Search Results (100)

Acetate may act as a signaling molecule, regulating Paracin 1.7 production via quorum sensing (QS) in Lacticaseibacillus paracasei HD1.7. The “acetate switch” phenomenon requires mechanistic exploration to optimize Paracin 1.7 production. The “acetate switch” phenomenon delays with higher glucose levels (30 h, 36 h, and 96 h). Before the occurrence of the “acetate switch”, the ATP content increases and peaks at the “acetate switch” point and the NAD⁺/NADH ratio decreases, indicating energy changes. Moreover, the QS genes used for the pre-regulation of bacteriocin, such as prcKR, comCDE, were highly expressed. After the “acetate switch”, the ATP content decreased and the QS genes for the post-regulation of bacteriocin were highly expressed, such as rggs234 and sigma70-1/70-2. The “acetate switch” could act as an energy switch, regulating bacterial growth and QS genes. Before and after the “acetate switch”, some metabolic pathways were significantly altered according to the transcriptomic analysis by HD1.7 and HD1.7-Δpta. In this study, acetate was used as an input signal to regulate the two-component system, significantly influencing the bacteriocin expression system. And this study clarifies the roles of acetate, energy, and quorum sensing in promoting Paracin 1.7 production, providing a theoretical basis for optimizing the bacteriocin fermentation process of HD1.7. Full article

The emergence of multidrug-resistant pathogens has driven the development of novel antimicrobial peptides (AMPs) as therapeutic alternatives. Lactolisterin LBU (LBU) is a bacteriocin with promising activity against Gram-positive bacteria, including Staphylococcus aureus. In this study, we designed and evaluated a panel of amino acid variants of LBU to investigate domain–activity relationships and improve activity. Peptides were commercially synthesized, and their effect was evaluated for minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC), hemolytic activity, cytotoxicity, in vivo toxicity, and virulence modulation. AlphaFold3 structural prediction of LBU revealed a four-helix topology with amphipathic and hydrophobic segments. Helical wheel projections identified helices I and IV as amphipathic, suggesting their potential involvement in membrane interaction and activity. Glycine-to-alanine substitutions at helix I markedly increased antimicrobial activity but altered toxicity profiles. In contrast, changes at helix junctions and kinks reduced antimicrobial activity. We also showed differential regulation of virulence genes upon sub-MIC treatment. Overall, rational substitution enabled identification of residues critical for activity and toxicity, providing insights into therapeutic tuning of lactolisterin-based peptides. Full article

Antimicrobial food packaging is considered a promising technology to improve food safety by inhibiting or reducing the growth of food microorganisms and minimizing the need for preservatives. This study aimed to develop and evaluate carboxymethyl cellulose (CMC) films integrated with bacteriocins for antibacterial efficacy. Plantaricin W was assessed as a potential bacteriocin for activation of CMC to control the dangerous food-borne pathogen, Listeria monocytogenes. Minced beef samples were inoculated with L. monocytogenes ATCC BAA-679 and treated with plantaricin W-activated food packaging. The results showed a significant reduction of the target pathogen by approximately 1 log cycle compared to the control group. Enterocin F4-9 is a novel bacteriocin that acts on Gram-negative microbes that were not affected by plantaricin W. Therefore, a novel food packaging activated with plantaricin W and enterocin F4-9 was developed to broaden their antimicrobial activity. The effect of this film on meat-associated microbes was investigated. The results demonstrated that the film significantly reduced the counts of mesophilic and psychotropic bacteria by 86.67% and 96.67%, respectively. Additionally, the pH values of the treated meat samples were significantly lower than those of the untreated controls. The obtained findings indicated that bacteriocin-activated CMC films could potentially be utilized as antimicrobial packaging in modern food technology. Full article

Soil, rhizosphere, and plant-associated microorganisms can enhance plant growth and health. A genomic analysis of these microbes revealed the key characteristics contributing to their beneficial effects. Following a field survey in Panama, four bacterial isolates with plant growth-promoting traits (PGPT) in rice (Oryza sativa L.) were identified. In this study, we sequenced, assembled, and annotated the genomes of Lysinibacillus fusiformis C6 and 24, and Bacillus cereus D23 and 59. The C6 genome was 4,754,472 bp long with 10 contigs, 37.62% guanine-cytosine (GC) content, and 4657 coding sequences (CDS). The 24 genome was 4,683,219 bp with five contigs, 37.65% GC content, and 4550 CDS. The D23 genome was 6,199,908 bp long with 18 contigs, 34.84% GC content, and 6141 CDS. The 59 genome was 6,194,462 bp with 21 contigs, 34.87% GC content, and 6122 CDS. Digital DNA–DNA hybridization (dDDH) and average nucleotide identity (ANI) confirmed that C6 and 24 belong to Lysinibacillus fusiformis, whereas D23 and 59 belong to the Bacillus cereus species. Further results revealed that these bacteria contained genes characteristic of plant growth-promoting bacteria, such as siderophore, phytohormone auxin (IAA) production, and nitrogen-fixing abilities that promote plant growth. Moreover, the antiSMASH database identified gene clusters involved in secondary metabolite production (biosynthetic gene clusters), such as betalactone, NRPS-like, NRP-siderophore, terpene, and RiPP-like clusters. Moreover, diverse and novel biosynthetic clusters (BCGs) have included non-ribosomal peptides (NRPs), polyketides (PKs), bacteriocins, and ribosomally synthesized and post-transcriptionally modified peptides (RiPPs). This work offers new insights into the genomic basis of the studied strains’ plant growth-promoting capabilities. Full article

We hypothesize that bacteria isolated from free-ranging animals could potentially be useful for practical applications. To meet this objective a Gram-positive bacterium was isolated from the gastrointestinal (GI) tract of a Gray Wolf (Canis lupus) using Brucella broth with hemin and vitamin K (BBHK). By small ribosomal RNA (16S) gene sequencing the bacterium was initially identified as a novel Carnobacterium maltaromaticum strain. The bacterium could be propagated both anaerobically and aerobically and was both catalase/oxidase negative and negative by the starch hydrolysis as well as negative using lipase assays. The reference whole genome sequence (WGS) was obtained using both Illumina and Nanopore sequencing. The genome assembly was 3,512,202 bp in length, encoding core bacterial genes with a GC% content of 34.48. No lysogenic bacteriophage genes were detected, although the genome harbors genes for the expression of bacteriocin and other secondary metabolites with potential antimicrobial properties. Multilocus sequence typing (MLST), WGS phylogenetics, average nucleotide identity (ANI), and single nucleotide polymorphism (SNP) analyses of the isolate’s genome indicate this bacterium is a newly identified Carnobacterium maltaromaticum sequence type (ST). Members of the Carnobacteria have anti-listeria activities, highlighting their potential functional properties. Consequently, the isolate could be a potential probiotic for canids and this is the first report on an axenic C. maltaromaticum culture from the genus Canis. Full article

Microorganisms exert antagonistic effects on pathogens through different mechanisms, thereby achieving biological control of plant diseases. Many Burkholderia strains can produce complex secondary metabolites and substances that have toxic effects on host cells. The phage tail-like bacteriocins (tailocins) is a compound with antibacterial activity. However, its function in B. multivorans has not yet been reported. This article explores the ability of B. multivorans WS-FJ9 to antagonise plant pathogenic fungi and oomycetes, screening the potential tailocins in the strain WS-FJ9 and verifying their function, to reveal its novel antimicrobial mechanisms. We found that WS-FJ9 had strong antagonistic effects on the plant pathogenic fungi Phomopsis macrospore and Sphaeropsis sapinea, and the pathogenic oomycete Phytophthora cinnamomi. The phage tail-like protein Bm_67459 was predicted from the WS-FJ9 strain genome. The Bm_67459 cDNA encoded 111 amino acid sequence, and the relative molecular weight was approximately 11.69 kDa, the theoretical isoelectric point (pI) was 5.49, and it was a hydrophilic protein. Bm_67459 had no transmembrane helix region or signal peptide, and it belonged to the Phage_TAC_7 super family. qRT-PCR results showed that Bm_67459 gene expression was significantly upregulated during contact between WS-FJ9 and P. cinnamomi. The purified Bm_67459 protein significantly inhibited P. cinnamomi mycelial growth at 10 μg·mL⁻¹. In summary, the WS-FJ9 strain had broad-spectrum anti-phytopathogenic activity, and the tailocin Bm_67459 was an important effector against the plant pathogen P. cinnamomi, which helps to reveal the antagonistic mechanism of this strain at the molecular level and provides excellent strain resources for the biological control of plant diseases. Full article

Bacteriocins, ribosomally synthesized by bacteria, have long been recognized for their role in ensuring food safety and security due to their antibacterial effects against foodborne pathogens and spoilage bacteria. However, recent advancements have unveiled their expanding potential beyond food applications, with increasing evidence of their efficacy against clinically significant pathogenic bacteria, biofilm formation, viral infections, and even cancer. These emerging discoveries have continuously added new layers to the application of bacteriocins, extending their relevance from food preservation to broader human health interventions. To further harness this expanding potential, various innovative strategies have been developed to overcome traditional limitations associated with bacteriocin use. Instead of directly employing bacteriocins or bacteriocin-producing bacterial cultures, novel approaches, such as incorporating them into films and packaging materials or coupling them with nanoparticles, have demonstrated enhanced effectiveness. In this review, we examine the evolving landscape of bacteriocin applications and shed light on the expanding functional spectrum of bacteriocins for both food safety and human health, although some important challenges and limitations remain. By analyzing the recent literature and innovative technological advancements, we highlight how bacteriocins are continuously evolving, opening new frontiers for their use and reinforcing their significance beyond their conventional roles. Full article

Circular bacteriocins are potent antimicrobials against pathogenic Gram-positives. In searching for marine bacteriocins, an antibacterial peptide (flexusin A) was purified from the fermentation broth of marine bacterium Bacillus flexus R29-2. Genome sequencing and gene annotation revealed the chromosome contained an unknown circular bacteriocin gene cluster. Approaches including shot-gun proteomics analysis, AntiSMASH and BAGEL4 predication as well as the comprehensive sequence alignment, were then conducted, respectively, to verify the correlation of flexusin A with the gene-encoded precursor peptide. The results confirmed that flexusin A was the mature circular bacteriocin of the predicated precursor peptide with six amino acids as leader peptide. Flexusin A was 6098.4 Da in size, with a net charge of +3 and PI of 9.60. It shared the typical saposin-like fold spatial conformation features as commonly found in other circular bacteriocins. Flexusin A was pH, thermal, and protease tolerant. It exhibited a narrow antimicrobial spectrum against Gram-positives, and it can strongly inhibit Staphylococcus aureus by causing cell destruction via membrane destabilization. Taken together, a novel circular bacteriocin flexusin A was identified in this work. The characterization of flexusin A has extended circular bacteriocins family to 26 members. This is also the first report on bacteriocin production by B. flexus. Full article

Proteus mirabilis has been identified as the third most frequent reason for catheter-associated urinary tract infections. The production of urease significantly enhances the force of catheter blockage caused by biofilm formation. Because biofilms are important virulence factors that make antibiotics less potent, it is becoming increasingly important to develop novel alternative antibiotics. In addition to the unique properties they possess, nanoparticles made from metal oxide are currently attracting considerable attention as possible antibacterials. This research aims to explore the potential anti-biofilm properties of green manufactured ZnO-CuO nanoparticles generated by P. mirabilis. By synthesizing reductive enzymes, bacterial cells can participate in the biosynthesis process. This study explores whether green synthesized ZnO-CuO nanoparticles can work as an anti-biofilm agent formed by P. mirabilis. These nanoparticles were generated using Bacteriocins to determine their effectiveness against bacteria, which were partially purified and showed antimicrobial activity against Gram-negative bacteria of P. mirabilis. AFM, TEM, FESEM, XRD, and ultraviolet (UV)–visible spectroscopy were used to analyze the biosynthesized nanoparticles to ascertain their chemical and physical characteristics. XRD verified the hexagonal structure, TEM demonstrated a size range of 96.00 nm, and FESEM verified the surface morphology. The dispersion and roughness of the nanoparticles are shown through AFM examination. The produced nanoparticles’ UV-visible spectra displayed a maximum peak at 287 and 232 nm. When applied to strains (wild-type) of Proteus mirabilis (multidrug-resistant), copper and zinc nanoparticles had notable biofilm inhibitory effects. Weak biofilm production has been demonstrated by bacteria that effectively generate biofilms, following incubation with 128 μg/mL subminimum inhibitory concentrations (MICs) of CuO nanoparticles for 24 and 48 h at 37 °C. Following treatment with the ZnO-CuO nanocomposite of these strains, downregulation alterations in LuxS expression were detected by utilizing a real-time PCR process. After this, isolates were treated with the nanocomposite, and downregulated shifts in LuxS expression were found by utilizing the real-time PCR technique in contrast with the isolates that were not treated. Zinc oxide (ZnO) nanoparticles can be utilized as antibacterial agents in a concentration-dependent manner, aligning with all observed findings. The present research demonstrates that green synthesized copper oxide–zinc oxide nanocomposites are effective anti-biofilm agents against P. mirabilis. Their noteworthy downregulation of LuxS gene expression successfully prevents biofilm formation and swarming motility. Full article

Background/objectives: Bacteriocins can be considered a novel source of natural alternatives to antibiotics or chemical food additives with the potential to fight against clinical and food pathogens. A number have already been commercialised as food preservatives, but they also have the potential to treat drug-resistant clinical pathogens and can play a role in immune modulation. To achieve their full potential, an understanding of their mode of action is required. Methods: Bactofencin A and nisin A were purified to homogeneity by reversed-phase HPLC and their effect on the mastitis pathogen Staphylococcus aureus DPC5246 was assessed by cell viability assays and flow cytometry. Results: We report that bactofencin A displays a delayed inhibitory effect against the mastitis pathogen, Staphylococcus aureus DPC5246, suggesting an unusual mode of action. This characteristic was clearly visible on BHI plate media, where formation of inhibition zones against the staphylococcal strain took 23 h compared to 6 h for the well-characterised nisin. This delayed killing and injury was also demonstrated using flow cytometry, where damage was evident 4 h after bacteriocin addition. Treatment with 2 μM bactofencin A resulted in approximately 20-fold higher numbers of injured and 50-fold higher numbers of dead cells when compared to untreated cells. Combining bactofencin A with the lantibiotic nisin A resulted in faster killing at lower bacteriocin concentrations. When combined in an equal ratio, the combination exhibited a 4-fold increase in inhibition compared to nisin A alone. These results demonstrate that the combination may be very effective in therapeutic applications against pathogenic staphylococci. Full article

Listeria monocytogenes is a Gram-positive bacterium causing listeriosis, a severe infection responsible for significant morbidity and mortality globally. Its persistence on food processing surfaces via biofilm formation presents a major challenge, as conventional sanitizers and antimicrobials exhibit limited efficacy against biofilm-embedded cells. This study investigates a novel approach combining an engineered polysaccharide-degrading enzyme (CAase) with a bacteriocin (thermophilin 110) produced by Streptococcus thermophilus. Laboratory assays evaluated the effectiveness of this combination in disrupting biofilms and inactivating L. monocytogenes on various surfaces. The results demonstrated that CAase effectively disrupts biofilm structures, while thermophilin 110 significantly reduces bacterial growth and viability. The preliminary trials indicate a dual-action approach offers a potential alternative to conventional treatments, enhancing food safety by effectively controlling Listeria biofilms in food processing environments. Full article

Bacteriocins are antimicrobial peptides produced by bacteria with promising potential for controlling pathogens in various fields. This study highlights recent advances in the research on bacteriocins, providing a comprehensive overview of emerging technologies applied to the production and stability of these compounds, as the use of alternative substrates and encapsulation techniques. In recent decades, significant efforts have focused on discovering novel molecules with broad-spectrum activity capable of combating both Gram-positive and Gram-negative microorganisms, including clinically and industrially relevant pathogens. Recent studies explore strategies to optimize bacteriocin production, such as modifications in cultivation parameters aimed at reducing costs and increasing yield. Additionally, microencapsulation techniques have been widely discussed, emphasizing their role in enhancing the stability and efficacy of bacteriocins under adverse conditions. Finally, this article examines the potential applications of bacteriocins, highlighting their use as natural food preservatives, therapeutic alternatives for infection control, and bioactive agents in sustainable agriculture. These advancements establish bacteriocins as versatile agents with significant technological and economic impacts. Full article

Background/Objectives: Antimicrobial-resistant (AMR) pathogens represent a serious threat to public health, particularly in food production systems where antibiotic use remains widespread. As a result, alternative antimicrobial treatments to antibiotics are essential for effectively managing bacterial infections. This study aimed to identify and characterize novel antimicrobial peptides produced by bacteria, known as bacteriocins, as well as to recognize safe bacteriocin-producing strains, sourced from poultry slaughterhouse effluents. Methods: A total of 864 bacterial isolates were collected across eight stages of a poultry slaughter line and screened for antimicrobial activity against Gram-positive and Gram-negative indicator strains. Whole-genome sequencing (WGS) was performed on 12 selected strains, including Enterococcus faecium (6 isolates), Lactococcus lactis (1 isolate), Lactococcus garvieae (1 isolate) and Escherichia coli (4 isolates). The presence of bacteriocin gene clusters (BGC), antibiotic resistance genes (ARG), and virulence factors (VF) was analyzed. The antimicrobial activity of a novel bacteriocin was further evaluated using in vitro cell-free protein synthesis (IV-CFPS). Results: WGS revealed multiple BGCs, including a novel class IId bacteriocin, lactococcin P1A (LcnP1A), in L. lactis SWD9. LcnP1A showed antimicrobial activity against various indicator strains, including Listeria monocytogenes. While most bacteriocin-encoding strains harbored ARGs and VFs, E. faecium SWG6 was notable for its absence of ARGs and minimal VFs, highlighting its potential as a probiotic. Conclusions: These findings underscore the importance of discovering novel bacteriocins and safer bacteriocin producing strains to address antimicrobial resistance in the food chain. Further research would validate the efficacy of both the novel lactococcin P1A bacteriocin and the E. faecium SWG6 isolate for application in processed food and animal production systems. Full article

Antimicrobial resistance (AMR) poses a significant challenge to animal production due to the widespread use of antibiotics. Therefore, there is an urgent need for alternative antimicrobial strategies to effectively manage bacterial infections, protect animal health, and reduce reliance on antibiotics. This study evaluated the use of emerging approaches and procedures for the isolation, identification, and characterization of bacteriocin-producing bacteria and their bacteriocins, sourced from the gastrointestinal tract (GIT) of meat-producing pigs. Out of 2056 isolates screened against Gram-positive and Gram-negative indicator strains, 20 of the most active antimicrobial isolates were subjected to whole genome sequencing (WGS) for the prediction of coding DNA sequences (CDS) and the identification of bacteriocin gene clusters (BGC) and their functions. The use of an in vitro cell-free protein synthesis (IV-CFPS) protocol and the design of an IV-CFPS coupled to a split-intein mediated ligation (IV-CFPS/SIML) procedure made possible the evaluation of the production and antimicrobial activity of described and putatively novel bacteriocins. A colony MALDI-TOF MS procedure assisted in the identification of class I, II, and III lanthipeptides. MALDI-TOF MS and a targeted proteomics, combined with a massive peptide analysis (LC-MS/MS) approach, has proven valuable for the identification and biochemical characterization of previously described and novel bacteriocins encoded by the isolated bacteriocin-producing strains. Full article

Shouchella clausii (S. clausii) has been marketed as an important commercial probiotic, displaying significant therapeutic effects on antibiotic-associated diarrhea and providing benefits to humans. This study aimed to explore the distribution, adaptation, and probiotic properties of S. clausii. Based on 16S rRNA gene analysis, 43 strains of S. clausii were isolated from 317 soil samples in China. Based on the genomic index of Average Nucleotide Identity (ANI) results, 41 strains were confirmed as S. clausii, while two strains, FJAT-45399 and FJAT-45335, were identified as potential novel species distinct from S. clausii. Combined phenotypic and genomic predictions indicated that S. clausii could survive under harsh conditions. Comparative genomics revealed that these isolates possess antibiotic resistance genes, as well as capabilities for bacteriocin and folate production, while lacking toxins and hemolytic activity. Hemolysis tests indicated that strain FJAT-41761 exhibited non-pathogenic γ-hemolytic activity, while also demonstrating resistance to multiple antibiotics, consistent with probiotic characteristics. These findings suggest that strain FJAT-41761 is safe and holds potential as a future probiotic. Full article