Depiction of the In Vitro and Genomic Basis of Resistance to Hop and High Hydrostatic Pressure of Lactiplantibacillus plantarum Isolated from Spoiled Beer.

Among the beer-spoiling microorganisms, the dominant ones belong to the genera Lactobacillus, Leuconostoc, Oenococcus, and Pediococcus. It is assumed that resistance to hop bitters correlates with resistance to other factors and can significantly impact the brewing industry. Beer preservation with high hydrostatic pressure eliminates the spoiling microorganisms while preserving all desired properties of the beer. Here, we present comprehensive in vitro and genomic analysis of the beer-spoiling Lactiplantibacillus plantarum KKP 3573 capacity to resist hop and high hydrostatic pressure. Lp. plantarum KKP 3573 is a strain isolated from spoiled beer. Our finding suggests that the growth rate of the strain depends on the medium variant, where a small concentration of beer (5 IBU) stimulates the growth, suggesting that the limited concentration has a positive effect on cell growth. At the same time, increased concentrations of 20 IBU, 30 IBU, and pure beer 43.6 IBU decreased the growth rate of the KKP 3573 strain. We observed that higher extract content in the pressurized beer increased microbial survivability. The wort and Vienna Lager beer can stimulate the baroprotective effect. The taxonomy of the novel strain was confirmed after whole genome sequencing (WGS) and comparative genomic analysis. More specifically, it contains a chromosome of 3.3 Mb with a GC content of 44.4%, indicative of the Lp. plantarum species. Accordingly, it possesses high genomic similarity (>98%) with other species members. Annotation algorithms revealed that the strain carries several genes involved in resistance to stress, including extreme temperature, hop bitters and high pressure, and adaptation to the brewing environment. Lastly, the strain does not code for toxins and virulence proteins and cannot produce biogenic amines.

Complete Genome Sequencing and Bacteriocin Functional Characterization of Pediococcus ethanolidurans CP201 from Daqu.

This study aims to sequence the whole genome of Pediococcus ethanolidurans CP201 isolated from Daqu and determine the anti-corrosion ability of bacteriocins on chicken breast. The whole genome sequence information of P. ethanolidurans CP201 was analyzed, and its gene structure and function were explored. It was found that gene1164 had annotations in the NR, Pfam, and Swiss-Prot databases, and was related to bacteriocins. The exogenous expression of the bacteriocin gene Pediocin PE-201 was analyzed based on the pET-21b vector and the host BL21, and the corresponding bacteriocin was successfully expressed under the induction of IPTG. After purification by NI-NTA column, enterokinase treatment, membrane dialysis concentration treatment, and SDS-PAGE electrophoresis, the molecular weight was about 6.5 kDa and the purity was above 90%. By applying different concentrations of bacteriocin to chicken breast with different levels of contamination, the control of pathogenic bacteria, the ordinary contamination level (OC) group, and the high contamination level (MC) group could be completely achieved with 25 mg/L bacteriocin. In conclusion, the bacteriocin produced by the newly isolated CP201 can be applied to the preservation of meat products to prevent the risk of food-borne diseases.

NMR-Based Metabolomic Study on Phaseolus vulgaris Flour Fermented by Lactic Acid Bacteria and Yeasts.

In recent years, fermented foods have attracted increasing attention due to their important role in the human diet, since they supply beneficial health effects, providing important sources of nutrients. In this respect, a comprehensive characterization of the metabolite content in fermented foods is required to achieve a complete vision of physiological, microbiological, and functional traits. In the present preliminary study, the NMR-based metabolomic approach combined with chemometrics has been applied, for the first time, to investigate the metabolite content of Phaseolus vulgaris flour fermented by different lactic acid bacteria (LAB) and yeasts. A differentiation of microorganisms (LAB and yeasts), LAB metabolism (homo- and heterofermentative hexose fermentation), LAB genus (Lactobacillus, Leuconostoc, and Pediococcus), and novel genera (Lacticaseibacillus, Lactiplantibacillus, and Lentilactobacillus) was achieved. Moreover, our findings showed an increase of free amino acids and bioactive molecules, such as GABA, and a degradation of anti-nutritional compounds, such as raffinose and stachyose, confirming the beneficial effects of fermentation processes and the potential use of fermented flours in the production of healthy baking foods. Finally, among all microorganisms considered, the Lactiplantibacillus plantarum species was found to be the most effective in fermenting bean flour, as a larger amount of free amino acids were assessed in their analysis, denoting more intensive proteolytic activity.

Comparative genomics reveals key molecular targets for mutant Pediococcus pentosaceus C23221 producing pediocin.

Listeria monocytogenes is a common microorganism that causes food spoilage. Pediocins are some biologically active peptides or proteins encoded by ribosomes, which have a strong antimicrobial activity against L. monocytogenes. In this study, the antimicrobial activity of previously isolated P. pentosaceus C-2-1 was enhanced by ultraviolet (UV) mutagenesis. A positive mutant strain P. pentosaceus C23221 was obtained after 8 rounds of UV irradiation with increased antimicrobial activity of 1448 IU/mL, which was 8.47 folds higher than that of wild-type C-2-1. The genome of strain C23221 and wild-type C-2-1 was compared with identify the key genes for higher activity. The genome of the mutant strain C23221 consists of a chromosome of 1,742,268 bp, with 2052 protein-coding genes, 4 rRNA operons, and 47 tRNA genes, which is 79,769 bp less than the original strain. Compared with strain C-2-1, a total of 19 deduced proteins involved in 47 genes are unique to C23221 analyzed by GO database; the specific ped gene related to bacteriocin biosynthesis were detected using antiSMASH in mutant C23221, indicating mutant C23221 produced a new bacteriocin under mutagenesis conditions. This study provides genetic basis for further constituting a rational strategy to genetically engineer wild-type C-2-1 into an overproducer.

Genomic, metabolomic, and functional characterisation of beneficial properties of Pediococcus pentosaceus ST58, isolated from human oral cavity.

Bacteriocins produced by lactic acid bacteria are proteinaceous antibacterial metabolites that normally exhibit bactericidal or bacteriostatic activity against genetically closely related bacteria. In this work, the bacteriocinogenic potential of Pediococcus pentosaceus strain ST58, isolated from oral cavity of a healthy volunteer was evaluated. To better understand the biological role of this strain, its technological and safety traits were deeply investigated through a combined approach considering physiological, metabolomic and genomic properties. Three out of 14 colonies generating inhibition zones were confirmed to be bacteriocin producers and, according to repPCR and RAPD-PCR, differentiation assays, and 16S rRNA sequencing it was confirmed to be replicates of the same strain, identified as P. pentosaceus, named ST58. Based on multiple isolation of the same strain (P. pentosaceus ST58) over the 26 weeks in screening process for the potential bacteriocinogenic strains from the oral cavity of the same volunteer, strain ST58 can be considered a persistent component of oral cavity microbiota. Genomic analysis of P. pentosaceus ST58 revealed the presence of operons encoding for bacteriocins pediocin PA-1 and penocin A. The produced bacteriocin(s) inhibited the growth of Listeria monocytogenes, Enterococcus spp. and some Lactobacillus spp. used to determine the activity spectrum. The highest levels of production (6400 AU/ml) were recorded against L. monocytogenes strains after 24 h of incubation and the antimicrobial activity was inhibited after treatment of the cell-free supernatants with proteolytic enzymes. Noteworthy, P. pentosaceus ST58 also presented antifungal activity and key metabolites potentially involved in these properties were identified. Overall, this strain can be of great biotechnological interest towards the development of effective bio-preservation cultures as well as potential health promoting microbes.

Isolation, identification and safety evaluation of OTA-detoxification strain Pediococcus acidilactici NJB421 and its effects on OTA-induced toxicity in mice.

Ochratoxin A (OTA) is a potent mycotoxin found in foods and feeds, posing a health risk to animals and humans. Biological detoxification of OTA is considered a promising method, and some bacteria and fungi which can degrade OTA are isolated. However, research on safety and alleviating toxic effects are scarce. This study aims to isolate OTA-detoxification probiotics from natural samples and evaluate their safety and protective effects in mice. Here, a new OTA-detoxification strain named Pediococcus acidilactici NJB421 (P. acidilactici NJB421) was isolated from cow manure, which exhibited a removal rate of OTA at 48.53% for 48 h. P. acidilactici NJB421 exhibited high temperature resistance, acid tolerance, 0.3% bile salt and 1.4% trypsin resistance. The safety evaluation showed that P. acidilactici NJB421 at 2 × 108 CFU/per mouse had no abnormalities in body weight, organ indices, ALT, AST and ALP activities, BUN, CRE and TP contents. And P. acidilactici NJB421 alleviated the decreases in body weight, organ indices and small intestinal length, and alleviated intestinal injury, liver injury and kidney injury. These results suggest P. acidilactici NJB421 is safe and has protection against OTA poisoning, which provides a new OTA-detoxification strain for livestock and food industries.

Exploration of Antidiabetic, Cholesterol-Lowering, and Anticancer Upshot of Probiotic Bacterium Pediococcus pentosaceus OBK05 Strain of Buttermilk.

Chronic metabolic disorders such as hyperglycemia (diabetes), hyperlipidemia (high cholesterol), and cancer have become catastrophic diseases worldwide. Accordingly, probiotic intervention is a new approach for alleviating catastrophic diseases. In the present study, Pediococcus pentosaceus OBK05 was investigated as a potential probiotic bacterium for antidiabetic, cholesterol-lowering, and anticancer activities by in vitro and in vivo studies. Cell-free supernatant (CFS) of OBK05 showed potent antidiabetic activity by inhibition of α-amylase (72 ± 0.9%) and α-glucosidase (61 ± 0.8%) activity in vitro when compared to that of acarbose as standard (86 ± 0.9%). Furthermore, the in vivo cholesterol-lowering activity of OBK05 was studied using cholesterol-fed hypercholesterolemic mice. When compared to the cholesterol-fed group, the OBK05-intervened cholesterol-fed mice group had significantly lowered the levels of lipids and showed significantly lower lipid droplet accumulation in the histology of hepatocytes. Similarly, CFS exhibited higher anticancer activity (87.57 ± 1.27%) against HT-29 cells with an IC50 of 54.51 ± 1.8. Fluorescence-activated cell sorting (FACS) analysis revealed that CFS induced the cell cycle arrest by inhibiting the G1 to S phase transition. Further, in vivo anticancer activities were confirmed in BALB/c mice models, which were divided into four groups and held for 16 weeks. HT-29 cells (5×109/mice) were injected subcutaneously twice (0 and 2nd week), and OBK05 (3 ×105 CFU/mL) was given orally to the respective groups. The OBK05-intervened HT-29 cell-induced mice group showed significant improvement at the histological level and alleviated the degree of atypia in the intestine.

Genomic Analyses of Pediococcus pentosaceus ST65ACC, a Bacteriocinogenic Strain Isolated from Artisanal Raw-Milk Cheese.

Pediococcus pentosaceus ST65ACC was obtained from a Brazilian artisanal cheese (BAC) and characterized as bacteriocinogenic. This strain presented beneficial properties in previous studies, indicating its potential as a probiotic candidate. In this study, we aimed to carry out a genetic characterization based on whole-genome sequencing (WGS), including taxonomy, biotechnological properties, bacteriocin clusters and safety-related genes. WGS was performed using the Illumina MiSeq platform and the genome was annotated with the Prokaryotic Genome Annotation (Prokka). P. pentosaceus ST65ACC taxonomy was investigated and bacteriocin genes clusters were identified by BAGEL4, metabolic pathways were analyzed by Kyoto Encyclopedia of Genes and Genomes (KEGG) and safety-related genes were checked. P. pentosaceus ST65ACC had a total draft genome size of 1,933,194 bp with a GC content of 37.00%, and encoded 1950 protein coding sequences (CDSs), 6 rRNA, 55 tRNA, 1 tmRNA and no plasmids were detected. The analysis revealed absence of a CRISPR/Cas system, bacteriocin gene clusters for pediocin PA-1/AcH and penocin-A were identified. Genes related to beneficial properties, such as stress adaptation genes and adhesion genes, were identified. Furthermore, genes related to biogenic amines and virulence-related genes were not detected. Genes related to antibiotic resistance were identified, but not in prophage regions. Based on the obtained results, the beneficial potential of P. pentosaceus ST65ACC was confirmed, allowing its characterization as a potential probiotic candidate.

Enrichment and Distribution of Selenium in Pediococcus acidilactici MRS-7: Impact on Its Biochemical Composition, Microstructure, and Gastrointestinal Survival.

Lactic acid bacteria can convert selenium (Se) from inorganic to organic and elemental forms, but the distribution and existence form of organic Se in the bacteria are not clear after Se enrichment, and the effects of selenization on the growth and nutritional value of strains also need to be studied. In this study, Pediococcus acidilactici MRS-7 could absorb up to 67% of inorganic Se and convert most of it into organic Se; about 75% of organic Se was selenoprotein, 2.7% was Se-polysaccharide, and 4.6% was Se-nucleic acid. Additionally, Se-enriched treatment increased the levels of amino acids and essential elements in P. acidilactici MRS-7. Finally, after Se enrichment, Se nanoparticles (SeNPs) were found on the surface of P. acidilactici MRS-7, but they had no harmful effect on its morphology, and its survival during gastrointestinal digestion was not affected, indicating that SeP has potential probiotic value in the food industry.

Proteomic Profiling and Stress Response in Pediococcus acidilactici under Acetic Acid.

Lactic acid bacteria are indispensable functional microorganisms for cereal vinegar brewing, but cell activities are inhibited by the dominant acetic acid stress. Herein, an acetic-acid-tolerant strain isolated previously was identified as Pediococcus acidilactici, which also exhibited good resistance to other stresses during vinegar brewing. Proteomics analysis evidenced that differentially expressed proteins involved in the glycolysis and gluconeogenesis pathway, pyruvate metabolism, and sugar phosphotransferase system were all downregulated. Meanwhile, saturation of fatty acids and antioxidant enzymes was strengthened. The effects of several proteins on the resistance of P. acidilactici and Lactobacillus lactis relied on the types of strain and stress. AccA and AcpP participating in fatty acid metabolism and biosynthesis and Mnc related to stress response were found to protect cells by modifying fatty acid compositions and reinforcing the antioxidant defense system. Our works deepen the mechanisms of P. acidilactici under acetic acid and offer targets for engineering cell tolerance.

Evaluation of the Safety and Ochratoxin A Degradation Capacity of Pediococcus pentosaceus as a Dietary Probiotic with Molecular Docking Approach and Pharmacokinetic Toxicity Assessment.

The present study evaluated the properties and ochratoxin A (OTA) degradation capacity of the dietary probiotic Pediococcus pentosaceus BalaMMB-P3, isolated from a milk coagulant. The acidic tolerance of the isolate at pH 2-3 was checked with bile salts. No hemolytic activity was noted, which confirmed the nonpathogenicity of the strain. The isolate was tested in vitro for antibiotic susceptibility, enzymatic activity, bile salts hydrolase activity and antifungal activity against Penicillium verrucosum, Fusarium graminearum and Aspergillus ochraceus. A molecular docking-based OTA toxicity assessment was carried out for multitargeted proteins. The 16S rRNA gene-based phylogenetic assessment identified the strain as P. pentosaceus, and was authenticated in GenBank. The carboxylesterase and glutathione s-transferase enzymes showed active and strong interactions with esters and amide bonds, respectively. The compound exhibited carcinogenic and cytotoxicity effects at an LD50 value of 20 mg/kg. Furthermore, the strain showed a potent ability to reduce OTA and suggested the prospects for utilization in nutritional aspects of food.

Dynamics of the fermentation quality and microbiotsa in Ephedra sinica treated native grass silage.

AIMS: This study aimed to evaluate the effects of Ephedra sinica on physicochemical characteristics and bacterial community of ensiled native grass by multiple physicochemical analyses combined with high-throughput sequencing. METHODS AND RESULTS: Treatments were a control treatment with no additive (CON), E. sinica was added at 1% (CEa1), 3% (CEa2), and 5% of the fresh materials (CEa3). Compared to the CON group, the dry matter and water-soluble carbohydrate contents were significantly (p < 0.05) decreased in the CEa1 group. Compared to the CON group, the pH was significantly (p < 0.05) decreased in E. sinica treated silages, and a higher lactic acid content was observed in E. sinica treated silages. At the genus level, the abundance of Enterococcus, Lactobacillus, Pediococcus, and Weissella were the predominant member in the CON, CEa1, CEa2, and CEa3 groups, respectively. The abundance of Lactobacillus was significantly (p < 0.05) increased in the CEa1 group and Pediococcus was significantly (p < 0.05) increased in the CEa2 group. According to the 16S rRNA gene-predicted functional profiles, the inoculation of E. sinica accelerated the carbohydrate metabolism. CONCLUSIONS: In summary, the addition of E. sinica could improve the silage quality of native grass by regulating the bacterial community, and the addition of a 1% percentage of fresh materials exhibited the potential possibility of responding to get high-quality native grass silages. SIGNIFICANCE AND IMPACT OF THE STUDY: The utilization of herbal additives on fermentation quality combined with 16S rRNA gene-predicted functional analyses will contribute to the direction of future research in improving silage quality.

Evaluation of inhibitory compounds produced by bacteria isolated from a hydrogen-producing bioreactor during the self-fermentation of wheat straw.

AIMS: The objective of this study was to evaluate the inhibitory activity of compounds secreted by bacteria isolated from a hydrogen-producing bioreactor to understand how these microorganisms interact in this community. METHODS AND RESULTS: In vitro inhibitory assays were performed using samples secreted by bacteria subject to different treatments to determine if their inhibitory effect was due to organic acids, non-proteinaceous compounds or bacteriocin-like inhibitory substances (BLIS). Bacterial isolated were suppressed 43%, 30% and 27% by neutralized, precipitated and non-neutralized cell-free supernatants, respectively. Non-hydrogen producers (non-H2 P) lactic acid bacteria (LAB) (Lactobacillus plantarum LB1, Lactobacillus pentosus LB7, Pediococcus acidilactici LB4) and hydrogen producers (H2 P) LAB (Enterococcus faecium F) were inhibited by the production of organic acids, non-proteinaceous compounds and BLIS. Meanwhile, the obligate anaerobe H2 P (Clostridium beijerinckii B) inhibited by the production of non-proteinaceous compounds and BLIS. The presence of BLIS was confirmed when proteolytic enzymes affected the inhibitory activity of secreted proteins in values ranging from 20% to 42%. The BLIS produced by L. plantarum LB1, P. acidilactici LB4, L. pentosus LB7 and E. faecium F showed molecular masses of ~11, 25, 20 and 11 kDa, respectively. CONCLUSIONS: It was demonstrated antagonistic interactions between Lactobacillus-Enterococcus and Pediococcus-Enterococcus species, generated by the secretion of organic acids, non-proteinaceous compounds and BLIS. SIGNIFICANCE AND IMPACT OF THE STUDY: We report the interactions between LAB isolated from hydrogen-producing bioreactors. These interactions might impact the dynamics of the microbial population during hydrogen generation. Our work lays a foundation for strategies that allow controlling bacteria that can affect hydrogen production.

Heterologous expression of pediocin/papA in Bacillus subtilis.

Listeria monocytogenes is a food-borne pathogen. Pediocin is a group IIα bacteriocin with anti-listeria activity that is naturally produced by Pediococcus acidilactic and Lactobacillus plantarum. The pedA/papA gene encodes pediocin/plantaricin. In native hosts, the expression and secretion of active PedA/PapA protein rely on the accessory protein PedC/PapC and ABC transporter PedD/PapD on the same operon. The excretion machines were also necessary for pediocin protein expression in heterologous hosts of E. coli, Lactobacillus lactis, and Corynebacterium glutamicum. In this study, two vectors carrying the codon sequence of the mature PapA peptide were constructed, one with and one without a His tag. Both fragments were inserted into the plasmid pHT43 and transformed into Bacillus subtilis WB800N. The strains were induced with IPTG to secrete the fused proteins PA1 and PA2. Supernatants from both recombinant strains can inhibit Listeria monocytogenes ATCC54003 directly. The fused protein possesses inhibition activity as a whole dispense with removal of the leading peptide. This is the first report of active pediocin/PapA expression without the assistance of PedCD/PapCD in heterogeneous hosts. In addition, the PA1 protein can be purified by nickel-nitrilotriacetic acid (Ni-NTA) metal affinity chromatography.

Use of the ß-Glucan-Producing Lactic Acid Bacteria Strains Levilactobacillus brevis and Pediococcus claussenii for Sourdough Fermentation-Chemical Characterization and Chemopreventive Potential of In Situ-Enriched Wheat and Rye Sourdoughs and Breads.

The aim of the present study was to examine ß-glucan production and the potential prebiotic and chemopreventive effects of wheat and rye sourdoughs and breads generated with wild-type and non-ß-glucan-forming isogenic mutant strains of Levilactobacillus brevis and Pediococcus claussenii. Sourdough and bread samples were subjected to in vitro digestion and fermentation. Fermentation supernatants (FS) and pellets (FP) were analyzed (pH values, short-chain fatty acids (SCFA), ammonia, bacterial taxa) and the effects of FS on LT97 colon adenoma cell growth, viability, caspase-2 and -3 activity, genotoxic and antigenotoxic effects and on gene and protein expression of p21, cyclin D2, catalase and superoxide dismutase 2 (SOD2) were examined. Concentrations of SCFA were increased and concentrations of ammonia were partly reduced in the FS. The relative abundance of Bifidobacteriaceae was increased in all FPs. Treatment with FS reduced the growth and viability of LT97 cells and significantly increased caspase-2 and -3 activities without exhibiting genotoxic or antigenotoxic effects. The p21 mRNA and protein levels were increased while that of cyclin D2 was reduced. Catalase and SOD2 mRNA and protein expression were marginally induced. The presented results indicate a comparable chemopreventive potential of wheat and rye sourdoughs and breads without an additional effect of the formed ß-glucan.

Genomic and Phenotypic Evaluation of Potential Probiotic Pediococcus Strains with Hypocholesterolemic Effect Isolated from Traditional Fermented Food.

The use of probiotic microorganisms in food with the aim to confer health benefits to the host is one of the most critical roles of functional foods. Many pediococci bacteria frequently related to the meat environment, have technological properties, and are therefore commercially used as starter in the production of fermented meat products, such as different types of sausages. In this study, different lactic acid bacteria were isolated, identified to the species level, and then evaluated for their safety and functionality as possible probiotics. Different properties, such as resistance to simulated human gastrointestinal conditions, antimicrobial activity, and cholesterol-lowering effects, have been studied. Finally, the complete genome of one strain, namely P. acidilactici IRZ12B, which showed interesting features as a promising probiotic candidate, was sequenced and further studied. The results revealed that IRZ12B possesses interesting probiotic properties, particularly cholesterol-lowering capability and antimicrobial activity. In silico analysis evidenced the absence of plasmids, transmissible antibiotic resistance genes, and virulence factors. We also detected a bacteriocin encoding gene and a cholesterol assimilation-related protein. The phenotypical and genomic outcomes described in this study make P. acidilactici IRZ12B a very interesting cholesterol-lowering potential probiotic strain to be considered for the development of novel non-dairy-based functional foods.

Fermentation profile, cholesterol-reducing properties and chemopreventive potential of ß-glucans from Levilactobacillus brevis and Pediococcus claussenii - a comparative study with ß-glucans from different sources.

The aim of the present study was to investigate whether ß-glucans obtained from the lactic acid bacteria (LAB) Levilactobacillus (L.) brevis and Pediococcus (P.) claussenii exhibit similar physiological effects such as cholesterol-binding capacity (CBC) as the structurally different ß-glucans from oat, barley, and yeast as well as curdlan. After in vitro fermentation, fermentation supernatants (FSs) and/or -pellets (FPs) were analyzed regarding the concentrations of short-chain fatty acids (SCFAs), ammonia, bile acids, the relative abundance of bacterial taxa and chemopreventive effects (growth inhibition, apoptosis, genotoxicity) in LT97 colon adenoma cells. Compared to other glucans, the highest CBC was determined for oat ß-glucan (65.9 ± 8.8 mg g-1, p < 0.05). Concentrations of SCFA were increased in FSs of all ß-glucans (up to 2.7-fold). The lowest concentrations of ammonia (down to 0.8 ± 0.3 mmol L-1) and bile acids (2.5-5.2 µg mL-1) were detected in FSs of the ß-glucans from oat, barley, yeast, and curdlan. The various ß-glucans differentially modulated the relative abundance of bacteria families and reduced the Firmicutes/Bacteroidetes ratio. Treatment of LT97 cells with the FSs led to a significant dose-dependent growth reduction and increase in caspase-3 activity without exhibiting genotoxic effects. Though the different ß-glucans show different fermentation profiles as well as cholesterol- and bile acid-reducing properties, they exhibit comparable chemopreventive effects.

Structural analysis and biochemical properties of laccase enzymes from two Pediococcus species.

Prokaryotic laccases are emergent biocatalysts. However, they have not been broadly found and characterized in bacterial organisms, especially in lactic acid bacteria. Recently, a prokaryotic laccase from the lactic acid bacterium Pediococcus acidilactici 5930, which can degrade biogenic amines, was discovered. Thus, our study aimed to shed light on laccases from lactic acid bacteria focusing on two Pediococcus laccases, P. acidilactici 5930 and Pediococcus pentosaceus 4816, which have provided valuable information on their biochemical activities on redox mediators and biogenic amines. Both laccases are able to oxidize canonical substrates as ABTS, ferrocyanide and 2,6-DMP, and non-conventional substrates as biogenic amines. With ABTS as a substrate, they prefer an acidic environment and show sigmoidal kinetic activity, and are rather thermostable. Moreover, this study has provided the first structural view of two lactic acid bacteria laccases, revealing new structural features not seen before in other well-studied laccases, but which seem characteristic for this group of bacteria. We believe that understanding the role of laccases in lactic acid bacteria will have an impact on their biotechnological applications and provide a framework for the development of engineered lactic acid bacteria with enhanced properties.

Functions of small RNAs in Lactobacillus casei-Pediococcus group of lactic acid bacteria using fragment analysis.

Small RNAs (sRNA) are non-cording RNAs composed of 50∼400 nt responsible for coordinating the adaption of Escherichia coli and other bacteria to changing environmental conditions, including pH and temperature. However, the role of sRNAs in lactic acid bacteria (LAB) has not yet been clarified. In this study, we used the Lactobacillus casei-Pediococcus group to evaluate the function of sRNAs in LAB, using RNA sequencing in the exponential growth phase and stationary phase to map and analyze sRNA fragments, which were categorized as Pediococcus pentosaceus and Lactobacillus paracasei. We evaluated the role of sRNAs in nutrient synthesis for cell growth in exponential growth phase and in protein and biofilm biosynthesis for cell body durability. During exponential growth, the sRNA fragments were found to be involved in the stress response in Pediococcus pentosaceus and in environmental adaption in Lactobacillus paracasei. The results suggest that the function of sRNA can be characterized from sRNA fragments using RNA sequencing during the exponential growth and stationary phases in Lactobacillus casei-Pediococcus group.

Probiotic characterization of Pediococcus strains isolated from Iranian cereal-dairy fermented product: Interaction with pathogenic bacteria and the enteric cell line Caco-2.

In present study, we investigated the probiotic potential of three Pediococcus spp. isolated from Iranian traditional fermented cereal-dairy product, Tarkhineh. These 3 strains were identified as Pediococcus acidilactici VKU2, P. acidilactici IAH-5 and P. pentosaceus DHR005 by 16S rRNA gene sequencing. All the strain were found tolerate to pH 3 and 0.3% oxall for 3 h as well as simulated gastric and intestinal juice. P. acidilactici IAH-5 showed the highest cholesterol removal (67.52%), hydroxyl radical scavenging activity (58.32%), hydrophobicity (40.3%) and auto-aggregation (48%). Pediococcus spp. inhibited the growth of tested pathogens (Escherichia coli ATCC 25922, Pseudomonas aeruginosa PTCC 1707, Salmonella typhimurium PTCC 1609, and Staphylococcus aureus ATCC 25923) which the most susceptible strain was S. aureus. In competition assay, P. acidilactici IAH-5 was able to inhibited adhesion of 67.3% of S. typhimurium and in inhibition assay 45.8% of the pathogenic adhesion to Caco-2 cells were decreased. P. acidilactici VKU2 and P. acidilactici IAH-5 showed 16.32 and 12.25% adhesion to simulated epithelial cell line which were also confirmed by scanning electron microscopy. Pediococcus spp. did not showed DNase production or hemolytic activity which confirm its safety aspects. Our findings suggested that the P. acidilactici IAH-5 has the best properties with probiotic features and cholesterol assimilation for its application as novel bio-therapeutic and bio-preservation agents.