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Xylanase is an essential industrial enzyme for degrading plant biomass, pulp and paper, textiles, bio-scouring, food, animal feed, biorefinery, chemicals, and pharmaceutical industries. Despite its significant industrial importance, the extensive application of xylanase is hampered by high production costs and concerns regarding the safety of xylanase-producing microorganisms. The utilisation of renewable polymers for enzyme production is becoming a cost-effective alternative. Among the prospective candidates, non-pathogenic lactic acid bacteria (LAB) are promising for safe and eco-friendly applications. Our investigation revealed that Pediococcus pentosaceus G4, isolated from plant sources, is a notable producer of extracellular xylanase. Improving the production of extracellular xylanase is crucial for viable industrial applications. Therefore, the current study investigated the impact of various medium components and optimised the selected medium composition for extracellular xylanase production of P. pentosaceus G4 using Plackett–Burman Design (PBD) and Central Composite Design (CCD) statistical approaches. According to BPD analysis, 8 out of the 19 investigated factors (glucose, almond shell, peanut shell, walnut shell, malt extract, xylan, urea, and magnesium sulphate) demonstrated significant positive effects on extracellular xylanase production of P. pentosaceus G4. Among them, glucose, almond shells, peanut shells, urea, and magnesium sulphate were identified as the main medium components that significantly (p < 0.05) influenced the production of extracellular xylanase of P. pentosaceus G4. The optimal concentrations of glucose, almond shells, peanut shells, urea, and magnesium sulphate, as determined via CCD, were 26.87 g/L, 16 g/L, 30 g/L, 2.85 g/L, and 0.10 g/L, respectively. The optimised concentrations resulted in extracellular xylanase activity of 2.765 U/mg, which was similar to the predicted extracellular xylanase activity of 2.737 U/mg. The CCD-optimised medium yielded a 3.13-fold enhancement in specific extracellular xylanase activity and a 7.99-fold decrease in production costs compared to the commercial de Man, Rogosa and Sharpe medium, implying that the CCD-optimised medium is a cost-effective medium for extracellular xylanase production of P. pentosaceus G4. Moreover, this study demonstrated a positive correlation between extracellular xylanase production, growth, lactic acid production and the amount of sugar utilised, implying the multifaceted interactions of the physiological variables affecting extracellular xylanase production in P. pentosaceus G4. In conclusion, statistical methods are effective in rapidly assessing and optimising the medium composition to enhance extracellular xylanase production of P. pentosaceus G4. Furthermore, the findings of this study highlighted the potential of using LAB as a cost-effective producer of extracellular xylanase enzymes using optimised renewable polymers, offering insights into the future use of LAB in producing hemicellulolytic enzymes.

Currently, there is an increasing use of whole‐genome sequencing (WGS) studies to investigate the molecular taxonomy, metabolic properties, enzyme capabilities, and bioactive substances of lactic acid bacteria (LAB) species. In this study, the genome of strain Pediococcus pentosaceus BBS1 was sequenced using the Illumina HiSeq. 2500 platform to determine its classification, annotate its main features, and evaluate its safety characteristics. Results showed an average nucleotide identity (ANI) value of 99.60% for Pediococcus pentosaceus BBS1. P. pentosaceus BBS1 genome was composed of a 1,840,613 bp circular chromosome with a GC content of 37.23%, which contained 1778 predicted protein‐coding sequences (CDSs). Rapid Annotation using Subsystems Technology (RAST) linked to the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that strain BBS1 possesses l‐acetate dehydrogenase (l‐LDH; EC 1.1.1.27) and d‐lactate dehydrogenase (d‐LDH; EC 1.1.1.28), which are the genes responsible for lactic acid production. Additionally, it was found to contain linamarase, or β‐glucosidases (EC 3.2.1.21), a gene that functions for cyanide degradation. Significantly, the safety studies carried out using WGS confirmed the absence of virulence factors, biogenic amines, and antibiotic‐resistance genes in BBS1. Our previous research conducted in this study have shown that BBS1 possesses probiotic features, including tolerance to the simulated artificial gastrointestinal tract, bacterial adhesion, antibacterial activity, and antioxidant function. The findings provided herein significantly enhanced the known information on BBS1, supporting its potential application in promoting health through food products. Pediococcus pentosaceus BBS1 was analyzed by whole genome sequencing for its classification, annotated its main features for significant genes, and safety characteristics. The strain has the potential to be used as a beneficial probiotic starter culture, improving the quality of the fermented bamboo and related products.

The development of non-antibiotic and environmentally friendly agents is a key consideration for health management in shrimp aquaculture. In this study, the probiotic potential in shrimp aquaculture of Pediococcus pentosaceus MR001, isolated from Macrobrachium rosenbergii , was investigated by means of feeding trial and genetic characterization. In the feeding trial, dietary supplementation with P. pentosaceus MR001 significantly increased weight gain and digestive enzyme activity ( p  < 0.05) in shrimp, Litopenaeus vannamei . The intestinal histology showed that shrimp given the probiotic diet had healthier guts than the control group. Also, the immune gene expression and the survival rate in the treatment group were significantly increased when compared with the control group. The genetic characteristics of P. pentosaceus strain MR001 were explored by performing whole-genome sequencing (WGS) using the HiSeq 2500 platform and PacBio system, revealing the complete circular genome of 1,804,896 bp. We also identified 1789 coding genes and subsequently characterized genes related to the biosynthesis of bacteriocins, stress resistance, and bile tolerance. Our findings suggest that insights in the functional and genetic characteristics of P. pentosaceus strain MR001 could provide opportunities for applications of such strain in shrimp diet supplementation.

Bacteriocins are ribosomal-synthesized peptides with antimicrobial activity, produced by different groups of bacteria, including lactic acid bacteria (LAB). Most of the produced by LAB bacteriocins can be described with rather broad spectra of inhibition and they offer suggested applications in food preservation and pharmaceutical sector. Different LAB were isolated from fermented food products and fruits, obtained from the region of Pohang, Korea, and identified based on physiological, biochemical, and molecular methods. The promising isolates, Pediococcus pentosaceus 732, Lactococcus lactis 808, and Lactococcus lactis subsp. lactis 431, were identified based on biochemical, physiological, and biomolecular approaches, including 16S rRNA partial sequencing, and were evaluated for production of bacteriocin, including stability in presence of enzymes, chemicals, pH, and temperatures. Adherence properties for the expressed bacteriocins by P. pentosaceus 732, Lc. lactis 808, and Lc. lactis subsp. lactis 431 were evaluated at presence of selected chemicals, pH, and temperatures. The presence of bacteriocin genes in the strains was investigated and analyzed. The bacterial effect of bacteriocin produced by studied strains on Listeria spp. and Staphylococcus spp. has been shown for actively growing and stationary cells. Similar growth and bacteriocin production were observed when studied strains were cultured in MRS at 30 °C or 37 °C. The presence of nisin operon with some point mutations on the genomic DNA was recorded based on the performed PCR reactions targeting different genes associated with nisin expression for both lactococcal strains. Pediocin PA-1 operon was evaluated in a similar manner for P. pentosaceus 732.

Pediococcus pentosaceus is well known for its probiotic properties, including roles in improving health, antimicrobial production, and enhancing fermented food quality. This study aimed to comprehensively analyze the whole genome of P . pentosaceus MBBL6, isolated from healthy cow milk, to assess its probiotic and antimicrobial potentials. P . pentosaceus MBBL6, isolated from a healthy cow milk at BSMRAU dairy farm, Gazipur, Bangladesh, underwent comprehensive genomic analysis, including whole genome sequencing, assembly, annotation, phylogenetic comparison, and assessment of metabolic pathways and secondary metabolites. Antimicrobial efficacy was evaluated through in-vitro and in-vivo studies, alongside in-silico exploration for potential mastitis therapy. We predicted 1,906 genes and 204 SEED sub-systems involved in carbohydrate metabolism and vitamin B complex biosynthesis, with a focus on lactose metabolism in MMBL6. Notably, 43 putative carbohydrate-active enzyme genes, including lysozymes, suggest the ability of MBBL6 for carbohydrate biotransformation and antimicrobial activity. The genome also revealed primary metabolic pathways for arginine and gallic acid metabolism and secondary metabolite gene clusters, including T3PKS and RiPP-like regions. Importantly, two bacteriocin biosynthesis gene clusters namely bovicin_255 _variant and penocin_A , were identified in MBBL6. The safety assessment of MBBL6 genome revealed no virulence genes and a low pathogenicity score (0.196 out of 1.0). Several genes related to survival in gastrointestinal tract and colonization were also identified. Furthermore, MBBL6 exhibited susceptibility to a wide range of antibiotics in-vitro , and effectively suppressed mastitis pathogens in an in-vivo mouse mastitis model trial. The observed bacteriocin, particularly bovicin, demonstrated the ability to disrupt the function of an essential protein, Rho factor of mastitis pathogens by blocking transcription termination process. Taken together, our in-depth genomic analysis underscores the metabolic versatility, safety profile, and antimicrobial potential of P . pentosaceus MBBL6, suggesting its promise for applications in therapeutics, bioremediation, and biopreservation.

Background The genomic information available for Pediococcus pentosaceus is primarily derived from fermented fruits and vegetables, with less information available from fermented meat. P. pentosaceus LL-07, a strain isolated from fermented meat, has the capability of producing exopolysaccharides (EPS). To assess the probiotic attributes of P. pentosaceus LL-07, we conducted whole-genome sequencing (WGS) using the PacBio SequelIIe and Illumina MiSeq platforms, followed by in vitro experiments to explore its probiotic potential. Results The genome size of P. pentosaceus LL-07 is 1,782,685 bp, comprising a circular chromosome and a circular plasmid. Our investigation revealed the absence of a CRISPR/Cas system. Sugar fermentation experiments demonstrated the characteristics of carbohydrate metabolism. P. pentosaceus LL-07 contains an EPS synthesis gene cluster consisting of 13 genes, which is different from the currently known gene cluster structure. NO genes associated with hemolysis or toxin synthesis were detected. Additionally, eighty-six genes related to antibiotic resistance were identified but not present in the prophage, transposon or plasmid. In vitro experiments demonstrated that P. pentosaceus LL-07 was comparable to the reference strain P. pentosaceus ATCC25745 in terms of tolerance to artificial digestive juice and bile, autoaggregation and antioxidation, and provided corresponding genomic evidence. Conclusion This study confirmed the safety and probiotic properties of P. pentosaceus LL-07 via complete genome and phenotype analysis, supporting its characterization as a potential probiotic candidate.

Pediococcus pentosaceus Li05, a strain of lactic acid bacteria isolated from the faeces of healthy volunteers, exhibited potential protective effects against various diseases. This study performed third‐generation sequencing and detailed characterisation of its genome. The Li05 chromosome harboured conserved genes associated with acid resistance (atp), bile salt resistance (bsh), oxidative stress resistance (hsl, dltA, and et al.), and adhesion (nrd, gap, and et al.), whereas the plasmid did not contain antibiotic resistance or virulence genes. Following intervention with Li05 in loperamide‐induced constipated mice, constipation symptoms improved. Meanwhile, alterations in gut microbiota, increased BSH activity in faeces, and modifications to the faecal bile acid profile were observed. Additionally, expression levels of TGR5 and TPH1 in the colon of the mice increased, leading to elevated 5‐HT levels. When the TGR5 gene was knocked out or the TPH1 inhibitor LX1606 was administered to suppress 5‐HT synthesis in constipated mice, the beneficial effects of Li05 on gastrointestinal motility and mucus secretion were reversed. Culturing intestinal organoids demonstrated that increased bile acids such as DCA, Iso‐LCA, and EALCA could enhance 5‐HT levels through the TGR5/TPH1 axis. Therefore, we concluded that Li05 regulated bile acid metabolism, subsequently increasing 5‐HT levels through the TGR5/TPH1 axis, thus alleviating constipation. The mechanism of Li05 in alleviating constipation. Li05 intervention regulated gut microbiota and bile acid metabolism, increasing 5‐HT levels through the TGR5/TPH1 activation to ameliorate constipation.

The effects of new strains of lactic acid bacteria on alfalfa silage fermentation were evaluated. The experiment was performed using a completely randomized design (with three replicates) based on a 6 × 6 factorial assay with 6 inoculants (I): Control (CTRL), Commercial inoculant (CI), Lactobacillus pentosus 14.7SE (LPE), Lactobacillus plantarum 3.7E (LP), Pediococcus pentosaceus 14.15SE (PP), and Lactobacillus plantarum 3.7E +  Pediococcus pentosaceus 14.15SE (LP + PP), and six fermentation periods (P): 1, 3, 7, 14, 28 and 56 days. Alfalfa was wilted for 6 h in the field, which increased the dry matter content to 368 g/kg as fed. The CP and yeast population decreased during the fermentation process. Silage inoculated with the PP strain had the lowest pH values beginning at 14 d of fermentation and the lowest acetic acid concentration on the last day of fermentation. New strains more efficiently regulated enterobacteria and mold populations at days 56 and 28, respectively. Silages inoculated with the PP strain had a higher coefficient of in vitro dry matter digestibility than LP silages. All of the tested novel strains resulted in positive effects on at least one chemical property of the silage during the fermentation process. However, the adding of P . pentosaceus can be indicated as the better for silage quality considering the tested treatments in the present study.

The present study describes the probiotic potential and functional properties of the lactic acid bacterium Pediococcus pentosaceus OBK05 isolated from buttermilk. The isolate OBK05 was assessed for its probiotic properties. The isolate showed notable tolerance to pH 2.0 and 3.0 (8.44, 8.35 log CFU/mL), oxbile of 0.5% at 2 and 4 h of incubation (6.97, 6.35 log CFU/mL) and higher aggregation (auto-aggregation, adhesion to hydrocarbons) than the referral strain, Lactobacillus acidophilus MTCC 10307. The adhesion efficiency to HT-29 cells was found to be maximum, corresponding to 93.5% and 97% at 1 and 2 h incubation, respectively. In addition, the isolate OBK05 showed antagonistic solid activity against bacterial pathogens like Pseudomonas aeruginosa MTCC 424 and Bacillus subtilis MTCC 1133. The phenotypic antibiotic resistance of the isolate was examined before and after curing plasmids. Among the known five structural genes responsible for different antibiotic resistance, four genes indicating antibiotic resistance to kanamycin- Aph (3´´)-III, streptomycin- strA , vancomycin- vanA and ciprofloxacin- gyrA were detected by PCR amplification of genomic DNA. Further, the horizontal gene transfer from OBK05 isolate to pathogens was not found for these antibiotic resistance markers when filter and food mating were carried out as no transconjugants developed on media plates containing respective antibiotics. This indicates that the intrinsic resistance is harbored on chromosomal genes, and hence it is nontransferable to other microbes. In addition, strain OBK05 exhibited good DPPH scavenging activity of 56 to 77% and liberated free amino acid from conjugated bile acid. The strain OBK05 demonstrated a strong ability to reduce cholesterol at 12 h (17%), 24 h (27%) and 48 h (67%) of incubation.

The biotransformation of L -glutamic acid ( L -Glu) to γ-aminobutyric acid (GABA) using glutamate decarboxylase (GAD) in microbial whole cells represents an ideal approach for biosynthesis of food-grade and pharmaceutical-grade GABA. To overcome the cell membrane barrier, enhance mass transfer efficiency in the whole-cell reaction system, and improve GABA biosynthesis efficiency, we established a novel ethanol-enhanced whole-cell biocatalytic co-reaction system through systematic investigations on the enzymatic characteristics of GAD in Pediococcus pentosaceus whole cells and the regulatory effects mediated by ethanol. The results showed that the optimal reaction pH and temperature for GAD in P. pentosaceus whole cells were 4.2 and 32 °C, respectively. A 7.5% (v/v) ethanol concentration significantly promoted the activity of whole-cell GAD, but reduced its stability. Through orthogonal test optimization, the optimal reaction conditions for ethanol-promoted GABA synthesis via P. pentosaceus whole-cell transformation were as follows: mixing 0.3 M monosodium glutamate (MSG) solution with 100 mg/ml cell suspension at a 1:1 volume ratio, adding 40 g/l of L -Glu/MSG (2:1) solid mixture, adjusting the final ethanol concentration to 3.75% (v/v), reacting at pH 4.2, 28 °C for 40 h. Under these conditions, the GABA yield reached 366.07 ± 5.57 mM, which was 21.44 ± 1.85% higher than that of the control group without ethanol. As an enhancer, ethanol demonstrates great application potential in GABA production via whole-cell transformation due to its high safety and ease of use.