inhibition), α-hemolysis (Greenish zone), and non-hemolytic activities (γ-hemolysis). Triton X 100 (HiMedia, Mumbai, India) and Nisin (1 mg/mL, HiMedia, Mumbai, India) were used as controls. Each sample was examined twice. [ 60 ]. Auto-Aggregation and Co-Aggregation The ability of isolates to auto-aggregate was evaluated by 24 h grown culture. OD at 600 nm was adjusted to 0.5, and cells were centrifuged (6000 RPM, 10 min), and pellets were resuspended in a standard saline solution. The absorbance was measured at 0 h and 3 h by a spectrophotometer at 600 nm without shaking the cell suspension [ 74 ]. The au-to-aggregation was estimated as follows: Auto-aggregation (%) = (1 − At/A0) × 100, A0 is the initial absorbance, and At is the absorbance at 3 h [ 75 ]. The isolate’s co-aggregation with pathogenic bacteria was evaluated by mixing pathogenic bacteria (3.0 mL) and probiotic cells (3.0 mL) in a sterile tube. Both cells had 0.5 OD (at 600 nm). We mixed it well and incubated it at 37°C in static conditions. The absorbance was measured at OD 600 nm after 3 h. Pathogenic and probiotic bacteria were cultured separately, and their absorbance was taken as control. The percentage of co-aggregation was calculated according to the formula: Co-aggregation (%) = [(Ax + Ay)/2 − Axy]/ [(Ax + Ay)/2] × 100, where Ax—represents the absorbance of the probiotic strain, Ay—represents the absorbance of the pathogenic bacteria under study, Axy—represents the absorbance of the mixture of both [ 76 ]. The food pathogen S. Typhi was the pathogen used for the co-aggregation assay. Co-aggregated cells and auto-aggregated cells after 3 h of incubation were visualized under microscopy using Gram staining [ 77 ]. Antibacterial Activity The antibacterial activity of the LAB strains against food pathogens was determined using the agar well diffusion assay. The pathogenic indicator bacteria used for the study included Salmonella enterica subsp. enterica ser. Typhi (new isolate), Salmonella enterica subsp. enterica ser. Paratyphi (MTCC 735), Listeria monocytogenes (MTCC 839), Streptococcus mutants (MTCC 497), Streptococcus thermophilus (New isolate), Staphylococcus aureus (MTCC 1430), Pseudomonas aeruginosa (MTCC 1934), Pseudomonas putida (MTCC 2492), Vibrio cholerae (MTCC 3904), Vibrio harveyi (MTCC 7954), Corynebacterium callunae (MTCC 700), Enterococcus gallinarum (MTCC 7049), Clostridium perfringens (MTCC 450), Escherichia coli (k2 strain) and Bacillus cereus (MTCC 430) [ 78 ]. For the well-diffusion assay, 100 µL of an overnight culture of the indicator strain was mixed with BHI soft agar (0.75% agar in BHI Broth) and poured into a BHI agar plate. A 6.0 mm diameter well was made in the agar, and 60 µL (1.0 mg/mL protein concentration of protein present in CFS) of 10-fold concentrated cell-free supernatant was added, and incubated the plates for 12 h to measure the zone of inhibition. The experiments were repeated twice [ 79 ]. Statistical Analysis The statistical data analysis was conducted using GraphPad Prism version 8.0.2 for Windows. All data were expressed as mean ± standard deviation. One-way analysis of variance (ANOVA) was employed to compare a single parameter across multiple groups. A significance threshold of p < 0.05 was utilized, where statistical significance was determined. Results and Discussion Preliminary Characterization of LAB Isolates The strains isolated from fermented rice underwent initial characterization through physiological and biochemical testing. A diverse array of both Gram-positive and Gram-negative strains were obtained. Subsequent antibacterial assays against S. Typhi revealed that only one strain exhibited significant activity, as evidenced by the observed zone of inhibition (Fig. 1 A). The antimicrobial activity present in the cell-free supernatant (CFS) was subjected to enzymatic treatment with various enzymes, including trypsin, chymotrypsin, pro-tease, catalase, lysozyme, amylase, and lipase. Upon treatment, a decrease in antimicrobial activity was observed in the presence of protease, chymotrypsin, and trypsin, suggesting a proteinaceous nature of the antimicrobial compound. The zone of inhibition was measured at 16 mm for protease, 15 mm for chymotrypsin, and 18 mm for trypsin, while all other enzymes resulted in a consistent zone of inhibition of 19 mm. These findings underscore the susceptibility of the antimicrobial activity to enzymatic degradation by proteolytic enzymes, indicating its dependence on proteinaceous components for antimicrobial efficacy. Such insights are crucial for elucidating the biochemical nature of the antimicrobial compounds present in the CFS. Morphological examination indicated that colonies of this active strain were circular, smooth, and elevated (Fig. 1 B), with Gram staining confirming this Gram-positive nature (Fig. 1 C). Scanning electron microscopy (SEM) analysis further corroborated the rod-shaped morphology of the strain (Fig. 1 D). Further characterization included catalase testing, which indicated that the isolated strain was catalase-negative, contrasting with the catalase-positive S. Typhi used as a control. Growth assessment under varied conditions encompassed different culture media, pH levels, salt concentrations (NaCl), and phenol concentrations. Among the media tested, MRS broth supported the most rapid growth of the isolated strain, with significantly lower growth observed in other media (Fig. 2 A). Optimal growth occurred at pH 7.0 at 37°C (Fig. 2 B), while exposure to increasing NaCl concentrations resulted in a progressive reduction in growth, with a notable decrease observed from 6.0% NaCl (Fig. 2 C). The observed characteristics’ parallel findings reported in L. pentosus strains were isolated from naturally fermented Aloreña green table olives [ 80 ]. Evaluation of the strain’s tolerance to phenol revealed its ability to survive up to a 0.4% phenol concentration; however, it had diminished growth compared to control conditions, particularly at higher phenol concentrations (the growth decreased with increasing phenol concentration from 0.1 to 0.4% compared to the control) (Fig. 2 D). These findings underscore the adaptability and resilience of the isolated strain to various environmental conditions. The biochemical analysis indicated that the isolated bacteria could utilize all tested sugars, as outlined in Supplementary Table S1 . It includes lactose, sucrose, fructose, glucose, sorbitol, mannitol, etc. Consistently, L. pentosus TEZU174 also exhibited a comparable sugar utilization profile [ 81 ]. Additionally, the results of various biochemical reactions, such as Voges-Proskauer’s test, citrate utilization, ONPG, and nitrate reduction, are detailed in Table 1 . Table 1 Biochemical test results of the L. pentosus strain. Sl No. Test Principle Results 1. Malonate Malonate utilization −ve 2. Voges Proskauer’s Detects acetoin production −ve 3. Citrate Citrate utilisation −ve 4. ONPG Detects Beta galactosidase +ve 5. Nitrate Reduction Detects Nitrate reduction −ve 6. Catalase Detects Catalase activity −ve 7. Arginine Arginine utilisation −ve 8. Esculin Hydrolysis Detects Esculin Hydrolysis activity +ve +ve: indicates positive results; −ve: negative results. Confirmation of its classification as a Lactiplantibacillus species was attained through 16S rRNA gene sequencing. Phylogenetic analysis unveiled a striking 99.6% similarity with strains, such as Lactiplantibacillus pentosus LMEM (MK240372.1), Lactiplantibacillus pentosus GCHI (MK245998.1), and several Lactiplantibacillus plantarum strains, including Lactiplantibacillus plantarum SCHI (MK246005.1), and Lactiplantibacillus plantarum MA8-6 (MG755354.1) (refer to Table 2 and Fig. 3 ). Table 2 Microbial identification using 16S rRNA-based molecular method. Sequences of strains having significant alignments are given in the table. Sl No. Description Identity Accession No. 1. Lactiplantibacillus plantarum strain SCHI 99.63% MK246005.1 2. Lactiplantibacillus pentosus strain GCHI 99.63% MK245998.1 3. Lactiplantibacillus pentosus strain LMEM 99.63% MK240372.1 4. Lactiplantibacillus plantarum strain MA8-6 99.63% MG755354.1 5. Lactiplantibacillus plantarum strain NWAFU1580 99.63% MK045823.1 6. Lactiplantibacillus plantarum strain TTF18 99.63% MK028367.1 7. Lactiplantibacillus plantarum strain TTF17 99.63% MK028366.1 8. Lactiplantibacillus plantarum strain TTF16 99.63% MK028365.1 9. Lactiplantibacillus plantarum strain TTF15 99.63% MK028364.1 10. Lactiplantibacillus plantarum strain TTF14 99.63% MK028363.1 Further confirmation of its identity as Lactiplantibacillus pentosus (formerly known as Lactobacillus pentosus ) strains was achieved through whole genome sequencing. Both datasets, including 16S rRNA gene accession (MN165450.1) and WGS GenBank assembly accession (GCA_009295675.1), were duly submitted to NCBI. Genomic Characterization The genomic analysis of Lactiplantibacillus pentosus reveals a composite structure consisting of 55 contigs (667,623 base pairs), with a total size of 3.7 Mb and a GC content of 46%. Further, 3342 coding sequences (CDSs) are identified within this genome, of which 3192 encode proteins. Comprehensive annotation further discloses the presence of 76 RNA genes, comprising 65 transfer RNA (tRNA), 7 ribosomal RNA (rRNA), and 4 non-coding RNA (ncRNA) genes. Figure 4 , generated through the CG View Server, provides a visual representation of the genome map, elucidating the arrangement of coding sequences on the strand, contigs, GC content, GC skew, and other pertinent genomic attributes. Additionally, supplementary tools enhance the analysis by identifying various genomic elements and properties. Specifically, Alien Hunter predicts putative Horizontal Gene Transfer (HGT) events, Phigaro detects prophage regions, Mobile OG-db identifies mobile genetic elements (MGEs), VirSorter discerns viral signals within microbial genomic data, CRISPR/Cas Finder detects CRISPR arrays along with their associated Cas proteins, and CARD facilitates the detection of antimicrobial resistance genes. Through this comprehensive approach, a thorough understanding of the genomic landscape of L. pentosus is achieved, enabling insights into its genetic makeup and functional potential Bioinformatic Analysis The probiotic characteristics of Lactiplantibacillus pentosus were meticulously investigated through blast analysis and manual examination of various proteins annotated by Prokka. This comprehensive investigation unveiled a spectrum of proteins implicated in stress response, adhesion, aggregation, resistance, and immunomodulation. Detailed information regarding these proteins and their NCBI accession numbers are provided in Table 3 . Table 3 Represents the details about the proteins present in the isolated L. pentosus strain genome that are involved in stress, adhesion, aggregation, and resistance with NCBI accession number. Sl No. Factors Proteins Accession No. 1. Stress a. Temperature cold-shock protein MPQ17990.1, MPQ19628.1, MPQ20388.1 chaperonin GroEL MPQ20734.1 co-chaperone GroES MPQ20733.1 Hsp20 family protein MPQ17905.1, MPQ18860.1, MPQ19037.1 Hsp33 family molecular chaperone HslO MPQ20807.1 b. pH alkaline shock response membrane anchor protein AmaP MPQ19576.1 c. Other Stress Universal stress protein MPQ18033.1, MPQ18120.1, MPQ18171.1, MPQ18841.1, MPQ19344.1, MPQ19437.1, MPQ20001.1, MPQ20233.1, MPQ20274.1, MPQ20395.1 Peroxide stress protein YaaA MPQ17946.1 GlsB/YeaQ/YmgE family stress response membrane protein MPQ18166.1, MPQ19575.1 Asp23/Gls24 family envelope stress response protein MPQ18230.1, MPQ18256.1, MPQ19578.1, MPQ19579.1 2. Adhesion/Aggregation Adhesion to mucus/epithelial cells/ECM proteins/plasma components/Aggregation Mucus-binding protein MPQ18214.1, MPQ20578.1 Chaperonin GroEL MPQ20734.1 Elongation factor Tu MPQ19535.1 Sortase MPQ19087.1 Glyceraldehyde-3-phosphate dehydrogenase MPQ19912.1 3. Resistance bleomycin resistance protein MPQ17822.1 small multidrug resistance protein MPQ17839.1 copper resistance protein CopZ MPQ18979.1 Moreover, the genome-wide exploration for bacteriocin-encoding genes was conducted using BAGEL 4, revealing the presence of diverse bacteriocin types, including pediocin-like bacteriocin (class IIa), plantaricin E, F (class IIb), and a putative bacteriocin (Bovicin 255 variant). Pediocin showed similarity to bacteriocin from Pediococcus acidilactici. The Plantaricin E and F cluster, a two-peptide bacteriocin with immunity genes, ABC transporter genes, and accessory genes, exhibits similarity to bacteriocin from Lactiplantibacillus plantarum. Additionally, lactococcin is present within the same node. The putative bacteriocin (Bovicin 255 variant) shares similarities with the bacteriocin from Streptococcus mutans UA159. Notably, NCBI protein blast analysis affirmed the existence of garvicin Q family Class II bacteriocin and lactococcin (class IIc), demonstrating the capability of L. pentosus to produce a repertoire of Class II bacteriocins for defensive purposes. Figure 5 A illustrates the outcomes obtained from BAGEL 4. Similarly, other strains of L. pentosus exhibit distinct bacteriocin production profiles. For instance, L. pentosus ZFM94 synthesizes a bacteriocin termed Pentocin, while L. pentosus 124-2 produces two bacteriocins with molecular weights of 26.69 kDa and 17.15 kDa [ 82 , 83 ]. Additionally, antiSMASH software was employed to conduct an in-depth analysis of secondary metabolite production, revealing the presence of two regions harboring secondary metabolites. Notably, region 6 encompasses a RiPP-like cluster (other unspecified ribosomally synthesized and post-translationally modified peptide product) containing a class II bacteriocin region with an ABC transporter and accessory protein, whereas region 32.1 harbors a T3PKS (Type III polyketide synthase) cluster with a hydroxymethylglutaryl-CoA synthase region. A cluster blast analysis of the bacteriocins cluster showed significant similarity to bacteriocin clusters of different L. plantarum strains, with identities ranging from 50 to 58% (Supplementary Figure S1 depicts the results of secondary metabolite search, and Figure S2 depicts the cluster blast results by antiSMASH). This describes the evolutionary relatedness in bacteriocin production. A similar cluster blast was used in L. pentosus strain ZFM94 [ 84 ]. The Plasmid Finder tool identified the presence of a plasmid in the genome with high identity to rep 28 from L. plantarum ( NCBI accession number CP005948). This finding aligns with earlier reports indicating the presence of plasmids in certain L. pentosus strains [ 85 ]. The CRISPR Cas finder detected five sequences with CRISPR genes, indicating an adaptive immunity mechanism against foreign mobile genetic elements [ 86 ]. Nodes 1, 2, 7, 19, and 28 have CRISPR sequences. The genome has CAS-Type IIA and CAS-Type IE. Additionally, prophage regions were predicted by the PHASTER server, with nodes 13 and 15 containing complete and incomplete phage sequences, respectively (Fig. 5 B). These findings are consistent with previous publications, suggesting that the genome of L. pentosus MP-10 included CRISPR Cas genes and prophage regions [ 87 ]. Furthermore, CARD analysis revealed the presence of the Van Y glycopeptide resistance gene cluster by strict hits with 29.63% identity (Fig. 5 C). Loose hits (bit score below 500) showed the presence of 203 antibiotic resistance genes with different resistance mechanisms, including antibiotic efflux, antibiotic target protection, antibiotic inactivation, antibiotic target alteration, reduced permeability to antibiotics, and antibiotic target replacement. Resistance drug classes include glycopeptide antibiotics, peptide antibiotics, macrolide antibiotics, lincosamide antibiotics, cephalosporin, tetracycline antibiotics, aminoglycoside antibiotics, fluoroquinolone antibiotics, disinfecting agents, antiseptics, mupirocin-like antibiotics, rifamycin antibiotics, etc. These findings showed that the strain did not have rigorous antibiotic resistance. These comprehensive analyses collectively enhance our understanding of the probiotic traits, bacteriocin production, genomic architecture, and antibiotic resistance profile of L. pentosus , underscoring its potential for therapeutic applications and biotechnological exploitation. Probiotic Characterization The L. pentosus strain underwent assessment for tolerance to bile and acid, revealing its capacity to withstand 0.15% bile salt concentration and acidic conditions with a pH of 3. The strain’s tolerance dynamics were investigated over time intervals of 0, 3, 6, 9, 12, and 24 h, with survivability percentages calculated from corresponding growth values, as depicted in Fig. 6 A, B. Concurrently, cell viability was evaluated at these time points, and viable counts were recorded, as presented in Supplementary Table S2. Notably, the growth and cell viability levels of the Lactiplantibacillus pentosus strain exhibited an increasing trend over time intervals of 0, 3, 6, 9, 12, and 24 h. However, despite this increase, the percentage of survivability, when compared to the control, demonstrated a decreasing trend. This discrepancy suggests that while the bacterial population grows and maintains viability over time, its ability to survive in the presence of bile salt and acidic conditions decreases relative to the control group. This observation indicates that prolonged exposure to bile salt and acidic environments adversely affects the strain’s survivability, as evidenced by a reduction in its relative resilience compared to the control condition. This observation aligns with prior research conducted by Montoro et al., which explored the survivability of various L. pentosus strains under analogous conditions [ 80 ]. The culture of L. pentosus was screened for bile salt hydrolase (BSH) activity using a direct plate assay. BSH activity is recognized to facilitate bacterial colonization within the gastrointestinal tract [ 88 ]. Notably, the isolated L. pentosus strain produced a discernible white precipitate zone in bile agar media surrounding the colony, indicative of BSH production. Bifidobacterium adolescentis was the positive control used to validate this observation, while S. Typhi was employed as the negative control (Fig. 7 A). Moreover, it is noteworthy that in earlier research, bile salt hydrolase (BSH) activity has been reported in several Lactiplantibacillus species originating from food and human sources. For instance, it has been observed that strains like L. pentosus CHIG have positive BSH activity [ 89 ]. Furthermore, the strain demonstrated 57.32% lysozyme tolerance with a CFU/mL count of 9.5 × 10 7 after 1 h of incubation. This finding underscores the strain’s capability to withstand lysozyme exposure. Gastrointestinal tolerance of the isolated L. pentosus was assessed over intervals of 0, 3, 6, 9, 12, and 24 h. The results were similar to those observed for acid bile tolerance, with survivability percentages of the isolated strain in gastrointestinal conditions compared to control MRS documented in Fig. 6 . Notably, an inverse relationship was observed between survivability percentage and time duration, wherein an increase in incubation time corresponded to a decrease in survivability percentage. The bacterial population is growing and maintaining viability over time; however, its ability to survive in the presence of gastrointestinal conditions decreases relative to the control group. Furthermore, Supplementary Table S2 provides a comprehensive overview of viable counts obtained at each time point, further elucidating the strain’s dynamics in gastrointestinal conditions. These findings underscore the strain’s ability to endure the challenges presented by the gastrointestinal environment. Such insights are pivotal for assessing the strain’s potential as a probiotic agent and its suitability for gastrointestinal health applications. To evaluate its adhesion capability, the strain underwent testing for cell surface hydrophobicity using hydrocarbons, namely xylene, chloroform, and n-hexadecane. The observed hydrophobicity levels were 21.12 ± 0.94% in the presence of xylene, 13.53 ± 0.09% in the presence of n-hexadecane, and 26.72 ± 0.86% in the presence of chloroform. Comparable hydrophobicity profiles have been reported in other strains, such as Lactiplantibacillus pentosus isolated from fermented fish, Lactococcus lactis , and Lactobacillus fermentum [ 90 ]. Probiotic species’ radical scavenging capacity is an important characteristic, indicating their antioxidant nature. This was assessed using the DPPH radical scavenging assay, where the isolated L. pentosus demonstrated 60.70% DPPH activity, showcasing its antioxidative potential. Control ascorbic acid (10 µg/mL) gave 42.75% DPPH activity. Notably, studies by Unban et al. reported even higher DPPH scavenging activity in L. pentosus A14-6 and L. pentosus A26-8 [ 91 ]. A proteolytic assay was conducted using a skim milk agar plate, with trypsin (1.0 mg/mL) as the positive control, resulting in 19 mm of proteolysis. However, the supernatant and pellet of the isolated L. pentosus strain did not exhibit any zone of inhibition in the test, indicating the absence of proteolytic activities. The antibiotic susceptibility profile of the L. pentosus isolate was assessed against a panel of 20 antibiotics, revealing resistance to cefoxitin (CX), amoxicillin/clavulanic acid (AMC), and amikacin (AK). However, sensitivity or intermediate resistance was observed towards all other antibiotics tested (refer to Table 4 and Fig. 7 B, C), indicating the strain is susceptible to most antibiotics, thereby supporting its safety for probiotic applications [ 27 ]. This finding aligns with observations by Cazodo Munoz et al., who noted that out of 59 L. pentosus strains tested, a majority (95%) of strains were resistant to at least 3 antibiotics when evaluated against 15 antibiotics [ 92 ]. Table 4 Represents antibiotic susceptibility results of the isolated L. pentosus strain. Sl No. Antibiotics Concentration (µg/Disc) Zone of Inhibition (in mm) Resistance/ Sensitive Universal-1 OD308 1. Gentamicin (GEN) 10 17 I 2. Amikacin (AK) 30 12 R 3. Ciprofloxacin (CIP) 5 15 I 4. Cefoxitin (CX) 30 0 R 5. Amoxycillin/Clavulanic acid (AMC) 20/10 11 R 6. Tetracycline (TE) 30 18 I 7. Chloramphenicol (C) 30 30 S 8. Co-trimoxazole (COT) 25 16 I Dodeca Universal-XII DE027 9. Ofloxacin (OF) 5 16 I 10. Cefadroxil (CFR) 30 21 S 11. Doxycycline HCl (DO) 30 16 I 12. Cloxacillin (COX) 5 18 I 13. Azithromycin (AZM) 30 21 S 14. Cefotaxime (CTX) 10 20 S 15. Ceftriaxone (CTR) 30 20 S 16. Ticarcillin (TI) 75 17 I 17. Piperacillin/ Tazobactam (PIT) 100/10 25 S 18. Ciprofloxacin (CIP) 5 15 I 19. Levofloxacin (LE) 5 21 S 20. Ceftazidime (CAZ) 30 22 S R = resistant (≤ 14 mm); S = sensitive (≥ 20 mm); I = intermediate (15–19 mm). A hemolytic assay was conducted for the safety assessment, wherein the cells and cell-free supernatant of the strain did not exhibit detectable hemolytic activity. However, the 10-fold concentrated cell-free supernatant showed media color diffusion (yellow to green color) with no clear zone of inhibition, indicative of potential partial hemolysis (α hemolysis) or absence of hemolysis. Triton x100 was used as the positive control, displaying a clear zone around the well (β hemolysis). In contrast, Nisin (1.0 mg/mL) was the negative control, demonstrating no hemolytic activity (Fig. 7 D). Similar findings were reported for L. pentosus 22C, isolated from traditional yogurt, which also lacked hemolytic activity [ 93 ]. The absence of clear hemolytic (β) activities confirms that the strain is safe to use. Furthermore, the strain exhibited 23% auto-aggregation (Fig. 8 A) and 51% co-aggregation with S. Typhi (Fig. 8 B). Previous reports have indicated that L. pentosus exhibits notable co-aggregation abilities, including solid co-aggregation with Streptococcus mutans [ 94 ]. Many L. pentosus strains isolated from naturally fermented Aloreña table olives exhibited similar auto-aggregation and co-aggregation. Among them, 19% of L. pentosus strains had a high capability to auto-aggregate (50–77.92%), while 42% had a medium auto-aggregation capacity (35–50%). They also showed different ranges of co-aggregation with pathogens, including E. coli , Salmonella , Listeria innocua , and Staphylococcus aureus [ 80 ]. Moreover, the antibacterial activity of the cell-free supernatant (CFS) was evaluated using an agar-well diffusion assay, revealing antibacterial activity against various pathogens, including both Gram-positive and Gram-negative strains. Notably, in some instances, the antibacterial activity of the CFS was comparable to streptomycin (antibiotic) and superior to nisin (commercially available bacteriocin). Further details are provided in Table 5 . These observations are consistent with findings reported for L. pentosus strains provided by CICC (China Centre of Industrial Culture Collection) [ 95 ] and L. pentosus ZFM94, isolated from infant faces [ 82 ]. These evaluations collectively offer insights into various functional characteristics of L. pentosus , contributing to a comprehensive understanding of its probiotic attributes. Table 5 Represents the antibacterial activity of CFS of the isolated L. pentosus strain against different food pathogens. Sl No. Bacteria ZOI Cell-Free Supernatant in mm ZOI Nisin in mm ZOI Streptomycin in mm 1. Listeria monocytogenes 19 10 19 2. Vibrio harveyi 16 14 13 3. Streptococcus mutans 16 13 13 4. Staphylococcus aureus 16 10 (not clear) 16 5. Streptococcus thermophilus 18 0 14 6. Corynebacterium callunae 20 11 20 7. Enterococcus gallinarum 18 14 19 8. Vibrio cholerae 16 0 14 9. Bacillus cereus 18 13 25 10. Pseudomonas putida 16 0 19 11. Escherichia coli 16 0 20 12. Pseudomonas aeruginosa 16 10 16 13. Clostridium perfringens 21 0 16 14. Salmonella enterica subsp. enterica ser. Typhi 18 0 16 15. Salmonella enterica subsp. enterica ser. Paratyphi 17 0 17 ZOI is the zone of inhibition, and CFS is the cell-free supernatant of the isolated L. pentosus strain. Table 6: Antibacterial activity against various food pathogens. ZOI= Zone of Inhibition, CFS= Cell free supernatant. Sl No. Bacteria ZOI CFS in mm ZOI Nisin in mm ZOI Streptomycin in mm 1 Listeria monocytogenes 19 10 19 2 Vibrio harveyi 16 14 13 3 Streptococcus mutans 16 13 13 4 Staphylococcus aureus 16 10 (not clear) 16 5 Streptococcus thermophilus 18 0 14 6 Corynebacterium callunae 20 11 20 7 Enterococcus gallinarum 18 14 19 8 Vibrio cholerae 16 0 14 9 Bacillus cereus 18 13 25 10 Pseudomonas putida 16 0 19 11 Escherichia coli 16 0 20 12 Pseudomonas aeruginosa 16 10 16 13 Clostridium perfringens 21 0 16 14 Salmonella typhi 18 0 16 15 Salmonella paratyphi 17 0 17 Conclusions The genomic and probiotic characterization of the L. pentosus strain isolated from fermented rice revealed that the strain is a potential probiotic that can be used in the food industry. The genome study revealed that it carries genes that play a significant role in adhesion, aggregation, stress tolerance, antibiotic resistance, CRISPR CAS, and secondary metabolites, including bacteriocin. It also showed its phylogenetic relationship with L. plantarum . Probiotic characterization through different assays revealed the growth in various extreme situations, including low pH, the presence of bile salt, intestinal and gastric conditions, and their antioxidant potential. Additionally, the non-hemolysis and non-proteolytic natures were also demonstrated. Its broad range of antibacterial activity, as well as its antibiotic sensitivity and resistance, was also disclosed. These comprehensive findings collectively underscore the suitability of this strain for probiotic applications within the food industry, implying that it might be a promising prospect for functional food development and human health enhancement initiatives. Declarations Supplementary Materials: Figure S1: Illustrates the identification of biosynthetic gene clusters by antiSMASH 6.0 from the isolated L. pentosus genome. The predicted gene cluster showed a significant hit with RiPP-like and T3PKS clusters: a. Region 6.1 represents the RiPP-like cluster, and other unspecified ribosomally synthesized and post-translationally modified peptide products consist of two peptide bacteriocin clusters with ABC transporter and accessory gene; b. Region 32.1-T3PKS-Type III Polyketide synthase cluster with hydroxymethylglutaryl-CoA synthase region. Figure S2: Cluster Blast of RiPP region of L. pentosus in anti-SMASH shows the different genomes with similar bacteriocin clusters and their percentage of identity. Table S1: Sugar utilization pattern of isolated L. pentosus strain. Table S2: Represents the viability of the isolated L. pentosus strain in the presence of acid, bile salt, and gastric-intestinal conditions. Author Contributions: Conceptualization: R.K. and AC.; methodology: A.C.; writing—original draft preparation: A.C.; funding acquisition: R.K. All authors have read and agreed to the published version of the manuscript. Funding: This work was supported by the LSRB (The Life Sciences Research Board (LSRB) of Defense Research & Development Organization, GAP-537 and MOFPI (Ministry of Food Processing Industries, GAP-536). CSIR provides funding for researchers. We thank the Director, CSIR-CFTRI, Mysore, for providing the facilities. Data Availability Statement: The original contributions presented in the study are included in the article/Supplementary Material, further inquiries can be directed to the corresponding author. Conflicts of Interest: The authors declare no conflicts of interest. References Narzary Y, Brahma J, Brahma C et al (2016) A study on indigenous fermented foods and beverages of Kokrajhar, Assam, India. J. Ethn. Foods 3: 284–291. https://doi.org/10.1016/j.jef.2016.11.010. Rezac S, Kok CR, Heermann M et al (2018) Fermented foods as a dietary source of live organisms. Front. Microbiol. 9: 396129. https://doi.org/10.3389/fmicb.2018.01785. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {\"props\":{\"pageProps\":{\"initialData\":{\"identity\":\"rs-4845642\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":false,\"archivedVersions\":[],\"articleType\":\"Research Article\",\"associatedPublications\":[],\"authors\":[{\"id\":348453026,\"identity\":\"d713d590-74ea-42b0-a30b-6bbc82c88d0e\",\"order_by\":0,\"name\":\"Athira Cheruvari\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"CSIR-Central Food Technological Research Institute\",\"correspondingAuthor\":false,\"submittingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Athira\",\"middleName\":\"\",\"lastName\":\"Cheruvari\",\"suffix\":\"\"},{\"id\":348453027,\"identity\":\"0620a15f-bba9-479a-abff-62cf08735064\",\"order_by\":1,\"name\":\"Rajagopal Kammara\",\"email\":\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABB0lEQVRIie3RsUoDMRjA8YRAsny26xcC9wZCJFAc7k1crst1UThwPWxcdCnOSn0I38CDwnUpiNu53dG1QicXizSpi6B37eiQ/xII+fElhJBQ6B+G38tp1BeWENBRnxB6XRxA0MhJ4YmRllB7EBk+VYkjxBNCOomczl+XWY7Uvr039SrTeHw7c1Py+KyNqN75pbkvkdHpyJw8ao2DxdCRMr2wLSQCSBVw5EylXIHeXA0KR6id7SFfCFyW4hP8lJemmygQpTq6QQTknO1ItWeKnAAzD3eoEVImd2+p3JSk4y24EM0y+4jHz/OSrlcbf7FRU6/zuJX472O/N5PW4z5R/0FCoVAo9KMtD+pWwCpK2VoAAAAASUVORK5CYII=\",\"orcid\":\"\",\"institution\":\"CSIR-Central Food Technological Research Institute\",\"correspondingAuthor\":true,\"submittingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Rajagopal\",\"middleName\":\"\",\"lastName\":\"Kammara\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2024-08-02 04:52:29\",\"currentVersionCode\":1,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-4845642/v1\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-4845642/v1\",\"draftVersion\":[],\"editorialEvents\":[{\"content\":\"https://doi.org/10.1007/s12602-024-10378-1\",\"type\":\"published\",\"date\":\"2024-10-21T15:58:09+00:00\"}],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":63856229,\"identity\":\"17a491e2-75f6-4e13-9a86-4b8adbc43a67\",\"added_by\":\"auto\",\"created_at\":\"2024-09-03 05:25:28\",\"extension\":\"jpg\",\"order_by\":1,\"title\":\"Figure 1\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":77627,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e\\u003cstrong\\u003eA.\\u003c/strong\\u003e Antibacterial zone of inhibition against \\u003cem\\u003eS.\\u003c/em\\u003e Typhi; \\u003cstrong\\u003eB.\\u003c/strong\\u003e isolated bacterial colony (\\u003cem\\u003eL. pentosus\\u003c/em\\u003e) in MRS agar plate; \\u003cstrong\\u003eC\\u003c/strong\\u003e. Gram staining of isolated strain shows Gram-positive; \\u003cstrong\\u003eD\\u003c/strong\\u003e. SEM image of isolated strain (\\u003cem\\u003eL. pentosus\\u003c/em\\u003e).\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"1.jpg\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4845642/v1/680f28e571cae38b6d0384c2.jpg\"},{\"id\":63854505,\"identity\":\"287ed0dc-a9b1-46c0-bdc2-8509d983435f\",\"added_by\":\"auto\",\"created_at\":\"2024-09-03 05:01:28\",\"extension\":\"jpg\",\"order_by\":2,\"title\":\"Figure 2\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":45478,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eGrowth of isolated strain in different conditions: \\u003cstrong\\u003eA\\u003c/strong\\u003e. Growth in different media; \\u003cstrong\\u003eB.\\u003c/strong\\u003e Growth in MRS media with various pHs; \\u003cstrong\\u003eC\\u003c/strong\\u003e. Growth in MRS media with varying concentrations of NaCl; \\u003cstrong\\u003eD\\u003c/strong\\u003e. Growth in MRS media with different concentrations of phenol.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"2.jpg\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4845642/v1/26af3148f28e5f80d7b5839f.jpg\"},{\"id\":63854503,\"identity\":\"b60f65a8-285c-4894-ba78-667af3cad1a6\",\"added_by\":\"auto\",\"created_at\":\"2024-09-03 05:01:28\",\"extension\":\"jpg\",\"order_by\":3,\"title\":\"Figure 3\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":30092,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eRepresents a phylogenetic tree constructed using a 16S rRNA sequence (Maximum Likelihood method)\\u003cem\\u003e. \\u003c/em\\u003eThe test sample is highlighted as an isolated strain (\\u003cem\\u003eL. pentosus \\u003c/em\\u003estrain).\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"3.jpg\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4845642/v1/0cf381b20f540b77a209f371.jpg\"},{\"id\":63854510,\"identity\":\"56182474-b76f-47f4-a0c8-1d669b186352\",\"added_by\":\"auto\",\"created_at\":\"2024-09-03 05:01:28\",\"extension\":\"jpg\",\"order_by\":4,\"title\":\"Figure 4\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":270934,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e\\u003cstrong\\u003eA.\\u003c/strong\\u003e Depicts the circular genome map of the isolated \\u003cem\\u003eL. pentosus\\u003c/em\\u003egenerated using the CG view server tool. The blue region illustrates coding sequences (CDSs), while the grey region represents contigs. Moving from the outer to the inner rings, the features displayed include CDSs on the forward strand, CDSs on the reverse strand, and contigs. \\u003cstrong\\u003eB.\\u003c/strong\\u003e Illustrates various other genomic properties of \\u003cem\\u003eL. pentosus\\u003c/em\\u003e detected by CG view server; Alien hunter represents the prediction of putative Horizontal Gene Transfer (HGT) events; Phigaro detect prophage regions; Mobile OG-db detects mobile genetic elements (MGEs); VirSorter used for mining viral signal from microbial genomic data; CRISPR/Cas Finder identified CRISPR arrays and their associated Cas proteins; CARD detects antimicrobial resistance genes.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"4.jpg\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4845642/v1/8980393c176ff8c95b2ca9f1.jpg\"},{\"id\":63854507,\"identity\":\"a914f8e5-fe9c-468e-b9bc-4cee2e906523\",\"added_by\":\"auto\",\"created_at\":\"2024-09-03 05:01:28\",\"extension\":\"jpg\",\"order_by\":5,\"title\":\"Figure 5\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":364089,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e\\u003cstrong\\u003eA.\\u003c/strong\\u003eDepicts bacteriocin gene clusters of isolated \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strain identified using BAGEL 4; \\u003cstrong\\u003ea.\\u003c/strong\\u003e Pediocin cluster; \\u003cstrong\\u003eb.\\u003c/strong\\u003e Plantaricin E and F cluster, which is a 2-peptide bacteriocin with immunity genes, ABC transporter genes, and accessory genes. In addition, Lactococcin is also present in the same node; \\u003cstrong\\u003ec.\\u003c/strong\\u003eputative bacteriocin (Bovicin 255 variant). \\u003cstrong\\u003eB\\u003c/strong\\u003e. Illustrates\\u003cstrong\\u003e \\u003c/strong\\u003ethe prophage region in the isolated\\u003cem\\u003e L. pentosus \\u003c/em\\u003egenome identified using\\u003cem\\u003e PHASTER. \\u003c/em\\u003eThe green-color\\u003cem\\u003e \\u003c/em\\u003eregion in the genome shows a\\u003cem\\u003e \\u003c/em\\u003ecomplete hit with the virus and prophage database, and the red color indicates an incomplete score; \\u003cstrong\\u003ea.\\u003c/strong\\u003egenome view of prophage region and expanded genome view of prophage region in node 13; \\u003cstrong\\u003eb. \\u003c/strong\\u003egenome view of prophage region and expanded genome view of node 15 prophage region. \\u003cstrong\\u003eC\\u003c/strong\\u003e. CARD analysis results of\\u003cem\\u003e L. pentosus\\u003c/em\\u003egenome.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"5.jpg\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4845642/v1/20c9e03514fb5cff9ead4433.jpg\"},{\"id\":63856230,\"identity\":\"b5d5caa9-8ba6-492b-a29f-8ae467366883\",\"added_by\":\"auto\",\"created_at\":\"2024-09-03 05:25:28\",\"extension\":\"jpg\",\"order_by\":6,\"title\":\"Figure 6\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":44578,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e\\u003cstrong\\u003eA.\\u003c/strong\\u003e Represents the growth of the isolated \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strain in pH 3, 0.15% bile salt, gastric and intestinal conditions; \\u003cstrong\\u003eB\\u003c/strong\\u003e. Represents the percentage of survivability of the isolated \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strain in pH 3, 0.15% bile salt, gastric and intestinal conditions.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"6.jpg\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4845642/v1/a473a5e68c6b03dd417a1157.jpg\"},{\"id\":63854509,\"identity\":\"27c0591b-ce8c-444c-8c79-407b337368e2\",\"added_by\":\"auto\",\"created_at\":\"2024-09-03 05:01:28\",\"extension\":\"jpg\",\"order_by\":7,\"title\":\"Figure 7\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":44814,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eRepresents different probiotic assay plates: \\u003cstrong\\u003eA. \\u003c/strong\\u003eBile hydrolysis assay, cultures added in agar well and culture in direct bile agar; a,b—\\u003cem\\u003eB. adolescentis\\u003c/em\\u003e(positive control); c,d—\\u003cem\\u003eL. pentosus \\u003c/em\\u003e(Isolated strain as test); e,f—\\u003cem\\u003eS. \\u003c/em\\u003eTyphi\\u003cem\\u003e \\u003c/em\\u003e(negative control); \\u003cstrong\\u003eB\\u003c/strong\\u003e, \\u003cstrong\\u003eC\\u003c/strong\\u003e. Antibiotic susceptibility test—\\u003cstrong\\u003eB\\u003c/strong\\u003e. Universal-1 OD308; \\u003cstrong\\u003eC\\u003c/strong\\u003e. Dodeca Universal—XII DE027; \\u003cstrong\\u003eD.\\u003c/strong\\u003e Hemolytic assay; \\u003cstrong\\u003ea.\\u003c/strong\\u003eCFS concentrated (isolated \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strain CFS)—31 mm zone (yellow to green);\\u003cstrong\\u003eb.\\u003c/strong\\u003e CFS (isolated \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strain CFS)—11 mm zone (yellow to green); \\u003cstrong\\u003ec.\\u003c/strong\\u003e Nisin—no zone; d. Triton X 100—25 mm (clear zone; +ve control).\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"7.jpg\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4845642/v1/64e45cff004167f2b2196d1c.jpg\"},{\"id\":63854506,\"identity\":\"3a54f000-0981-4578-b94b-890d02e6a592\",\"added_by\":\"auto\",\"created_at\":\"2024-09-03 05:01:28\",\"extension\":\"jpg\",\"order_by\":8,\"title\":\"Figure 8\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":124896,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e\\u003cstrong\\u003eA.\\u003c/strong\\u003eIllustrates the auto-aggregation of the isolated \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strain by the Gram-staining method; \\u003cstrong\\u003eB\\u003c/strong\\u003e. illustrates the co-aggregation of the isolated \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strain with \\u003cem\\u003eS.\\u003c/em\\u003e Typhi. The Gram-positive \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strain is shown in blue, and the Gram-negative \\u003cem\\u003eS.\\u003c/em\\u003e Typhi strain in red.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"8.jpg\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4845642/v1/8787f47ec4f2b48fc4dc64e9.jpg\"},{\"id\":67682021,\"identity\":\"4ab766c2-557f-4d9a-a86e-637892916abf\",\"added_by\":\"auto\",\"created_at\":\"2024-10-28 16:12:36\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":2184706,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4845642/v1/da6ef2c7-ed10-4dc6-8c46-7974981db17f.pdf\"},{\"id\":63854512,\"identity\":\"bcfb0f07-a53a-4638-afe3-6662391e50ac\",\"added_by\":\"auto\",\"created_at\":\"2024-09-03 05:01:29\",\"extension\":\"doc\",\"order_by\":1,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"supplement\",\"size\":4531712,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"Athirasupplementary.doc\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4845642/v1/1a0f84d835d08c21e7150803.doc\"}],\"financialInterests\":\"No competing interests reported.\",\"formattedTitle\":\"Genomic Characterization and Probiotic Properties of Lactiplantibacillus pentosus Isolated from Fermented Rice\",\"fulltext\":[{\"header\":\"Introduction\",\"content\":\"\\u003cp\\u003eMany ethnic cultures consider fermented foods essential to their customs and traditions [\\u003cspan citationid=\\\"CR1\\\" class=\\\"CitationRef\\\"\\u003e1\\u003c/span\\u003e]. Fermentation has been used for centuries to extend the shelf life of food and improve its flavour, texture, and nutritional value [\\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2\\u003c/span\\u003e]. During fermentation, an organism converts a carbohydrate into an acid or alcohol. The most common yeast strain used in fermentation processes is \\u003cem\\u003eSaccharomyces cerevisiae\\u003c/em\\u003e, which is used to produce wines, beers, and ciders [\\u003cspan citationid=\\\"CR3\\\" class=\\\"CitationRef\\\"\\u003e3\\u003c/span\\u003e]. Lactic acid bacteria (LAB) are also associated with fermented foods, including \\u003cem\\u003eEnterococcus\\u003c/em\\u003e, \\u003cem\\u003eLactobacillus\\u003c/em\\u003e, \\u003cem\\u003eLactococcus\\u003c/em\\u003e, \\u003cem\\u003eLeuconostoc\\u003c/em\\u003e, \\u003cem\\u003ePediococcus\\u003c/em\\u003e, and \\u003cem\\u003eWeissella\\u003c/em\\u003e species. Additionally, cheese-derived \\u003cem\\u003eBifidobacterium\\u003c/em\\u003e and species of Bacillus in fermented legume-based foods are included in this category [\\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e4\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e5\\u003c/span\\u003e]. These microorganisms, consumed through food, play a crucial role in maintaining good health and are called pro-biotics [\\u003cspan citationid=\\\"CR6\\\" class=\\\"CitationRef\\\"\\u003e6\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eProbiotics have been used to modify the intestinal microbiota to improve the health of both humans and animals. To reduce the risk of gastrointestinal (GI) infections, several well-characterized lactobacilli and bifidobacterial strains are already available for human consumption [\\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e7\\u003c/span\\u003e]. The consumption of probiotics has been shown to improve intestinal health by regulating the microbiota, stimulating the development of the immune system, improving nutrient bioavailability, reducing the risk of various cancers, enhancing lactose digestion, preventing cardiovascular diseases, and lowering the risk of certain other diseases [\\u003cspan citationid=\\\"CR8\\\" class=\\\"CitationRef\\\"\\u003e8\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e9\\u003c/span\\u003e]. Probiotics are generally considered safe, although some LAB species have been linked to opportunistic infections. Further research is needed to determine the advantages and adverse effects of their use [\\u003cspan citationid=\\\"CR10\\\" class=\\\"CitationRef\\\"\\u003e10\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e11\\u003c/span\\u003e]. Therefore, when evaluating the general safety of probiotic-based foods and beverages, factors such as the source and nature of the probiotics, their pathogenicity, the method of administration, their potential to carry antibiotic resistance genes, the level of exposure, the host\\u0026rsquo;s health, and their intended use must be taken into account [\\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e12\\u003c/span\\u003e]. A microorganism\\u0026rsquo;s potential to serve as a probiotic depends on its pathogenicity, non-poisonous nature, allergenicity, carcinogenicity, or mutagenicity, and it should not have any adverse effects. It should be compatible with the food matrix and associated handling and storage requirements. It should be able to colonize the intestinal mucosa and survive through the gastrointestinal tract (adhesion characteristics, tolerance to acid and bile). Additionally, the host should benefit from it [\\u003cspan citationid=\\\"CR13\\\" class=\\\"CitationRef\\\"\\u003e13\\u003c/span\\u003e]. Furthermore, the strain must endure at the administration sites, multiply, and colonize for maximal activity in this particular spot. Moreover, the immune system should be able to tolerate it [\\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e7\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eIndia offers various region-specific fermented food products and beverages [\\u003cspan citationid=\\\"CR14\\\" class=\\\"CitationRef\\\"\\u003e14\\u003c/span\\u003e]. These traditional foods are typically prepared at home through natural fermentation. Despite their long history, many of these fermented foods have not been thoroughly studied from a scientific perspective. Therefore, utilizing modern food microbiology, such as sequence-based taxonomy, and examining the bioactive compounds present in these foods through metabolomics and other technologies, could help validate the value of this traditional food culture [\\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e15\\u003c/span\\u003e]. The Indian Himalayan Region is known for its traditional fermented foods and alcoholic beverages, making it a gastronomic hotspot. Alcoholic drinks have deep ceremonial significance for the ethnic communities of this region and have been consumed at cultural, social, and religious events for generations [\\u003cspan citationid=\\\"CR16\\\" class=\\\"CitationRef\\\"\\u003e16\\u003c/span\\u003e]. The main ingredients used in these foods include cereals, vegetables, meat, unripe fruits, legumes, and bamboo shoots [\\u003cspan citationid=\\\"CR17\\\" class=\\\"CitationRef\\\"\\u003e17\\u003c/span\\u003e]. Ethnic fermented foods, such as bhaturu, marchu, dosha, chilra, bedvin roti, seera, pinni, pakk, thuktal, sepubari, bari, churpa, and aska, are frequently made in Himachal Pradesh. Cereals are the main ingredient in these products. Himachal Pradesh is home to several traditional fermented alcoholic drinks made from millets, apples, wild apricots, and grapes, including sura, lugri, kinnauri, chakti, chulli, behmi, and ara [\\u003cspan citationid=\\\"CR16\\\" class=\\\"CitationRef\\\"\\u003e16\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003e \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strains are lactic acid bacteria (LAB) commonly found in various fermented olives from the Mediterranean region, with several strains exhibiting probiotic characteristics [\\u003cspan citationid=\\\"CR18\\\" class=\\\"CitationRef\\\"\\u003e18\\u003c/span\\u003e]. Due to its genetic diversity and adaptability, it possesses a range of molecular defenses to withstand harsh environmental conditions during processing and consumption [\\u003cspan citationid=\\\"CR19\\\" class=\\\"CitationRef\\\"\\u003e19\\u003c/span\\u003e]. Research indicates that \\u003cem\\u003eL. pentosus\\u003c/em\\u003e derived from vegetable fermentation offers numerous health benefits, including cholesterol reduction, antioxidant, anticancer, immunomodulatory activities, and antagonistic and antibacterial effects against gut pathogens [\\u003cspan citationid=\\\"CR20\\\" class=\\\"CitationRef\\\"\\u003e20\\u003c/span\\u003e]. The safety and probiotic properties of \\u003cem\\u003eL. pentosus\\u003c/em\\u003e, a GABA-producing strain isolated from Thai pickles, were evaluated through whole genome sequencing, demonstrating its safety and absence of transferable antimicrobial resistance (AMR) genes. The genome does not contain genes associated with biogenic amines, antimicrobial compounds, or clinically relevant toxins [\\u003cspan citationid=\\\"CR21\\\" class=\\\"CitationRef\\\"\\u003e21\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eProbiotic traits and health advantages vary according to the strain. Every strain is distinct. The primary goal of this study was to report on isolating probiotic \\u003cem\\u003eL. pentosus\\u003c/em\\u003e (strain krglsrbmofpi2) from fermented foods from unknown ethnic groups in Himachal Pradesh, India. Its genomic and probiotic properties were assessed to determine whether the isolated strain could be used as a safe food preservative or as a probiotic food.\\u003c/p\\u003e\"},{\"header\":\"Materials and Methods\",\"content\":\"\\u003cdiv id=\\\"Sec3\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eIsolation of Probiotic Bacteria\\u003c/h2\\u003e \\u003cp\\u003eVarious fermented foods, including fermented rice sourced from villages in Himachal Pradesh, were collected for investigation. From the collected samples, a 1.0% (1.0 mL) aliquot was mixed with 9 mL of peptone broth and incubated at 37\\u0026deg;C under aerobic conditions (200 RPM agitation in New Brunswick Innova 44, incubator shaker, Ham-burg, Germany.) for a duration of 24 h. Subsequently, the sample was spread-plated onto de Man, Rogosa, and Sharpe (MRS) agar plates to facilitate the isolation of various bacterial strains. \\u003cem\\u003eSalmonella enterica\\u003c/em\\u003e subsp. \\u003cem\\u003eenterica\\u003c/em\\u003e ser. Typhi (\\u003cem\\u003eS.\\u003c/em\\u003e Typhi) was used to evaluate the antibacterial activity of the isolated strains. The selected strain, active against \\u003cem\\u003eS.\\u003c/em\\u003e Typhi, was sub-cultured (1.0%) in MRS broth and incubated at 37\\u0026deg;C for 24 h, 200 RPM, in a shaker incubator. The nature of the antimicrobial compound was determined by testing its activity in the presence of different enzymes. The enzyme solutions (1 mg/mL) were prepared as follows: trypsin (in Tris-HCl 40 mM, pH 8.2), pepsin (in 0.002 M HCl, pH 2.0), chymotrypsin (in 50 mm Tris-HCl, pH 8.0), catalase (in 20 mM HCl, pH 2.0), protease (in water), lysozyme (in 10 mM Tris-HCl, pH 8.0), amylase (in water), lipase (in water). All enzymes were from Sigma-Aldrich (St. Louis, MO, USA). Then, 1.0 mL of cell-free supernatant (CFS) solution was added to 1.0 mL of the enzyme solutions. After 1 h of incubation at 37\\u0026deg;C, samples were boiled for 5 min at 100\\u0026deg;C to inactivate the enzyme. Concentrated CFS (60 \\u0026micro;L, Enzyme-treated CFS concentrated using a vacuum concentrator, Eppendorf, Hamburg, Germany) was used to estimate their antimicrobial activity through an agar-well diffusion assay against \\u003cem\\u003eS.\\u003c/em\\u003e Typhi [\\u003cspan additionalcitationids=\\\"CR23\\\" citationid=\\\"CR22\\\" class=\\\"CitationRef\\\"\\u003e22\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR24\\\" class=\\\"CitationRef\\\"\\u003e24\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec4\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003ePreliminary Characterization of Isolates\\u003c/h2\\u003e \\u003cp\\u003eGram staining and scanning electron microscopy analyses are standard techniques initially used to characterize isolated bacteria [\\u003cspan citationid=\\\"CR25\\\" class=\\\"CitationRef\\\"\\u003e25\\u003c/span\\u003e]. Next, a variety of biochemical tests were performed, such as the catalase test (using \\u003cem\\u003eS.\\u003c/em\\u003e Typhi as a positive control), sugar utilization tests, growth in various media, growth at different pH (2.0 to 10), growth at multiple concentrations of NaCl (0 to 10%) [\\u003cspan citationid=\\\"CR24\\\" class=\\\"CitationRef\\\"\\u003e24\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR26\\\" class=\\\"CitationRef\\\"\\u003e26\\u003c/span\\u003e] and varying concentrations of phenol (0.1\\u0026ndash;0.4%) [\\u003cspan citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e27\\u003c/span\\u003e]. This growth investigation used 1.0% overnight-grown cells and incubated for 24 h at 37\\u0026deg;C, 200 RPM. Culture grown in MRS broth was used as a control. The optical density (OD) was measured at 600 nm after 24 h [\\u003cspan citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e27\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR28\\\" class=\\\"CitationRef\\\"\\u003e28\\u003c/span\\u003e]. The following different media were used in the study: M17 Broth, Sabouraud Dextrose Broth, Hi-veg (SD), Brain Heart Infusion Broth (BHI), Nutrient Broth (NB), Luria Bertani Broth (LB), Yeast Extract Peptone-Dextrose Broth (YPD), Elliker Broth (ELK), Bifidobacterium broth (BB), Tryptone soya broth (SB), de Man Rogosa, and Sharpe Broth (MRS) (all media from Hi Media) [\\u003cspan citationid=\\\"CR29\\\" class=\\\"CitationRef\\\"\\u003e29\\u003c/span\\u003e]. Moreover, LAB was identified at the species level using additional biochemical tests like sugar utilization pattern and other properties by using the HiLacto identification kit (KB020, [\\u003cspan citationid=\\\"CR30\\\" class=\\\"CitationRef\\\"\\u003e30\\u003c/span\\u003e], Hi Carbo kit (KB009A/KB009B1) [\\u003cspan citationid=\\\"CR31\\\" class=\\\"CitationRef\\\"\\u003e31\\u003c/span\\u003e], Hi Bacillus Identification Kit (KB013). All kits are from HiMedia, Mumbai, India [\\u003cspan citationid=\\\"CR32\\\" class=\\\"CitationRef\\\"\\u003e32\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec5\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eDNA Extraction\\u003c/h2\\u003e \\u003cp\\u003eThermo Fisher Scientific\\u0026rsquo;s Gene JET Bacterial Genomic DNA Purification Kit was used to extract genomic DNA. The kit\\u0026rsquo;s instructions were meticulously followed. A nano-drop spectrophotometer was used to verify the amount and quality of genomic DNA (Thermo Fisher Scientific, Waltham, MA, USA) [\\u003cspan citationid=\\\"CR33\\\" class=\\\"CitationRef\\\"\\u003e33\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec6\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003ePhylogenetic Analysis\\u003c/h2\\u003e \\u003cp\\u003eThe identity of cultures was verified by sequencing 16S rRNA using bacterial universal primers 27F (5\\u0026prime;-AGAGTTTGATCCTGGCTCAG-3\\u0026prime;) and 1492R (5\\u0026prime;-GGTTACCTTGTT ACGACTT-3\\u0026prime;) [\\u003cspan citationid=\\\"CR34\\\" class=\\\"CitationRef\\\"\\u003e34\\u003c/span\\u003e]. To identify the nearest neighbour in the evolutionary tree, the 16S rRNA sequences of similar organisms were taken from the NCBI database (National Centre for Biotechnology Information) and compared with the sequences of isolated isolates. The Tamura-Nei model and the Maximum Likelihood technique were used to estimate the evolutionary history [\\u003cspan citationid=\\\"CR35\\\" class=\\\"CitationRef\\\"\\u003e35\\u003c/span\\u003e]. Comparative analyses of 10 nucleotide sequences enabled the identification of the closest relatives in the phylogenetic tree constructed using the Neighbour-joining (NJ) method with a p-distance model in MEGA X software (Molecular Evolutionary Genetics Analysis, version 10.0.0, \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://megasoftware.net\\u003c/span\\u003e\\u003cspan address=\\\"https://megasoftware.net\\\" targettype=\\\"URL\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e, accessed on 21 March 2019) [\\u003cspan citationid=\\\"CR36\\\" class=\\\"CitationRef\\\"\\u003e36\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec7\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eGenomic Characterization\\u003c/h2\\u003e \\u003cp\\u003eGenome Sequencing and Mapping\\u003c/p\\u003e \\u003cp\\u003eWhole genome sequencing of the isolate was performed by Sandor Speciality Di-agnostics (Hyderabad, India) using Illumina Hi Seq 1000 technology [\\u003cspan citationid=\\\"CR33\\\" class=\\\"CitationRef\\\"\\u003e33\\u003c/span\\u003e]. The results were submitted to the National Centre for Biotechnology Information (NCBI). Contigs containing genomic DNA were built using the SPAdes v. 3.13.1 assembler. The NCBI PGAP (Prokaryotic Genome Annotation Pipeline) contributed to the annotation [\\u003cspan citationid=\\\"CR37\\\" class=\\\"CitationRef\\\"\\u003e37\\u003c/span\\u003e]. With the help of the CG View server, a circular genome map was created (\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://cgview.ca/\\u003c/span\\u003e\\u003cspan address=\\\"https://cgview.ca/\\\" targettype=\\\"URL\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e, accessed on 14 December 2022) [\\u003cspan citationid=\\\"CR38\\\" class=\\\"CitationRef\\\"\\u003e38\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec8\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eBioinformatic Analysis\\u003c/h2\\u003e \\u003cp\\u003eDifferent bioinformatics tools were used to annotate and identify functional genes.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec9\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eProbiotic Factors\\u003c/h2\\u003e \\u003cp\\u003eProbiotic genes were identified using blast analysis by searching the entire genome sequence for sequence similarity [\\u003cspan citationid=\\\"CR25\\\" class=\\\"CitationRef\\\"\\u003e25\\u003c/span\\u003e]. The functional annotations of the genome were manually examined for the presence of the essential probiotic genes through prokka-generated annotation based on prior studies [\\u003cspan citationid=\\\"CR39\\\" class=\\\"CitationRef\\\"\\u003e39\\u003c/span\\u003e]. Various proteins involved in stress response, resistance, adhesion, aggregation, and immunomodulation were examined [\\u003cspan citationid=\\\"CR40\\\" class=\\\"CitationRef\\\"\\u003e40\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec10\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eBacteriocin and Secondary Metabolite\\u003c/h2\\u003e \\u003cp\\u003eBAGEL v.4.0, an online web server, was used to mine the antimicrobial peptides (bacteriocin) produced by bacteria. The assembled genome in FASTA format was uploaded and processed using the default settings in BAGEL v.4.0 (\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttp://bagel4.molgenrug.nl\\u003c/span\\u003e\\u003cspan address=\\\"http://bagel4.molgenrug.nl\\\" targettype=\\\"URL\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e, accessed on 14 December 2022) [\\u003cspan citationid=\\\"CR41\\\" class=\\\"CitationRef\\\"\\u003e41\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR42\\\" class=\\\"CitationRef\\\"\\u003e42\\u003c/span\\u003e]. Secondary metabolite production was analyzed using the bacterial versions of antiSMASH 6.0 (accessed on 14 December 2022), one of the most popular software programs for this task. It is based on profile hidden Markov models of genes typically found in specific Gene clusters [\\u003cspan citationid=\\\"CR43\\\" class=\\\"CitationRef\\\"\\u003e43\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR44\\\" class=\\\"CitationRef\\\"\\u003e44\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec11\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003ePlasmids\\u003c/h2\\u003e \\u003cp\\u003eThe genome of the isolated strain was examined for the presence of plasmids using the Plasmid Finder web tool version 2.0.1 (\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://cge.food.dtu.dk/services/PlasmidFinder/\\u003c/span\\u003e\\u003cspan address=\\\"https://cge.food.dtu.dk/services/PlasmidFinder/\\\" targettype=\\\"URL\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e, accessed on 14 December 2022). A minimum coverage of 60% and a minimum identity percentage of 90% were added to the parameters. Plasmid Finder detects replicons in WGS and assigns the plasmid under study to incompatibility (Inc.) groups, referring to the plasmid\\u0026rsquo;s GenBank accession number as the reference for that group [\\u003cspan additionalcitationids=\\\"CR46\\\" citationid=\\\"CR45\\\" class=\\\"CitationRef\\\"\\u003e45\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR47\\\" class=\\\"CitationRef\\\"\\u003e47\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec12\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eCRISPR/CRISPR-Cas\\u003c/h2\\u003e \\u003cp\\u003eThe CRISPR regions were detected with a CRISPR online detection tool, CRISPR Cas finder (\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://crisprcas.i2bc.paris-saclay.fr/CrisprCasFinder/\\u003c/span\\u003e\\u003cspan address=\\\"https://crisprcas.i2bc.paris-saclay.fr/CrisprCasFinder/\\\" targettype=\\\"URL\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e, accessed on 14 December 2022). CRISPR refers to a specific family of tandem repeats in various bacterial genomes. CRISPR-Cas9 is a prokaryote adaptive immune system that protects cells from DNA virus infections [\\u003cspan citationid=\\\"CR48\\\" class=\\\"CitationRef\\\"\\u003e48\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec13\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eProphage Regions\\u003c/h2\\u003e \\u003cp\\u003eThe PHAge Search Tool Enhanced Release (PHASTER) web server (\\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ewww.phaster.ca\\u003c/span\\u003e\\u003cspan address=\\\"http://www.phaster.ca\\\" targettype=\\\"URL\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e, accessed on 14 December 2022) identified potential prophage locations. The PHASTER finds, annotates, and visually shows prophage sequences found in bacterial genomes or plasmids. Bacteriophage genes, both functional and non-functional, may be found in more than 20% of bacterial genomes. Prophage sequences can explain a significant portion of the variance found among bacterial species or clades. Prophage sequences may also aid in developing pathogenicity, antibiotic resistance, exploration of novel ecological niches, and strengthened adhesion in certain bacteria [\\u003cspan additionalcitationids=\\\"CR50\\\" citationid=\\\"CR49\\\" class=\\\"CitationRef\\\"\\u003e49\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR51\\\" class=\\\"CitationRef\\\"\\u003e51\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec14\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003ePathogenicity\\u003c/h2\\u003e \\u003cp\\u003eHuman pathogenicity and virulence were predicted by the Pathogen Finder v.1.1 website (accessed on 15 December 2022, from \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://cge.cbs.dtu.dk/services/PathogenFinder/\\u003c/span\\u003e\\u003cspan address=\\\"https://cge.cbs.dtu.dk/services/PathogenFinder/\\\" targettype=\\\"URL\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e). It is a web server that predicts the pathogenicity of bacteria by analyzing the proteome, genome, or raw data given by the user [\\u003cspan citationid=\\\"CR52\\\" class=\\\"CitationRef\\\"\\u003e52\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec15\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eAntibiotic Resistance Genes\\u003c/h2\\u003e \\u003cp\\u003eAntibiotic resistance genes were found using the Comprehensive Antibiotic Research Database (CARD Database, \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://card.mcmaster.ca/analyze\\u003c/span\\u003e\\u003cspan address=\\\"https://card.mcmaster.ca/analyze\\\" targettype=\\\"URL\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e, accessed on 16 December 2022). It contains bioinformatic tools for identifying antibiotic resistance genes in whole genome or partial genome sequence data, including unannotated raw sequence assembly contigs. It is a robustly curated database in a user-friendly format that assembles over 1600 known antibiotic resistance genes, allowing sophisticated antibiotic resistance analysis and query in a way that will benefit the broader biomedical research community [\\u003cspan additionalcitationids=\\\"CR54\\\" citationid=\\\"CR53\\\" class=\\\"CitationRef\\\"\\u003e53\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR55\\\" class=\\\"CitationRef\\\"\\u003e55\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec16\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eProbiotic Properties\\u003c/h2\\u003e \\u003cdiv id=\\\"Sec17\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003eAcid and Bile Tolerance\\u003c/h2\\u003e \\u003cp\\u003eFollowing a 24 h incubation period in MRS broth, the isolated strain was sub-cultured at a concentration of 10% in three different growth media: MRS broth adjusted to pH 3.0 to assess acid tolerance, MRS broth supplemented with 0.15% bile salt to evaluate bile tolerance, and standard MRS broth serving as the control (initial OD\\u0026mdash;0.5). The adjustment of pH to 3.0 was achieved using 1.0 N HCl. Cultures were maintained aerobically (200 RPM agitation) at 37\\u0026deg;C throughout the experiment [\\u003cspan additionalcitationids=\\\"CR57\\\" citationid=\\\"CR56\\\" class=\\\"CitationRef\\\"\\u003e56\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR58\\\" class=\\\"CitationRef\\\"\\u003e58\\u003c/span\\u003e]. Samples were withdrawn at intervals of 0, 3, 6, 9, 12, and 24 h for evaluation. Growth dynamics were assessed by measuring optical density at 600 nm (OD600) using a spectrophotometer (DLAB SP-V1000, Los Angeles, CA, USA). Additionally, serial dilutions of the cultures were plated onto MRS agar plates, followed by incubation for 24 h at 37\\u0026deg;C. Colony-forming units per millilitre (CFU/mL) were enumerated to quantify bacterial growth. All counts were obtained in duplicate and expressed as mean\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;SD [\\u003cspan citationid=\\\"CR59\\\" class=\\\"CitationRef\\\"\\u003e59\\u003c/span\\u003e]. Finally, the acid and bile tolerance/percentage of survivability was estimated by the following equation: Bile/Acid Tolerance (%) = [OD600 of test/OD600 of control)] \\u0026times; 100 [\\u003cspan additionalcitationids=\\\"CR61\\\" citationid=\\\"CR60\\\" class=\\\"CitationRef\\\"\\u003e60\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR62\\\" class=\\\"CitationRef\\\"\\u003e62\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec18\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eBile Salt Hydrolase Assay\\u003c/h2\\u003e \\u003cp\\u003eBile salt hydrolase (BSH) plays a crucial role in the metabolism of bile acids, particularly in the intestine. This enzyme facilitates the hydrolysis of bile acids conjugated with glycine and taurine, liberating free bile acids and amino acid residues. BSH activity of intestinal bacteria is one of the indirect ways of decreasing the cholesterol level in the human body [\\u003cspan citationid=\\\"CR63\\\" class=\\\"CitationRef\\\"\\u003e63\\u003c/span\\u003e]. The BSH activity of the isolated strain was detected using a plate as-say, with slight modifications. Using a sterile cork borer (6.0 mm diameter), wells were made on a bile salt agar (HiMedia, Mumbai, India) plate, and the fresh overnight cultures (100 \\u0026micro;L) were added. Plates were incubated in an aerobic environment for 24 h at 37\\u0026deg;C. MRS agar plates without bile salt were used as a control. The presence of precipitated bile acid (cholic acid) around colonies (opaque halo) or the formation of opaque granular white colonies was considered a positive reaction. Each sample was analyzed twice [\\u003cspan additionalcitationids=\\\"CR65\\\" citationid=\\\"CR64\\\" class=\\\"CitationRef\\\"\\u003e64\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR66\\\" class=\\\"CitationRef\\\"\\u003e66\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec19\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eLysozyme Tolerance\\u003c/h2\\u003e \\u003cp\\u003eLysozyme tolerance was assessed with a 24 h growth culture, washed with phosphate buffer saline (PBS) solution (NaCl\\u0026mdash;1.37 M, KCl\\u0026mdash;27 mM, Na2HPO4\\u0026mdash;10 mM, KH2PO4\\u0026mdash;18 mM, pH 7.0), resuspended the 1.0 mL of collected cells in 10 mL of PBS solution containing 0.01% lysozyme (Sigma-Aldrich, St. Louis, MO, USA), and incubated at 37\\u0026deg;C for 1 h. Subsequently, cell viability was estimated as CFU/mL by serially diluted cells plated on an MRS agar plate. All counts were obtained in duplicate, and mean values were taken [\\u003cspan citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e27\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR67\\\" class=\\\"CitationRef\\\"\\u003e67\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec20\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eGastrointestinal Tolerance\\u003c/h2\\u003e \\u003cp\\u003eIsolated strains were cultured for 24 h in MRS broth at 37\\u0026deg;C, and the absorbance at 600 nm (OD600) was measured using a spectrophotometer after incubation. Cells were pelletized by centrifugation (6000 RPM for 10 min at room temperature) and re-suspended in sterile MRS broth. Culture at 10% (0.5 OD) was added to MRS broth containing stimulated gastric fluid (0.3% pepsin at pH 3), intestinal fluid (0.1% pancreatin and 0.15% bovine bile salt at pH 8), and culture in MRS broth was used as control [\\u003cspan citationid=\\\"CR57\\\" class=\\\"CitationRef\\\"\\u003e57\\u003c/span\\u003e]. These cell suspensions were incubated for 24 h at 37\\u0026deg;C, 200 RPM. During the incubation, absorbance (OD at 600 nm) and the viable cell counts (CFU/mL) were determined for 0, 3, 6, 9, 12, and 24 h. The experiments were repeated twice, and the mean values were taken [\\u003cspan citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e27\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR68\\\" class=\\\"CitationRef\\\"\\u003e68\\u003c/span\\u003e]. The following equation measured the percentage of bacterial survival: Percentage of Survivability/Tolerance = (Growth of Test/Growth of Control) \\u0026times; 100 [\\u003cspan citationid=\\\"CR69\\\" class=\\\"CitationRef\\\"\\u003e69\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec21\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eCell Surface Hydrophobicity\\u003c/h2\\u003e \\u003cp\\u003eCell surface hydrophobicity refers to the tendency of microbial cell surfaces to interact with hydrophobic molecules or surfaces. It plays a significant role in microbial processes, including adhesion to host tissues, biofilm formation, and interactions with environmental surfaces. The measurement of cell surface hydrophobicity is crucial in understanding the behaviour and physiology of microorganisms. The overnight culture of the isolated strain was pelletized by centrifugation (6000 RPM, 10 min), washed twice using sterile phosphate buffer saline (PBS), and re-suspended in 3.0 mL of the same buffer. The initial absorbance (A0) was adjusted to 0.5 OD at 600nm. Then, 1.0 mL of xylene (Merck, Darmstadt, Germany)/chloroform/hexadecane (HiMedia, Mumbai, India) was added to the bacterial suspension and vortexed for 2 min [\\u003cspan citationid=\\\"CR63\\\" class=\\\"CitationRef\\\"\\u003e63\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR70\\\" class=\\\"CitationRef\\\"\\u003e70\\u003c/span\\u003e]. The mixture was subjected to phase separation by incubating at 37\\u0026deg;C for 1 h, and the absorbance of the aqueous phase (A1) was measured using a spectrophotometer at 600 nm. The experiments were repeated twice, and the mean values were taken [\\u003cspan citationid=\\\"CR28\\\" class=\\\"CitationRef\\\"\\u003e28\\u003c/span\\u003e]. The following equation measured the percentage of hydrophobicity:\\u003c/p\\u003e \\u003cp\\u003e% Cell surface hydrophobicity = (1\\u0026thinsp;\\u0026minus;\\u0026thinsp;A1/A0) \\u0026times; 100.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec22\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eDPPH (2,2-Diphenyl-1-picrylhydrazyl) Free Radical Scavenging Assay\\u003c/h2\\u003e \\u003cp\\u003eThe DPPH assay was used to measure the capacity to scavenge free radicals. The overnight culture was centrifuged for 15 min at room temperature at 6000 RPM, and the cell-free supernatant (CFS) was collected in a separate tube. CFS (2.0 mL) was mixed with 2,2-diphenyl-1-picrylhydrazyl (2.0 mL, DPPH\\u0026mdash;6 mg/100 mL in methanol). Methanol served as the blank, while 2.0 mL of DPPH in 2.0 mL of methanol served as the control. The reaction mixture was thoroughly vortexed and allowed to sit in the dark at 37˚C for 30 min. Absorbance was recorded at 517 nm [\\u003cspan citationid=\\\"CR71\\\" class=\\\"CitationRef\\\"\\u003e71\\u003c/span\\u003e]. Ascorbic acid (10 \\u0026micro;g/mL, Sigma-Aldrich, St. Louis, MO, USA) was used as a positive control in distilled water. Each sample analysis was conducted in duplicate to ensure accuracy and reliability, and the following formula was applied to determine the antioxidant activity:\\u003c/p\\u003e \\u003cp\\u003eFree radical scavenging activity (%) = (Ac\\u0026thinsp;\\u0026minus;\\u0026thinsp;As)/Ac \\u0026times; 100\\u003c/p\\u003e \\u003cp\\u003eAs is the sample absorbance, and Ac is the control absorbance at 517 nm [\\u003cspan citationid=\\\"CR60\\\" class=\\\"CitationRef\\\"\\u003e60\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cdiv id=\\\"Sec23\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003eProteolytic Activity Assays\\u003c/h2\\u003e \\u003cp\\u003eThe isolated strain was screened for proteolytic activity by an agar-well diffusion method on skim milk containing 2.0% (w/v) agar (Hi-Media). Actively growing culture (60 \\u0026micro;L) and CFS (60 \\u0026micro;L) were added to the wells made in an agar plate (skim milk agar) and incubated at 37\\u0026deg;C. Trypsin (1.0 mg/mL) was used as a positive control. A clear zone around the agar-well-containing bacteria indicates a positive result. This methodology enabled the qualitative assessment of the strain\\u0026rsquo;s ability to hydrolyse proteins, as evidenced by the formation of clear zones resulting from casein degradation within the skim milk agar medium [\\u003cspan citationid=\\\"CR72\\\" class=\\\"CitationRef\\\"\\u003e72\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec24\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eAntibiotic Susceptibility Testing\\u003c/h2\\u003e \\u003cp\\u003eThe antibiotic susceptibilities of isolated bacteria were determined by the disc diffusion method, as recommended by CLSI (Clinical and Laboratory Standards Institute, Wayne, PA, USA). Thus, 100 \\u0026micro;L of 0.5 OD culture mixed with MRS agar was plated. Antibiotic discs Universal-1 OD308 and Dodeca Universal-XII DE027 (HiMedia, Mumbai, India) were placed on the agar plates with culture. They were incubated at 37\\u0026deg;C for 24 h. The diameter of zones was measured, and the isolates were identified as sensitive, intermediate, or resistant according to CLSI tables and guidelines. The experiments were repeated twice [\\u003cspan citationid=\\\"CR73\\\" class=\\\"CitationRef\\\"\\u003e73\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cdiv id=\\\"Sec25\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003eHemolytic Activity\\u003c/h2\\u003e \\u003cp\\u003eThe isolated bacteria\\u0026rsquo;s hemolytic activity was determined by adding the 24 h grown culture and cell-free supernatant to the wells on blood agar plates containing 5.0% (w/v) sheep blood and incubating at 37\\u0026deg;C for 24 h. After incubation, the plates were examined for β-hemolysis (Clear zone of inhibition), α-hemolysis (Greenish zone), and non-hemolytic activities (γ-hemolysis). Triton X 100 (HiMedia, Mumbai, India) and Nisin (1 mg/mL, HiMedia, Mumbai, India) were used as controls. Each sample was examined twice. [\\u003cspan citationid=\\\"CR60\\\" class=\\\"CitationRef\\\"\\u003e60\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec26\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003eAuto-Aggregation and Co-Aggregation\\u003c/h2\\u003e \\u003cp\\u003eThe ability of isolates to auto-aggregate was evaluated by 24 h grown culture. OD at 600 nm was adjusted to 0.5, and cells were centrifuged (6000 RPM, 10 min), and pellets were resuspended in a standard saline solution. The absorbance was measured at 0 h and 3 h by a spectrophotometer at 600 nm without shaking the cell suspension [\\u003cspan citationid=\\\"CR74\\\" class=\\\"CitationRef\\\"\\u003e74\\u003c/span\\u003e]. The au-to-aggregation was estimated as follows:\\u003c/p\\u003e \\u003cp\\u003eAuto-aggregation (%) = (1\\u0026thinsp;\\u0026minus;\\u0026thinsp;At/A0) \\u0026times; 100, A0 is the initial absorbance, and At is the absorbance at 3 h [\\u003cspan citationid=\\\"CR75\\\" class=\\\"CitationRef\\\"\\u003e75\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eThe isolate\\u0026rsquo;s co-aggregation with pathogenic bacteria was evaluated by mixing pathogenic bacteria (3.0 mL) and probiotic cells (3.0 mL) in a sterile tube. Both cells had 0.5 OD (at 600 nm). We mixed it well and incubated it at 37\\u0026deg;C in static conditions. The absorbance was measured at OD 600 nm after 3 h. Pathogenic and probiotic bacteria were cultured separately, and their absorbance was taken as control. The percentage of co-aggregation was calculated according to the formula:\\u003c/p\\u003e \\u003cp\\u003eCo-aggregation (%) = [(Ax\\u0026thinsp;+\\u0026thinsp;Ay)/2\\u0026thinsp;\\u0026minus;\\u0026thinsp;Axy]/ [(Ax\\u0026thinsp;+\\u0026thinsp;Ay)/2] \\u0026times; 100, where Ax\\u0026mdash;represents the absorbance of the probiotic strain, Ay\\u0026mdash;represents the absorbance of the pathogenic bacteria under study, Axy\\u0026mdash;represents the absorbance of the mixture of both [\\u003cspan citationid=\\\"CR76\\\" class=\\\"CitationRef\\\"\\u003e76\\u003c/span\\u003e]. The food pathogen \\u003cem\\u003eS.\\u003c/em\\u003e Typhi was the pathogen used for the co-aggregation assay. Co-aggregated cells and auto-aggregated cells after 3 h of incubation were visualized under microscopy using Gram staining [\\u003cspan citationid=\\\"CR77\\\" class=\\\"CitationRef\\\"\\u003e77\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec27\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003eAntibacterial Activity\\u003c/h2\\u003e \\u003cp\\u003eThe antibacterial activity of the LAB strains against food pathogens was determined using the agar well diffusion assay. The pathogenic indicator bacteria used for the study included \\u003cem\\u003eSalmonella enterica\\u003c/em\\u003e subsp. \\u003cem\\u003eenterica\\u003c/em\\u003e ser. Typhi (new isolate), \\u003cem\\u003eSalmonella enterica\\u003c/em\\u003e subsp. \\u003cem\\u003eenterica\\u003c/em\\u003e ser. \\u003cem\\u003eParatyphi\\u003c/em\\u003e (MTCC 735), \\u003cem\\u003eListeria monocytogenes\\u003c/em\\u003e (MTCC 839), \\u003cem\\u003eStreptococcus mutants\\u003c/em\\u003e (MTCC 497), \\u003cem\\u003eStreptococcus thermophilus\\u003c/em\\u003e (New isolate), \\u003cem\\u003eStaphylococcus aureus\\u003c/em\\u003e (MTCC 1430), \\u003cem\\u003ePseudomonas aeruginosa\\u003c/em\\u003e (MTCC 1934), \\u003cem\\u003ePseudomonas putida\\u003c/em\\u003e (MTCC 2492), \\u003cem\\u003eVibrio cholerae\\u003c/em\\u003e (MTCC 3904), \\u003cem\\u003eVibrio harveyi\\u003c/em\\u003e (MTCC 7954), \\u003cem\\u003eCorynebacterium callunae\\u003c/em\\u003e (MTCC 700), \\u003cem\\u003eEnterococcus gallinarum\\u003c/em\\u003e (MTCC 7049), \\u003cem\\u003eClostridium perfringens\\u003c/em\\u003e (MTCC 450), \\u003cem\\u003eEscherichia coli\\u003c/em\\u003e (k2 strain) and \\u003cem\\u003eBacillus cereus\\u003c/em\\u003e (MTCC 430) [\\u003cspan citationid=\\\"CR78\\\" class=\\\"CitationRef\\\"\\u003e78\\u003c/span\\u003e]. For the well-diffusion assay, 100 \\u0026micro;L of an overnight culture of the indicator strain was mixed with BHI soft agar (0.75% agar in BHI Broth) and poured into a BHI agar plate. A 6.0 mm diameter well was made in the agar, and 60 \\u0026micro;L (1.0 mg/mL protein concentration of protein present in CFS) of 10-fold concentrated cell-free supernatant was added, and incubated the plates for 12 h to measure the zone of inhibition. The experiments were repeated twice [\\u003cspan citationid=\\\"CR79\\\" class=\\\"CitationRef\\\"\\u003e79\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec28\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eStatistical Analysis\\u003c/h2\\u003e \\u003cp\\u003eThe statistical data analysis was conducted using GraphPad Prism version 8.0.2 for Windows. All data were expressed as mean\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;standard deviation. One-way analysis of variance (ANOVA) was employed to compare a single parameter across multiple groups. A significance threshold of p\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05 was utilized, where statistical significance was determined.\\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"Results and Discussion\",\"content\":\"\\u003cdiv id=\\\"Sec30\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003ePreliminary Characterization of LAB Isolates\\u003c/h2\\u003e \\u003cp\\u003eThe strains isolated from fermented rice underwent initial characterization through physiological and biochemical testing. A diverse array of both Gram-positive and Gram-negative strains were obtained. Subsequent antibacterial assays against \\u003cem\\u003eS.\\u003c/em\\u003e Typhi revealed that only one strain exhibited significant activity, as evidenced by the observed zone of inhibition (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eA).\\u003c/p\\u003e \\u003cp\\u003eThe antimicrobial activity present in the cell-free supernatant (CFS) was subjected to enzymatic treatment with various enzymes, including trypsin, chymotrypsin, pro-tease, catalase, lysozyme, amylase, and lipase. Upon treatment, a decrease in antimicrobial activity was observed in the presence of protease, chymotrypsin, and trypsin, suggesting a proteinaceous nature of the antimicrobial compound. The zone of inhibition was measured at 16 mm for protease, 15 mm for chymotrypsin, and 18 mm for trypsin, while all other enzymes resulted in a consistent zone of inhibition of 19 mm. These findings underscore the susceptibility of the antimicrobial activity to enzymatic degradation by proteolytic enzymes, indicating its dependence on proteinaceous components for antimicrobial efficacy. Such insights are crucial for elucidating the biochemical nature of the antimicrobial compounds present in the CFS.\\u003c/p\\u003e \\u003cp\\u003eMorphological examination indicated that colonies of this active strain were circular, smooth, and elevated (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eB), with Gram staining confirming this Gram-positive nature (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eC). Scanning electron microscopy (SEM) analysis further corroborated the rod-shaped morphology of the strain (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eD).\\u003c/p\\u003e \\u003cp\\u003eFurther characterization included catalase testing, which indicated that the isolated strain was catalase-negative, contrasting with the catalase-positive \\u003cem\\u003eS.\\u003c/em\\u003e Typhi used as a control. Growth assessment under varied conditions encompassed different culture media, pH levels, salt concentrations (NaCl), and phenol concentrations. Among the media tested, MRS broth supported the most rapid growth of the isolated strain, with significantly lower growth observed in other media (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003eA). Optimal growth occurred at pH 7.0 at 37\\u0026deg;C (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003eB), while exposure to increasing NaCl concentrations resulted in a progressive reduction in growth, with a notable decrease observed from 6.0% NaCl (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003eC). The observed characteristics\\u0026rsquo; parallel findings reported in \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strains were isolated from naturally fermented Alore\\u0026ntilde;a green table olives [\\u003cspan citationid=\\\"CR80\\\" class=\\\"CitationRef\\\"\\u003e80\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eEvaluation of the strain\\u0026rsquo;s tolerance to phenol revealed its ability to survive up to a 0.4% phenol concentration; however, it had diminished growth compared to control conditions, particularly at higher phenol concentrations (the growth decreased with increasing phenol concentration from 0.1 to 0.4% compared to the control) (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003eD). These findings underscore the adaptability and resilience of the isolated strain to various environmental conditions.\\u003c/p\\u003e \\u003cp\\u003eThe biochemical analysis indicated that the isolated bacteria could utilize all tested sugars, as outlined in Supplementary Table \\u003cspan refid=\\\"MOESM1\\\" class=\\\"InternalRef\\\"\\u003eS1\\u003c/span\\u003e. It includes lactose, sucrose, fructose, glucose, sorbitol, mannitol, etc. Consistently, \\u003cem\\u003eL. pentosus\\u003c/em\\u003e TEZU174 also exhibited a comparable sugar utilization profile [\\u003cspan citationid=\\\"CR81\\\" class=\\\"CitationRef\\\"\\u003e81\\u003c/span\\u003e]. Additionally, the results of various biochemical reactions, such as Voges-Proskauer\\u0026rsquo;s test, citrate utilization, ONPG, and nitrate reduction, are detailed in Table\\u0026nbsp;\\u003cspan refid=\\\"Tab7\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e.\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab1\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 1\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eBiochemical test results of the \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strain.\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"4\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eSl No.\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eTest\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003ePrinciple\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eResults\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e1.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eMalonate\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eMalonate utilization\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e\\u0026minus;ve\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e2.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eVoges Proskauer\\u0026rsquo;s\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eDetects acetoin production\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e\\u0026minus;ve\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e3.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eCitrate\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eCitrate utilisation\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e\\u0026minus;ve\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e4.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eONPG\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eDetects Beta galactosidase\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e+ve\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e5.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eNitrate Reduction\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eDetects Nitrate reduction\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e\\u0026minus;ve\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e6.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eCatalase\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eDetects Catalase activity\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e\\u0026minus;ve\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e7.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eArginine\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eArginine utilisation\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e\\u0026minus;ve\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e8.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eEsculin Hydrolysis\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eDetects Esculin Hydrolysis activity\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e+ve\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003ctfoot\\u003e \\u003ctr\\u003e\\u003ctd colspan=\\\"4\\\"\\u003e+ve: indicates positive results; \\u0026minus;ve: negative results.\\u003c/td\\u003e\\u003c/tr\\u003e \\u003c/tfoot\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003eConfirmation of its classification as a Lactiplantibacillus species was attained through 16S rRNA gene sequencing. Phylogenetic analysis unveiled a striking 99.6% similarity with strains, such as \\u003cem\\u003eLactiplantibacillus pentosus\\u003c/em\\u003e LMEM (MK240372.1), \\u003cem\\u003eLactiplantibacillus pentosus\\u003c/em\\u003e GCHI (MK245998.1), and several \\u003cem\\u003eLactiplantibacillus plantarum\\u003c/em\\u003e strains, including \\u003cem\\u003eLactiplantibacillus plantarum\\u003c/em\\u003e SCHI (MK246005.1), and \\u003cem\\u003eLactiplantibacillus plantarum\\u003c/em\\u003e MA8-6 (MG755354.1) (refer to Table\\u0026nbsp;\\u003cspan refid=\\\"Tab8\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e and Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab2\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 2\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eMicrobial identification using 16S rRNA-based molecular method. Sequences of strains having significant alignments are given in the table.\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"4\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eSl No.\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eDescription\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eIdentity\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eAccession No.\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e1.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eLactiplantibacillus plantarum\\u003c/em\\u003e strain SCHI\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e99.63%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMK246005.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e2.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eLactiplantibacillus pentosus\\u003c/em\\u003e strain GCHI\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e99.63%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMK245998.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e3.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eLactiplantibacillus pentosus\\u003c/em\\u003e strain LMEM\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e99.63%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMK240372.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e4.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eLactiplantibacillus plantarum\\u003c/em\\u003e strain MA8-6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e99.63%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMG755354.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e5.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eLactiplantibacillus plantarum\\u003c/em\\u003e strain NWAFU1580\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e99.63%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMK045823.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e6.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eLactiplantibacillus plantarum\\u003c/em\\u003e strain TTF18\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e99.63%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMK028367.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e7.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eLactiplantibacillus plantarum\\u003c/em\\u003e strain TTF17\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e99.63%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMK028366.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e8.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eLactiplantibacillus plantarum\\u003c/em\\u003e strain TTF16\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e99.63%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMK028365.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e9.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eLactiplantibacillus plantarum\\u003c/em\\u003e strain TTF15\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e99.63%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMK028364.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e10.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eLactiplantibacillus plantarum\\u003c/em\\u003e strain TTF14\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e99.63%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMK028363.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003eFurther confirmation of its identity as \\u003cem\\u003eLactiplantibacillus pentosus\\u003c/em\\u003e (formerly known as \\u003cem\\u003eLactobacillus pentosus\\u003c/em\\u003e) strains was achieved through whole genome sequencing. Both datasets, including 16S rRNA gene accession (MN165450.1) and WGS GenBank assembly accession (GCA_009295675.1), were duly submitted to NCBI.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec31\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eGenomic Characterization\\u003c/h2\\u003e \\u003cp\\u003eThe genomic analysis of \\u003cem\\u003eLactiplantibacillus pentosus\\u003c/em\\u003e reveals a composite structure consisting of 55 contigs (667,623 base pairs), with a total size of 3.7 Mb and a GC content of 46%. Further, 3342 coding sequences (CDSs) are identified within this genome, of which 3192 encode proteins. Comprehensive annotation further discloses the presence of 76 RNA genes, comprising 65 transfer RNA (tRNA), 7 ribosomal RNA (rRNA), and 4 non-coding RNA (ncRNA) genes.\\u003c/p\\u003e \\u003cp\\u003eFigure \\u003cspan refid=\\\"Fig4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003e, generated through the CG View Server, provides a visual representation of the genome map, elucidating the arrangement of coding sequences on the strand, contigs, GC content, GC skew, and other pertinent genomic attributes. Additionally, supplementary tools enhance the analysis by identifying various genomic elements and properties. Specifically, Alien Hunter predicts putative Horizontal Gene Transfer (HGT) events, Phigaro detects prophage regions, Mobile OG-db identifies mobile genetic elements (MGEs), VirSorter discerns viral signals within microbial genomic data, CRISPR/Cas Finder detects CRISPR arrays along with their associated Cas proteins, and CARD facilitates the detection of antimicrobial resistance genes. Through this comprehensive approach, a thorough understanding of the genomic landscape of \\u003cem\\u003eL. pentosus\\u003c/em\\u003e is achieved, enabling insights into its genetic makeup and functional potential\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec32\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eBioinformatic Analysis\\u003c/h2\\u003e \\u003cp\\u003eThe probiotic characteristics of Lactiplantibacillus pentosus were meticulously investigated through blast analysis and manual examination of various proteins annotated by Prokka. This comprehensive investigation unveiled a spectrum of proteins implicated in stress response, adhesion, aggregation, resistance, and immunomodulation. Detailed information regarding these proteins and their NCBI accession numbers are provided in Table\\u0026nbsp;\\u003cspan refid=\\\"Tab9\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003e.\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab3\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 3\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eRepresents the details about the proteins present in the isolated \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strain genome that are involved in stress, adhesion, aggregation, and resistance with NCBI accession number.\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"4\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eSl No.\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eFactors\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eProteins\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eAccession No.\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e1.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eStress\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"2\\\" rowspan=\\\"3\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\" morerows=\\\"2\\\" rowspan=\\\"3\\\"\\u003e \\u003cp\\u003ea. Temperature\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003ecold-shock protein\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMPQ17990.1, MPQ19628.1, MPQ20388.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003echaperonin GroEL\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMPQ20734.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eco-chaperone GroES\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMPQ20733.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eHsp20 family protein\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMPQ17905.1, MPQ18860.1, MPQ19037.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eHsp33 family molecular chaperone HslO\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMPQ20807.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eb. pH\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003ealkaline shock response membrane anchor protein AmaP\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMPQ19576.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"3\\\" rowspan=\\\"4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\" morerows=\\\"3\\\" rowspan=\\\"4\\\"\\u003e \\u003cp\\u003ec. Other Stress\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eUniversal stress protein\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMPQ18033.1, MPQ18120.1, MPQ18171.1, MPQ18841.1, MPQ19344.1, MPQ19437.1, MPQ20001.1, MPQ20233.1, MPQ20274.1, MPQ20395.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003ePeroxide stress protein YaaA\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMPQ17946.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eGlsB/YeaQ/YmgE family stress response membrane protein\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMPQ18166.1, MPQ19575.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eAsp23/Gls24 family envelope stress response protein\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMPQ18230.1, MPQ18256.1, MPQ19578.1, MPQ19579.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e2.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eAdhesion/Aggregation\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"4\\\" rowspan=\\\"5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\" morerows=\\\"4\\\" rowspan=\\\"5\\\"\\u003e \\u003cp\\u003eAdhesion to mucus/epithelial cells/ECM proteins/plasma components/Aggregation\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eMucus-binding protein\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMPQ18214.1, MPQ20578.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eChaperonin GroEL\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMPQ20734.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eElongation factor Tu\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMPQ19535.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eSortase\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMPQ19087.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eGlyceraldehyde-3-phosphate dehydrogenase\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMPQ19912.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e3.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eResistance\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003ebleomycin resistance protein\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMPQ17822.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003esmall multidrug resistance protein\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMPQ17839.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003ecopper resistance protein CopZ\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eMPQ18979.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003eMoreover, the genome-wide exploration for bacteriocin-encoding genes was conducted using BAGEL 4, revealing the presence of diverse bacteriocin types, including pediocin-like bacteriocin (class IIa), plantaricin E, F (class IIb), and a putative bacteriocin (Bovicin 255 variant). Pediocin showed similarity to bacteriocin from \\u003cem\\u003ePediococcus acidilactici.\\u003c/em\\u003e The Plantaricin E and F cluster, a two-peptide bacteriocin with immunity genes, ABC transporter genes, and accessory genes, exhibits similarity to bacteriocin from \\u003cem\\u003eLactiplantibacillus plantarum.\\u003c/em\\u003e Additionally, lactococcin is present within the same node. The putative bacteriocin (Bovicin 255 variant) shares similarities with the bacteriocin from \\u003cem\\u003eStreptococcus mutans\\u003c/em\\u003e UA159. Notably, NCBI protein blast analysis affirmed the existence of garvicin Q family Class II bacteriocin and lactococcin (class IIc), demonstrating the capability of \\u003cem\\u003eL. pentosus\\u003c/em\\u003e to produce a repertoire of Class II bacteriocins for defensive purposes. Figure\\u0026nbsp;\\u003cspan refid=\\\"Fig5\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003eA illustrates the outcomes obtained from BAGEL 4.\\u003c/p\\u003e \\u003cp\\u003eSimilarly, other strains of \\u003cem\\u003eL. pentosus\\u003c/em\\u003e exhibit distinct bacteriocin production profiles. For instance, L. pentosus ZFM94 synthesizes a bacteriocin termed Pentocin, while \\u003cem\\u003eL. pentosus\\u003c/em\\u003e 124-2 produces two bacteriocins with molecular weights of 26.69 kDa and 17.15 kDa [\\u003cspan citationid=\\\"CR82\\\" class=\\\"CitationRef\\\"\\u003e82\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR83\\\" class=\\\"CitationRef\\\"\\u003e83\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eAdditionally, antiSMASH software was employed to conduct an in-depth analysis of secondary metabolite production, revealing the presence of two regions harboring secondary metabolites. Notably, region 6 encompasses a RiPP-like cluster (other unspecified ribosomally synthesized and post-translationally modified peptide product) containing a class II bacteriocin region with an ABC transporter and accessory protein, whereas region 32.1 harbors a T3PKS (Type III polyketide synthase) cluster with a hydroxymethylglutaryl-CoA synthase region. A cluster blast analysis of the bacteriocins cluster showed significant similarity to bacteriocin clusters of different \\u003cem\\u003eL. plantarum\\u003c/em\\u003e strains, with identities ranging from 50 to 58% (Supplementary Figure \\u003cspan refid=\\\"MOESM1\\\" class=\\\"InternalRef\\\"\\u003eS1\\u003c/span\\u003e depicts the results of secondary metabolite search, and Figure S2 depicts the cluster blast results by antiSMASH). This describes the evolutionary relatedness in bacteriocin production. A similar cluster blast was used in \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strain ZFM94 [\\u003cspan citationid=\\\"CR84\\\" class=\\\"CitationRef\\\"\\u003e84\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eThe Plasmid Finder tool identified the presence of a plasmid in the genome with high identity to rep 28 from \\u003cem\\u003eL. plantarum (\\u003c/em\\u003eNCBI accession number CP005948). This finding aligns with earlier reports indicating the presence of plasmids in certain \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strains [\\u003cspan citationid=\\\"CR85\\\" class=\\\"CitationRef\\\"\\u003e85\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eThe CRISPR Cas finder detected five sequences with CRISPR genes, indicating an adaptive immunity mechanism against foreign mobile genetic elements [\\u003cspan citationid=\\\"CR86\\\" class=\\\"CitationRef\\\"\\u003e86\\u003c/span\\u003e]. Nodes 1, 2, 7, 19, and 28 have CRISPR sequences. The genome has CAS-Type IIA and CAS-Type IE. Additionally, prophage regions were predicted by the PHASTER server, with nodes 13 and 15 containing complete and incomplete phage sequences, respectively (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig5\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003eB). These findings are consistent with previous publications, suggesting that the genome of \\u003cem\\u003eL. pentosus\\u003c/em\\u003e MP-10 included CRISPR Cas genes and prophage regions [\\u003cspan citationid=\\\"CR87\\\" class=\\\"CitationRef\\\"\\u003e87\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eFurthermore, CARD analysis revealed the presence of the Van Y glycopeptide resistance gene cluster by strict hits with 29.63% identity (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig5\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003eC). Loose hits (bit score below 500) showed the presence of 203 antibiotic resistance genes with different resistance mechanisms, including antibiotic efflux, antibiotic target protection, antibiotic inactivation, antibiotic target alteration, reduced permeability to antibiotics, and antibiotic target replacement. Resistance drug classes include glycopeptide antibiotics, peptide antibiotics, macrolide antibiotics, lincosamide antibiotics, cephalosporin, tetracycline antibiotics, aminoglycoside antibiotics, fluoroquinolone antibiotics, disinfecting agents, antiseptics, mupirocin-like antibiotics, rifamycin antibiotics, etc. These findings showed that the strain did not have rigorous antibiotic resistance.\\u003c/p\\u003e \\u003cp\\u003eThese comprehensive analyses collectively enhance our understanding of the probiotic traits, bacteriocin production, genomic architecture, and antibiotic resistance profile of \\u003cem\\u003eL. pentosus\\u003c/em\\u003e, underscoring its potential for therapeutic applications and biotechnological exploitation.\\u003c/p\\u003e \\u003cdiv id=\\\"Sec33\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003eProbiotic Characterization\\u003c/h2\\u003e \\u003cp\\u003eThe \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strain underwent assessment for tolerance to bile and acid, revealing its capacity to withstand 0.15% bile salt concentration and acidic conditions with a pH of 3. The strain\\u0026rsquo;s tolerance dynamics were investigated over time intervals of 0, 3, 6, 9, 12, and 24 h, with survivability percentages calculated from corresponding growth values, as depicted in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig6\\\" class=\\\"InternalRef\\\"\\u003e6\\u003c/span\\u003eA, B. Concurrently, cell viability was evaluated at these time points, and viable counts were recorded, as presented in Supplementary Table S2. Notably, the growth and cell viability levels of the Lactiplantibacillus pentosus strain exhibited an increasing trend over time intervals of 0, 3, 6, 9, 12, and 24 h. However, despite this increase, the percentage of survivability, when compared to the control, demonstrated a decreasing trend. This discrepancy suggests that while the bacterial population grows and maintains viability over time, its ability to survive in the presence of bile salt and acidic conditions decreases relative to the control group. This observation indicates that prolonged exposure to bile salt and acidic environments adversely affects the strain\\u0026rsquo;s survivability, as evidenced by a reduction in its relative resilience compared to the control condition. This observation aligns with prior research conducted by Montoro et al., which explored the survivability of various \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strains under analogous conditions [\\u003cspan citationid=\\\"CR80\\\" class=\\\"CitationRef\\\"\\u003e80\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eThe culture of \\u003cem\\u003eL. pentosus\\u003c/em\\u003e was screened for bile salt hydrolase (BSH) activity using a direct plate assay. BSH activity is recognized to facilitate bacterial colonization within the gastrointestinal tract [\\u003cspan citationid=\\\"CR88\\\" class=\\\"CitationRef\\\"\\u003e88\\u003c/span\\u003e]. Notably, the isolated \\u003cem\\u003eL. pentosus strain\\u003c/em\\u003e produced a discernible white precipitate zone in bile agar media surrounding the colony, indicative of BSH production. Bifidobacterium adolescentis was the positive control used to validate this observation, while \\u003cem\\u003eS.\\u003c/em\\u003e Typhi was employed as the negative control (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig7\\\" class=\\\"InternalRef\\\"\\u003e7\\u003c/span\\u003eA). Moreover, it is noteworthy that in earlier research, bile salt hydrolase (BSH) activity has been reported in several Lactiplantibacillus species originating from food and human sources. For instance, it has been observed that strains like \\u003cem\\u003eL. pentosus\\u003c/em\\u003e CHIG have positive BSH activity [\\u003cspan citationid=\\\"CR89\\\" class=\\\"CitationRef\\\"\\u003e89\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eFurthermore, the strain demonstrated 57.32% lysozyme tolerance with a CFU/mL count of 9.5 \\u0026times; 10\\u003csup\\u003e7\\u003c/sup\\u003e after 1 h of incubation. This finding underscores the strain\\u0026rsquo;s capability to withstand lysozyme exposure.\\u003c/p\\u003e \\u003cp\\u003eGastrointestinal tolerance of the isolated \\u003cem\\u003eL. pentosus\\u003c/em\\u003e was assessed over intervals of 0, 3, 6, 9, 12, and 24 h. The results were similar to those observed for acid bile tolerance, with survivability percentages of the isolated strain in gastrointestinal conditions compared to control MRS documented in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig6\\\" class=\\\"InternalRef\\\"\\u003e6\\u003c/span\\u003e. Notably, an inverse relationship was observed between survivability percentage and time duration, wherein an increase in incubation time corresponded to a decrease in survivability percentage. The bacterial population is growing and maintaining viability over time; however, its ability to survive in the presence of gastrointestinal conditions decreases relative to the control group. Furthermore, Supplementary Table S2 provides a comprehensive overview of viable counts obtained at each time point, further elucidating the strain\\u0026rsquo;s dynamics in gastrointestinal conditions. These findings underscore the strain\\u0026rsquo;s ability to endure the challenges presented by the gastrointestinal environment. Such insights are pivotal for assessing the strain\\u0026rsquo;s potential as a probiotic agent and its suitability for gastrointestinal health applications.\\u003c/p\\u003e \\u003cp\\u003eTo evaluate its adhesion capability, the strain underwent testing for cell surface hydrophobicity using hydrocarbons, namely xylene, chloroform, and n-hexadecane. The observed hydrophobicity levels were 21.12\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.94% in the presence of xylene, 13.53\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.09% in the presence of n-hexadecane, and 26.72\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.86% in the presence of chloroform. Comparable hydrophobicity profiles have been reported in other strains, such as \\u003cem\\u003eLactiplantibacillus pentosus\\u003c/em\\u003e isolated from fermented fish, \\u003cem\\u003eLactococcus lactis\\u003c/em\\u003e, and \\u003cem\\u003eLactobacillus fermentum\\u003c/em\\u003e [\\u003cspan citationid=\\\"CR90\\\" class=\\\"CitationRef\\\"\\u003e90\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eProbiotic species\\u0026rsquo; radical scavenging capacity is an important characteristic, indicating their antioxidant nature. This was assessed using the DPPH radical scavenging assay, where the isolated \\u003cem\\u003eL. pentosus\\u003c/em\\u003e demonstrated 60.70% DPPH activity, showcasing its antioxidative potential. Control ascorbic acid (10 \\u0026micro;g/mL) gave 42.75% DPPH activity. Notably, studies by Unban et al. reported even higher DPPH scavenging activity in \\u003cem\\u003eL. pentosus\\u003c/em\\u003e A14-6 and \\u003cem\\u003eL. pentosus\\u003c/em\\u003e A26-8 [\\u003cspan citationid=\\\"CR91\\\" class=\\\"CitationRef\\\"\\u003e91\\u003c/span\\u003e]. A proteolytic assay was conducted using a skim milk agar plate, with trypsin (1.0 mg/mL) as the positive control, resulting in 19 mm of proteolysis. However, the supernatant and pellet of the isolated \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strain did not exhibit any zone of inhibition in the test, indicating the absence of proteolytic activities.\\u003c/p\\u003e \\u003cp\\u003eThe antibiotic susceptibility profile of the \\u003cem\\u003eL. pentosus\\u003c/em\\u003e isolate was assessed against a panel of 20 antibiotics, revealing resistance to cefoxitin (CX), amoxicillin/clavulanic acid (AMC), and amikacin (AK). However, sensitivity or intermediate resistance was observed towards all other antibiotics tested (refer to Table\\u0026nbsp;\\u003cspan refid=\\\"Tab10\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003e and Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig7\\\" class=\\\"InternalRef\\\"\\u003e7\\u003c/span\\u003eB, C), indicating the strain is susceptible to most antibiotics, thereby supporting its safety for probiotic applications [\\u003cspan citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e27\\u003c/span\\u003e]. This finding aligns with observations by Cazodo Munoz et al., who noted that out of 59 \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strains tested, a majority (95%) of strains were resistant to at least 3 antibiotics when evaluated against 15 antibiotics [\\u003cspan citationid=\\\"CR92\\\" class=\\\"CitationRef\\\"\\u003e92\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab4\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 4\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eRepresents antibiotic susceptibility results of the isolated \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strain.\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"5\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eSl No.\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eAntibiotics\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eConcentration\\u003c/p\\u003e \\u003cp\\u003e(\\u0026micro;g/Disc)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eZone of Inhibition\\u003c/p\\u003e \\u003cp\\u003e(in mm)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eResistance/\\u003c/p\\u003e \\u003cp\\u003eSensitive\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eUniversal-1 OD308\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e1.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eGentamicin (GEN)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e10\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e17\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eI\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e2.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eAmikacin (AK)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e12\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eR\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e3.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eCiprofloxacin (CIP)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e5\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e15\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eI\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e4.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eCefoxitin (CX)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eR\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e5.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eAmoxycillin/Clavulanic acid (AMC)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e20/10\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e11\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eR\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e6.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eTetracycline (TE)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e18\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eI\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e7.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eChloramphenicol (C)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eS\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e8.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eCo-trimoxazole (COT)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e16\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eI\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eDodeca Universal-XII DE027\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e9.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eOfloxacin (OF)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e5\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e16\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eI\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e10.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eCefadroxil (CFR)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e21\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eS\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e11.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eDoxycycline HCl (DO)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e16\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eI\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e12.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eCloxacillin (COX)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e5\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e18\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eI\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e13.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eAzithromycin (AZM)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e21\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eS\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e14.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eCefotaxime (CTX)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e10\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e20\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eS\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e15.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eCeftriaxone (CTR)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e20\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eS\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e16.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eTicarcillin (TI)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e75\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e17\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eI\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e17.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003ePiperacillin/ Tazobactam (PIT)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e100/10\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eS\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e18.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eCiprofloxacin (CIP)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e5\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e15\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eI\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e19.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eLevofloxacin (LE)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e5\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e21\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eS\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e20.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eCeftazidime (CAZ)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e22\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eS\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003ctfoot\\u003e \\u003ctr\\u003e\\u003ctd colspan=\\\"5\\\"\\u003eR\\u0026thinsp;=\\u0026thinsp;resistant (\\u0026le;\\u0026thinsp;14 mm); S\\u0026thinsp;=\\u0026thinsp;sensitive (\\u0026ge;\\u0026thinsp;20 mm); I\\u0026thinsp;=\\u0026thinsp;intermediate (15\\u0026ndash;19 mm).\\u003c/td\\u003e\\u003c/tr\\u003e \\u003c/tfoot\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003eA hemolytic assay was conducted for the safety assessment, wherein the cells and cell-free supernatant of the strain did not exhibit detectable hemolytic activity. However, the 10-fold concentrated cell-free supernatant showed media color diffusion (yellow to green color) with no clear zone of inhibition, indicative of potential partial hemolysis (α hemolysis) or absence of hemolysis. Triton x100 was used as the positive control, displaying a clear zone around the well (β hemolysis). In contrast, Nisin (1.0 mg/mL) was the negative control, demonstrating no hemolytic activity (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig7\\\" class=\\\"InternalRef\\\"\\u003e7\\u003c/span\\u003eD). Similar findings were reported for \\u003cem\\u003eL. pentosus\\u003c/em\\u003e 22C, isolated from traditional yogurt, which also lacked hemolytic activity [\\u003cspan citationid=\\\"CR93\\\" class=\\\"CitationRef\\\"\\u003e93\\u003c/span\\u003e]. The absence of clear hemolytic (β) activities confirms that the strain is safe to use.\\u003c/p\\u003e \\u003cp\\u003eFurthermore, the strain exhibited 23% auto-aggregation (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig8\\\" class=\\\"InternalRef\\\"\\u003e8\\u003c/span\\u003eA) and 51% co-aggregation with \\u003cem\\u003eS.\\u003c/em\\u003e Typhi (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig8\\\" class=\\\"InternalRef\\\"\\u003e8\\u003c/span\\u003eB). Previous reports have indicated that \\u003cem\\u003eL. pentosus\\u003c/em\\u003e exhibits notable co-aggregation abilities, including solid co-aggregation with \\u003cem\\u003eStreptococcus mutans\\u003c/em\\u003e [\\u003cspan citationid=\\\"CR94\\\" class=\\\"CitationRef\\\"\\u003e94\\u003c/span\\u003e]. Many \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strains isolated from naturally fermented Alore\\u0026ntilde;a table olives exhibited similar auto-aggregation and co-aggregation. Among them, 19% of \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strains had a high capability to auto-aggregate (50\\u0026ndash;77.92%), while 42% had a medium auto-aggregation capacity (35\\u0026ndash;50%). They also showed different ranges of co-aggregation with pathogens, including \\u003cem\\u003eE. coli\\u003c/em\\u003e, \\u003cem\\u003eSalmonella\\u003c/em\\u003e, \\u003cem\\u003eListeria innocua\\u003c/em\\u003e, and \\u003cem\\u003eStaphylococcus aureus\\u003c/em\\u003e [\\u003cspan citationid=\\\"CR80\\\" class=\\\"CitationRef\\\"\\u003e80\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eMoreover, the antibacterial activity of the cell-free supernatant (CFS) was evaluated using an agar-well diffusion assay, revealing antibacterial activity against various pathogens, including both Gram-positive and Gram-negative strains. Notably, in some instances, the antibacterial activity of the CFS was comparable to streptomycin (antibiotic) and superior to nisin (commercially available bacteriocin). Further details are provided in Table\\u0026nbsp;\\u003cspan refid=\\\"Tab11\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003e. These observations are consistent with findings reported for \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strains provided by CICC (China Centre of Industrial Culture Collection) [\\u003cspan citationid=\\\"CR95\\\" class=\\\"CitationRef\\\"\\u003e95\\u003c/span\\u003e] and \\u003cem\\u003eL. pentosus\\u003c/em\\u003e ZFM94, isolated from infant faces [\\u003cspan citationid=\\\"CR82\\\" class=\\\"CitationRef\\\"\\u003e82\\u003c/span\\u003e]. These evaluations collectively offer insights into various functional characteristics of \\u003cem\\u003eL. pentosus\\u003c/em\\u003e, contributing to a comprehensive understanding of its probiotic attributes.\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab5\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 5\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eRepresents the antibacterial activity of CFS of the isolated \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strain against different food pathogens.\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"5\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eSl No.\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eBacteria\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eZOI\\u003c/p\\u003e \\u003cp\\u003eCell-Free Supernatant in mm\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eZOI Nisin\\u003c/p\\u003e \\u003cp\\u003ein mm\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eZOI Streptomycin in mm\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e1.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eListeria monocytogenes\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e19\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e10\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e19\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e2.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eVibrio harveyi\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e16\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e14\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e13\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e3.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eStreptococcus mutans\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e16\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e13\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e13\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e4.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eStaphylococcus aureus\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e16\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e10\\u003c/p\\u003e \\u003cp\\u003e(not clear)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e16\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e5.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eStreptococcus thermophilus\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e18\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e14\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e6.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eCorynebacterium callunae\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e20\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e11\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e20\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e7.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eEnterococcus gallinarum\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e18\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e14\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e19\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e8.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eVibrio cholerae\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e16\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e14\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e9.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eBacillus cereus\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e18\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e13\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e10.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003ePseudomonas putida\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e16\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e19\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e11.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eEscherichia coli\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e16\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e20\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e12.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003ePseudomonas aeruginosa\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e16\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e10\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e16\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e13.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eClostridium perfringens\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e21\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e16\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e14.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eSalmonella enterica subsp. enterica\\u003c/em\\u003e ser. Typhi\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e18\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e16\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e15.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eSalmonella enterica subsp. enterica\\u003c/em\\u003e ser. Paratyphi\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e17\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e17\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003ctfoot\\u003e \\u003ctr\\u003e\\u003ctd colspan=\\\"5\\\"\\u003eZOI is the zone of inhibition, and CFS is the cell-free supernatant of the isolated \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strain.\\u003c/td\\u003e\\u003c/tr\\u003e \\u003c/tfoot\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eTable 6:\\u003c/strong\\u003e Antibacterial activity against various food pathogens. ZOI= Zone of Inhibition, CFS= Cell free supernatant.\\u003c/p\\u003e\\n\\u003ctable border=\\\"1\\\" cellspacing=\\\"0\\\" cellpadding=\\\"0\\\" width=\\\"499\\\"\\u003e\\n \\u003ctbody\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"9.839357429718875%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eSl No.\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"27.91164658634538%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eBacteria\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.281124497991968%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eZOI CFS\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003ein mm\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"21.08433734939759%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eZOI Nisin\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003ein mm\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.883534136546185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eZOI Streptomycin in mm\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"9.839357429718875%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"27.91164658634538%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eListeria monocytogenes\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.281124497991968%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e19\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"21.08433734939759%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e10\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.883534136546185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e19\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"9.839357429718875%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"27.91164658634538%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eVibrio harveyi\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.281124497991968%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e16\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"21.08433734939759%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e14\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.883534136546185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e13\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"9.839357429718875%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e3\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"27.91164658634538%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eStreptococcus mutans\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.281124497991968%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e16\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"21.08433734939759%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e13\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.883534136546185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e13\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"9.839357429718875%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e4\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"27.91164658634538%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eStaphylococcus aureus\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.281124497991968%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e16\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"21.08433734939759%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e10\\u003c/p\\u003e\\n \\u003cp\\u003e(not clear)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.883534136546185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e16\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"9.839357429718875%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e5\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"27.91164658634538%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eStreptococcus thermophilus\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.281124497991968%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e18\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"21.08433734939759%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.883534136546185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e14\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"9.839357429718875%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e6\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"27.91164658634538%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eCorynebacterium callunae\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.281124497991968%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e20\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"21.08433734939759%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e11\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.883534136546185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e20\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"9.839357429718875%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e7\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"27.91164658634538%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eEnterococcus gallinarum\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.281124497991968%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e18\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"21.08433734939759%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e14\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.883534136546185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e19\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"9.839357429718875%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e8\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"27.91164658634538%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eVibrio cholerae\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.281124497991968%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e16\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"21.08433734939759%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.883534136546185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e14\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"9.839357429718875%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e9\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"27.91164658634538%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eBacillus cereus\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.281124497991968%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e18\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"21.08433734939759%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e13\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.883534136546185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e25\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"9.839357429718875%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e10\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"27.91164658634538%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003ePseudomonas putida\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.281124497991968%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e16\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"21.08433734939759%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.883534136546185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e19\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"9.839357429718875%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e11\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"27.91164658634538%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eEscherichia coli\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.281124497991968%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e16\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"21.08433734939759%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.883534136546185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e20\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"9.839357429718875%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e12\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"27.91164658634538%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003ePseudomonas aeruginosa\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.281124497991968%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e16\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"21.08433734939759%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e10\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.883534136546185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e16\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"9.839357429718875%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e13\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"27.91164658634538%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eClostridium perfringens\\u0026nbsp;\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.281124497991968%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e21\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"21.08433734939759%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.883534136546185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e16\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"9.839357429718875%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e14\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"27.91164658634538%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eSalmonella typhi\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.281124497991968%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e18\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"21.08433734939759%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.883534136546185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e16\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"9.839357429718875%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e15\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"27.91164658634538%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eSalmonella paratyphi\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.281124497991968%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e17\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"21.08433734939759%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"20.883534136546185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e17\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003c/tbody\\u003e\\n\\u003c/table\\u003e\"},{\"header\":\"Conclusions\",\"content\":\"\\u003cp\\u003eThe genomic and probiotic characterization of the \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strain isolated from fermented rice revealed that the strain is a potential probiotic that can be used in the food industry. The genome study revealed that it carries genes that play a significant role in adhesion, aggregation, stress tolerance, antibiotic resistance, CRISPR CAS, and secondary metabolites, including bacteriocin. It also showed its phylogenetic relationship with \\u003cem\\u003eL. plantarum\\u003c/em\\u003e. Probiotic characterization through different assays revealed the growth in various extreme situations, including low pH, the presence of bile salt, intestinal and gastric conditions, and their antioxidant potential. Additionally, the non-hemolysis and non-proteolytic natures were also demonstrated. Its broad range of antibacterial activity, as well as its antibiotic sensitivity and resistance, was also disclosed. These comprehensive findings collectively underscore the suitability of this strain for probiotic applications within the food industry, implying that it might be a promising prospect for functional food development and human health enhancement initiatives.\\u003c/p\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003eSupplementary Materials:\\u003c/strong\\u003e Figure S1: Illustrates the identification of biosynthetic gene clusters by antiSMASH 6.0 from the isolated\\u003cem\\u003e\\u0026nbsp;L. pentosus\\u003c/em\\u003e genome. The predicted gene cluster showed a significant hit with RiPP-like and T3PKS clusters: a. Region 6.1 represents the RiPP-like cluster, and other unspecified ribosomally synthesized and post-translationally modified peptide products consist of two peptide bacteriocin clusters with ABC transporter and accessory gene; b. Region 32.1-T3PKS-Type III Polyketide synthase cluster with hydroxymethylglutaryl-CoA synthase region. Figure S2: Cluster Blast of RiPP region of \\u003cem\\u003eL. pentosus\\u003c/em\\u003e in anti-SMASH shows the different genomes with similar bacteriocin clusters and their percentage of identity. Table S1: Sugar utilization pattern of isolated \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strain. Table S2: Represents the viability of the isolated \\u003cem\\u003eL. pentosus\\u0026nbsp;\\u003c/em\\u003estrain in the presence of acid, bile salt, and gastric-intestinal conditions.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAuthor Contributions:\\u003c/strong\\u003e Conceptualization: R.K. and AC.; methodology: A.C.; writing\\u0026mdash;original draft preparation: A.C.; funding acquisition: R.K. All authors have read and agreed to the published version of the manuscript.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eFunding:\\u003c/strong\\u003e This work was supported by the LSRB (The Life Sciences Research Board (LSRB) of Defense Research \\u0026amp; Development Organization, GAP-537 and MOFPI (Ministry of Food Processing Industries, GAP-536). CSIR provides funding for researchers. We thank the Director, CSIR-CFTRI, Mysore, for providing the facilities.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eData Availability Statement:\\u003c/strong\\u003e The original contributions presented in the study are included in the article/Supplementary Material, further inquiries can be directed to the corresponding author.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConflicts of Interest:\\u003c/strong\\u003e The authors declare no conflicts of interest.\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\n\\u003cli\\u003eNarzary Y, Brahma J, Brahma C et al (2016) A study on indigenous fermented foods and beverages of Kokrajhar, Assam, India. J. Ethn. Foods 3: 284\\u0026ndash;291. https://doi.org/10.1016/j.jef.2016.11.010.\\u003c/li\\u003e\\n\\u003cli\\u003eRezac S, Kok CR, Heermann M et al (2018) Fermented foods as a dietary source of live organisms. Front. 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Charact. 11: 972\\u0026ndash;978. https://doi.org/10.1007/s11694-017-9471-z.\\u003c/li\\u003e\\n\\u003cli\\u003eGu M, Cho JH, Suh JW et al (2023) Potential oral probiotic \\u003cem\\u003eLactobacillus pentosus\\u003c/em\\u003e MJM60383 inhibits \\u003cem\\u003eStreptococcus mutans\\u003c/em\\u003e biofilm formation by inhibiting sucrose decomposition. J. Oral Microbiol. 15: 2161179. https://doi.org/10.1080/20002297.2022.2161179.\\u003c/li\\u003e\\n\\u003cli\\u003eZhu Y, Zhang S (2020) Antibacterial activity and mechanism of lacidophilin from Lactobacillus pentosus against \\u003cem\\u003eStaphylococcus aureus\\u003c/em\\u003e and \\u003cem\\u003eEscherichia Coli.\\u003c/em\\u003e Front. Microbiol. 11: 582349. https://doi.org/10.3389/fmicb.2020.582349.\\u003c/li\\u003e\\n\\u003c/ol\\u003e\"}],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":true,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":false,\"hideJournal\":false,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":true,\"isAuthorSuppliedPdf\":false,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":false,\"isPdf\":false,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"probiotics-and-antimicrobial-proteins\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"paap\",\"sideBox\":\"Learn more about [Probiotics and Antimicrobial Proteins](http://link.springer.com/journal/12601)\",\"snPcode\":\"12602\",\"submissionUrl\":\"https://submission.nature.com/new-submission/12602/3\",\"title\":\"Probiotics and Antimicrobial Proteins\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"em\",\"reportingPortfolio\":\"Springer Hybrid\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false},\"keywords\":\"L. pentosus, whole genome sequence, bacteriocin, gastrointestinal tract, antimicrobial\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-4845642/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-4845642/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003cp\\u003eThis study focused on a potential probiotic strain of \\u003cem\\u003eLactiplantibacillus pentosus\\u003c/em\\u003e (strain krglsrbmofpi2) isolated from traditional fermented rice in India. The study aimed to conduct preliminary genetic and phenotypic characterization. The strain has a genome size of 3.7 Mb, a GC content of 46%, and 3192 protein-coding sequences, as determined by genomic analysis. Subsequently, various bacteriocins, the CRISPR Cas system, phage genes, plasmids, pathogenicity, and antibiotic resistance were identified using bioinformatic methodologies. Biochemical and biophysical examinations improved our understanding of hydrophobicity, antioxidant activity, antibiotic resistance, auto-aggregation, co-aggregation properties, and tolerance to the simulated gastrointestinal condition. The safety assessment of the isolated \\u003cem\\u003eL. pentosus\\u003c/em\\u003e was evaluated based on the hemolytic activity. The strain has demonstrated strong antibiotic activity against pathogens considered WHO priorities, such as \\u003cem\\u003eSalmonella enterica\\u003c/em\\u003e subsp. \\u003cem\\u003eenterica\\u003c/em\\u003e ser. Typhi, \\u003cem\\u003eClostridium perfringens\\u003c/em\\u003e, \\u003cem\\u003eEscherichia coli\\u003c/em\\u003e, \\u003cem\\u003eListeria monocytogenes\\u003c/em\\u003e, \\u003cem\\u003eStaphylococcus aureus\\u003c/em\\u003e, and \\u003cem\\u003eVibrio cholerae\\u003c/em\\u003e. Our findings suggest that gaining a deep understanding of the genetic and functional characteristics of the \\u003cem\\u003eL. pentosus\\u003c/em\\u003e strain could pave the way for its application as a beneficial probiotic in the food industry.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Genomic Characterization and Probiotic Properties of Lactiplantibacillus pentosus Isolated from Fermented Rice\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2024-09-03 05:01:23\",\"doi\":\"10.21203/rs.3.rs-4845642/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0},{\"type\":\"decision\",\"content\":\"Revision requested\",\"date\":\"2024-09-02T13:59:18+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorInvitedReview\",\"content\":\"\",\"date\":\"2024-09-01T06:59:36+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"134042173444364065941167844757970957188\",\"date\":\"2024-08-29T04:27:17+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"editorInvitedReview\",\"content\":\"\",\"date\":\"2024-08-28T17:48:43+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"119887234616270664453825171327240332895\",\"date\":\"2024-08-28T05:45:33+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"302769184358606479280418580903941987763\",\"date\":\"2024-08-25T05:14:15+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"331179896599549389203650573941029667643\",\"date\":\"2024-08-23T07:58:29+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"272218061229077257435612663208644526799\",\"date\":\"2024-08-23T03:31:56+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"205526857843596911902800461345739459472\",\"date\":\"2024-08-23T02:37:41+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewersInvited\",\"content\":\"\",\"date\":\"2024-08-22T21:08:43+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorAssigned\",\"content\":\"\",\"date\":\"2024-08-08T05:52:31+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"checksComplete\",\"content\":\"\",\"date\":\"2024-08-08T05:50:58+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"submitted\",\"content\":\"Probiotics and Antimicrobial Proteins\",\"date\":\"2024-08-02T04:51:05+00:00\",\"index\":\"\",\"fulltext\":\"\"}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"probiotics-and-antimicrobial-proteins\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"paap\",\"sideBox\":\"Learn more about [Probiotics and Antimicrobial Proteins](http://link.springer.com/journal/12601)\",\"snPcode\":\"12602\",\"submissionUrl\":\"https://submission.nature.com/new-submission/12602/3\",\"title\":\"Probiotics and Antimicrobial Proteins\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"em\",\"reportingPortfolio\":\"Springer Hybrid\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false}}],\"origin\":\"\",\"ownerIdentity\":\"44ef3728-92bb-4a9b-8db3-e63633513a69\",\"owner\":[],\"postedDate\":\"September 3rd, 2024\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"published-in-journal\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2024-10-28T16:05:28+00:00\",\"versionOfRecord\":{\"articleIdentity\":\"rs-4845642\",\"link\":\"https://doi.org/10.1007/s12602-024-10378-1\",\"journal\":{\"identity\":\"probiotics-and-antimicrobial-proteins\",\"isVorOnly\":false,\"title\":\"Probiotics and Antimicrobial Proteins\"},\"publishedOn\":\"2024-10-21 15:58:09\",\"publishedOnDateReadable\":\"October 21st, 2024\"},\"versionCreatedAt\":\"2024-09-03 05:01:23\",\"video\":\"\",\"vorDoi\":\"10.1007/s12602-024-10378-1\",\"vorDoiUrl\":\"https://doi.org/10.1007/s12602-024-10378-1\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-4845642\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-4845642\",\"identity\":\"rs-4845642\",\"version\":[\"v1\"]},\"buildId\":\"-HB7Z8yhvgn0wM9Nzuekk\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}