Metabolic and protein expression responses of Shewanella baltica in golden pomfret broths to slightly acidic electrolysed water.

Shewanella baltica is a specific spoilage organism of golden pomfret. This study aims to explore the antibacterial mechanism of slightly acidic electrolysed water (SAEW) against S. baltica (strains ABa4, ABe2 and BBe1) in golden pomfret broths by metabolomics, proteomics and bioinformatics analyses. S. baltica was decreased by at least 3.94 log CFU/mL after SAEW treatment, and strain ABa4 had the highest resistance. Under SAEW stress, amino acids and organic acids in S. baltica decreased, and nucleotide related compounds degraded. Furthermore, 100 differentially expressed proteins (DEPs) were identified. Most DEPs of strains ABe2 and BBe1 were down-regulated, while some DEPs of strain ABa4 were up-regulated, especially those oxidative stress related proteins. These results suggest that the modes of SAEW against S. baltica can be traced to the inhibition of amino acid, carbon, nucleotide and sulphur metabolisms, and the loss of functional proteins for temperature regulation, translation, motility and protein folding.

Progress in antibacterial applications of nanozymes.

Bacterial infections are a growing problem, and antibiotic drugs can be widely used to fight bacterial infections. However, the overuse of antibiotics and the evolution of bacteria have led to the emergence of drug-resistant bacteria, severely reducing the effectiveness of treatment. Therefore, it is very important to develop new effective antibacterial strategies to fight multi-drug resistant bacteria. Nanozyme is a kind of enzyme-like catalytic nanomaterials with unique physical and chemical properties, high stability, structural diversity, adjustable catalytic activity, low cost, easy storage and so on. In addition, nanozymes also have excellent broad-spectrum antibacterial properties and good biocompatibility, showing broad application prospects in the field of antibacterial. In this paper, we reviewed the research progress of antibacterial application of nanozymes. At first, the antibacterial mechanism of nanozymes was summarized, and then the application of nanozymes in antibacterial was introduced. Finally, the challenges of the application of antibacterial nanozymes were discussed, and the development prospect of antibacterial nanozymes was clarified.

Construction of BSA-ZnO&Quercetin based multifunctional bionic self-assembly system and their antibacterial mechanism study.

The misuse of antibiotics has led to the growing problem of multidrug-resistant (MDR) bacteria, and there is still a lack of effective antibacterial agents that can replace antibiotics. Therefore, the design and development of multifunctional nanomaterials with long-term inhibitory effects on drug-resistant bacteria are extremely challenging. In this study, a multifunctional biomimetic self-assembly system, BSA-ZnO&Quercetin, based on bovine serum albumin (BSA), ZnO, and quercetin, was established using a simple and controllable method. The prepared self-assembly system has high stability and biocompatibility, and could fully combine the performance advantages of each component. BSA-ZnO&Quercetin showed excellent broad-spectrum antibacterial activity without inducing bacterial resistance. The related antibacterial mechanism of BSA-ZnO&Quercetin primarily involves biofilm inhibition and destruction, and inducing the production of reactive oxygen species, resulting in the death of the bacteria. The biomimetic self-assembly system BSA-ZnO&Quercetin constructed in this research is expected to replace antibiotics for antibacterial application.

Antibacterial Activity and Mechanism of Taxillµs chinensis (DC.) Danser and Its Active Ingredients.

Taxillµs chinensis (DC.) Danser is a traditional Chinese herbal medicine. It has not been reported regarding antibacterial active ingredients and mechanisms of action. However, the Chinese patent medicine Yinhua Miyanling Tablets containing Taxillµs chinensis has an obvious anti-infective effect in our patent. Therefore, we speculate that Taxillµs chinensis may have antibacterial activity. The purpose of this paper is to study the antibacterial effect and mechanism of Taxillµs chinensis and find active compounds with antibacterial activity and a mechanism. We studied the antibacterial effect and mechanism of Taxillµs chinensis extract. The compounds in the ethyl acetate extract of Taxillµs chinensis were preliminarily identified by UPLC-Q-Orbitrap and analyzed by mass spectrometry. Above all, the antibacterial effect and antibacterial mechanism of the active components of Taxillµs chinensis were determined. Finally, we found, for the first time, that Taxillµs chinensis has a good antibacterial effect and ethyl acetate extract has the best effect. In addition, we found, for the first time, that it has an active component, 4-indolecarbaldehyde, and the component has a good broad-spectrum antibacterial effect. Above all, the active chemical 4-indolecarbaldehyde of Taxillµs chinensis can destroy the bacterial structure, make it unable to maintain normal morphology, and significantly increase the number of deaths. In short, Taxillµs chinensis has an antibacterial effect, and one of its main antibacterial components is 4-indolecarbaldehyde. The antibacterial mechanism of Taxillµs chinensis and 4-indolecarbaldehyde is related to the change in bacterial membrane permeability.

Antibacterial mechanism of atmospheric cold plasma against Pseudomonas fluorescens and Pseudomonas putida and its preservation application on in-packaged red shrimp paste.

This study aimed to investigate the antibacterial mechanism of atmospheric cold plasma (ACP) against Pseudomonas fluorescens and Pseudomonas putida and its preservation effect on red shrimp paste. A reactive species (RS) assay showed that O3, H2O2, and total nitric oxide were generated after ACP treatment, which possessed great potential for antibacterial and food preservation. In vitro antibacterial results showed that excess RS inhibited bacterial activity through cell membrane damage. Molecular docking predictions and enzyme activity assays indicated that ACP-induced RS might deactivate dehydrogenases (such as malic dehydrogenase) by oxidatively modifying the active sites. Fluorescence quantification experiments validated the damage of RS to dsDNA. Further preservation tests on shrimp paste demonstrated that ACP treatment significantly delayed the increase in total viable count, Pseudomonas count, and total volatile basic­nitrogen during refrigeration. This study deepened the understanding of the antibacterial mechanism of ACP and highlighted its potential application as a new preservation method.

Antimicrobial Peptides Derived from Bacteria: Classification, Sources, and Mechanism of Action against Multidrug-Resistant Bacteria.

Antimicrobial peptides (AMPs) are short, usually cationic peptides with an amphiphilic structure, which allows them to easily bind and interact with the cellular membranes of viruses, bacteria, fungi, and other pathogens. Bacterial AMPs, or bacteriocins, can be produced from Gram-negative and Gram-positive bacteria via ribosomal synthesis to eliminate competing organisms. Bacterial AMPs are vital in addressing the increasing antibiotic resistance of various pathogens, potentially serving as an alternative to ineffective antibiotics. Bacteriocins have a narrow spectrum of action, making them highly specific antibacterial compounds that target particular bacterial pathogens. This review covers the two main groups of bacteriocins produced by Gram-negative and Gram-positive bacteria, their modes of action, classification, sources of positive effects they can play on the human body, and their limitations and future perspectives as an alternative to antibiotics.

TMT-Based Quantitative Proteomics Revealed the Antibacterial Mechanism of Cinnamaldehyde against MRSA.

Natural plant extracts have demonstrated significant potential in alternative antibiotic therapies. Cinnamaldehyde (CA) has garnered considerable attention as a natural antibacterial agent. In this study, Tandem mass tag (TMT) quantitative proteomics combined with Western blot and RT-qPCR methods were employed to explore the antibacterial mechanism of CA against Methicillin-Resistant Staphylococcus aureus (MRSA) at the protein level. The results showed that a total of 254 differentially expressed proteins (DEPs) were identified in the control group and CA treatment group, of which 161 were significantly upregulated and 93 were significantly downregulated. DEPs related to nucleotide synthesis, homeostasis of the internal environment, and protein biosynthesis were significantly upregulated, while DEPs involved in the cell wall, cell membrane, and virulence factors were significantly downregulated. The results of GO and KEGG enrichment analyses demonstrated that CA could exert its antibacterial effects by influencing pyruvate metabolism, the tricarboxylic acid (TCA) cycle, teichoic acid biosynthesis, and the Staphylococcus aureus (S. aureus) infection pathway in MRSA. CA significantly inhibited the expression of recombinant protein MgrA (p < 0.05), significantly reduced the mRNA transcription levels of mgrA, hla, and sdrD genes (p < 0.05), and thermostability migration assays demonstrated that CA can directly interact with MgrA protein, thereby inhibiting its activity. These findings suggest that CA exerts its antibacterial mechanism by regulating the expression of related proteins, providing a theoretical basis for further development of clinical applications of antimicrobial agents derived from natural plant essential oils in the treatment of dairy cow mastitis.

Antibacterial Activity and Cytotoxicity of the Novel Bacteriocin Pkmh.

Bacteriocins are a class of proteins produced by bacteria that are toxic to other bacteria. These bacteriocins play a role in bacterial competition by helping to inhibit potential competitors. In this study, we isolated and purified a novel bacteriocin Pkmh, different from the previously reported bacteriocin PA166, from Pseudomonas sp. strain 166 by ammonium sulfate precipitation, dialysis membrane method, ion exchange chromatography, and gel filtration chromatography. SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) revealed that the molecular weight of Pkmh is approximately 35 kDa. Pkmh exhibited potent antimicrobial activity against bovine Mannheimia haemolytica (M. haemolytica) with low cytotoxicity, and lower hemolytic activity was observed. In addition, Pkmh retained antimicrobial activity at different pH ranges (2-10) and temperature conditions (40, 60, 80, 100 °C). Our analysis of its antimicrobial mechanism showed that Pkmh acts on bacterial cell membranes, increasing their permeability and leading to cell membrane rupture and death. In conclusion, Pkmh exhibited low hemolytic activity, low toxicity, and potent antibacterial effects, suggesting its potential as a promising candidate for clinical therapeutic drugs.

Antibacterial evaluation of garlic extracts on chitosan/starch packaging film using response surface methodology and its application for shelf-life extension of bell peppers (Capsicum annuum).

In this study, garlic extract (GE) was assessed as a potential additive in chitosan/starch (Ch/De) coatings, focusing on phenolic and flavonoid content analyses and antibacterial properties. Using response surface methodology approach, an optimization method was employed to achieve the optimal antibacterial formulation, with Ch, De, and GE identified as key variables in the Design of Experiment. Fourier transform infrared spectroscopy and X-ray diffraction analyses elucidated interactions among these primary components within the films, while thermogravimetric analysis confirmed the enhanced thermal stability of GE-coated film formulations (Ch/De/GE). The Ch/De/GE exhibited antibacterial efficacy against Escherichia coli (ATCC 25922) with an inhibition zone of 7.2 mm at optimized concentrations of 2% w/v Ch, 1.5% w/v starch, and 0.5% v/v GE. In silico molecular docking studies provided insights into GE's inhibitory role as an antibacterial agent. Evaluation of green and yellow bell peppers (Capsicum annuum) over 18 days showed that coated peppers maintained better visual appearance and mass stability, with a weight loss decrease of 40.54%-48.96%, compared to uncoated ones. Additionally, the Ch/De/GE coating effectively inhibited bacterial growth, reducing it by 1-1.23 log CFU, during the storage period. In conclusion, the Ch/De/GE coating effectively extends the shelf-life of bell peppers and maintains their quality, demonstrating its potential for use in food packaging to preserve perishable items. PRACTICAL APPLICATION: The optimized chitosan/starch/garlic extract (Ch/De/GE) film developed in this study shows promising potential for application in the food packaging industry, particularly in extending the shelf life of perishable items like bell peppers. Its enhanced antibacterial properties, along with its ability to maintain visual appearance and reduce weight loss, make it an effective natural preservative that could replace synthetic additives in food packaging. By incorporating this biodegradable film into packaging solutions, producers can offer safer, more sustainable products that meet consumer demand for natural and environmentally friendly options.

Mechanisms of slow-release antibacterial properties in chitosan­titanium dioxide stabilized perilla essential oil Pickering emulsions: Focusing on oil-water interfacial behaviors.

Perilla essential oil (PLEO) offers benefits for food preservation and healthcare, yet its instability restricts its applications. In this study, chitosan (CS) and TiO2 used to prepare composite particles. TiO2, after being modified with sodium laurate (SL), was successfully introduced at 0.1 %-3 % into the CS matrix. The resulting CS-SL-TiO2 composite particles can be formed by intertwining and rearranging through intramolecular and intermolecular interactions, and form an O/W interface with stability and viscoelasticity. The Pickering emulsions stabilized by these particles exhibit non-Newtonian pseudoplastic behavior, shear-thinning properties, and slow-release characteristics, along with antibacterial activity. Emulsions with 0.5 % and 1 % CS-SL-TiO2 composites demonstrated superior antibacterial effects against Escherichia coli and Staphylococcus aureus. The study revealed that all emulsions undergo Fickian diffusion and a sustained release of PLEO, with the Ritger-Peppas model best describing this release mechanism. The slow-release behaviors positively correlates with interfacial pressure, composite particle size, composite particle potential, composite contact angle, emulsion particle size and emulsion potential, but negatively correlates with diffusion rate, penetration rate, release kinetics and release rate. The findings lay groundwork for developing slow-release antimicrobial emulsions within polysaccharide matrices, showcasing promise for antimicrobial packaging solutions and enhanced food preservation techniques.

Preparation methods, applications, toxicity and mechanisms of silver nanoparticles as bactericidal agent and superiority of green synthesis method.

Silver nanoparticles (SNPs) are a type of nanomaterial with wide applications in water treatment, medicine, food packaging, and industrial processes. Their unique optical, electrical, thermal conductivity, and biological properties distinguish them from other metal ions and liken them to noble metals like gold and copper. The present review explores the diverse applications, preparation techniques, mechanism of action of SNPs, and properties of SNPs focusing on their bactericidal activities and potential impacts on human health. Different preparation methods, encompassing chemical, physical, and biological techniques, were reviewed and analyzed to comprehend their effect on the properties and applications of SNPs. Studies revealed that the SNPs exhibit excellent antibactericidal properties. Mechanisms underlying their antimicrobial effects were explored, primarily focusing on pathogen-scavenging activities. Despite the promising benefits of SNPs, their potential toxicity to human health must be carefully managed. Regulatory standards, such as those set by WHO and USEPA; establish a maximum tolerable limit of 0.1 mg/L to mitigate health risks associated with SNP exposure. It is recommended to continue research into safer applications and alternative formulations of SNPs to minimize potential health risks while maximizing their beneficial applications across different industries.

A novel antimicrobial peptide WBp-1 from wheat bran: Purification, characterization and antibacterial potential against Listeria monocytogenes.

This study introduces a novel antimicrobial peptide (AMP), WBp-1, isolated from wheat bran and purified via reversed-phase high-performance liquid chromatography. The amino acid sequence, determined as IITGASSGIGKAIAKHFI by LC-MS/MS, was composed predominantly of alkaline and hydrophobic residues. WBp-1 was predicted to be a stable, hydrophobic, cationic peptide with an α-helical structure. Moreover, it displayed significant antibacterial efficacy against Listeria monocytogenes, with a minimum inhibitory concentration of 150 µg/mL. Further mechanistic studies suggest that WBp-1 exerts its bactericidal activity by disrupting cell membrane integrity, impeding peptidoglycan synthesis by binding to penicillin-binding protein 4 via hydrogen bonding, increasing cell permeability, altering membrane potential and fluidity, and altering surface hydrophobicity. Interestingly, WBp-1 showed minimal hemolytic activity and cytotoxicity against LO2 cells, even at 16× MIC. These findings highlight the strong potential of WBp-1 as a novel antibacterial agent and food preservative against Listeria monocytogenes.

Antibacterial Activity and Multi-Targeted Mechanism of Action of Suberanilic Acid Isolated from Pestalotiopsis trachycarpicola DCL44: An Endophytic Fungi from Ageratina adenophora.

Methicillin-resistant Staphylococcus aureus (MRSA) is a highly threatening foodborne pathogen capable of causing severe organ and life-threatening diseases. Over the past years, various commercial antibiotics have been used to treat MRSA infections. However, these commercial antibiotics have not yielded efficient results and also cause other side effects; therefore, there is a need for the development of effective alternatives to replace these commercial antibiotics. Suberanilic acid, an amide alkaloid obtained from the endophytic fungus Pestalotiopsis trachycarpicola DCL44, has been identified as a significant antimicrobial agent. However, its antibiotic properties on multi-drug-resistant bacteria such as MRSA have not been fully explored. Therefore, to investigate the potential antimicrobial mechanism of suberanilic acid against MRSA, a quantitative proteomics approach using tandem mass tagging (TMT) was used. The results obtained in the study revealed that suberanilic acid targets multiple pathways in MRSA, including disruption of ribosome synthesis, inhibition of membrane translocation for nutrient uptake (ABC transporter system), and causing dysregulation of carbohydrate and amino acid energy metabolism. These results provide new insights into the mechanism of action of suberanilic acid against MRSA and offer technical support and a theoretical basis for the development of novel food antimicrobial agents derived from endophytic fungal origin.

In Vitro Antimicrobial Synergistic Activity and the Mechanism of the Combination of Naringenin and Amikacin Against Antibiotic-Resistant Escherichia coli.

Bacterial drug resistance is becoming an increasingly serious problem, and the development of antibacterial synergists is urgently needed. Combining existing antibiotics with promising nonantibiotic agents is one strategy that has been shown to be effective at overcoming the widespread emergence of antibiotic-resistant pathogens. In this study, we investigated the antibacterial activities and mechanism of naringenin (NG) combined with amikacin (AMK) against multidrug-resistant Escherichia coli (E. coli). We first measured the fractional inhibitory concentration (FIC) of NG combined with antibiotics via the checkerboard method. The results indicated that the combination of NG and AMK had a synergistic effect on E. coli ATCC 25922 and E. coli C7F3. In addition, this synergistic effect was verified by time-kill assays. Moreover, scanning electron microscopy (SEM) was used to observe cell morphology. The results showed that the cell wall of E. coli was destroyed. Furthermore, we assessed the leakage of alkaline phosphatase (AKP), K+, and protein. The extracellular AKP activity increased after the combinational group of 1/2MIC NG and 1/2MIC AMK, suggesting an impairment in cell wall permeability. An increase in the leakage of intracellular K+ and protein indicated an increase in cell inner membrane permeability. These results revealed that NG and AMK inhibited E. coli by damaging cell walls and membranes. In addition, PI uptake rapidly increased after treatment with NG and AMK. Confocal laser scanning microscopy (CLSM) revealed that NG caused cell wall and cell membrane damage in E. coli. In summary, our results provide a new strategy for responding to the development of E. coli drug resistance.

Application of nano-ZnO in the food preservation industry: antibacterial mechanisms, influencing factors, intelligent packaging, preservation film and safety.

In recent years, how to improve the functional performance of food packaging materials has received increasing attention. One common inorganic material, nanometer zinc oxide (ZnO-NPs), has garnered significant attention due to its excellent antibacterial properties and sensitivity. Consequently, ZnO-NP-based functional packaging materials are rapidly developing in the food industry. However, there is currently a lack of comprehensive and systematic reviews on the use of ZnO-NPs as functional fillers in food packaging. In this review, we introduced the characteristics and antibacterial mechanism of ZnO-NPs, and paid attention to the factors affecting the antibacterial activity of ZnO-NPs. Furthermore, we systematically analyzed the application of intelligent packaging and antibacterial packaging containing ZnO-NPs in the food industry. At the same time, this paper also thoroughly investigated the impact of ZnO-NPs on various properties including thickness, moisture resistance, water vapor barrier, mechanical properties, optical properties, thermal properties and microstructure of food packaging materials. Finally, we discussed the migration and safety of ZnO-NPs in packaging materials. ZnO-NPs are safe and have negligible migration rates, simultaneously their sensitivity and antibacterial properties can be used to detect the quality changes of food during storage and extend its shelf life.

Macrolactin XY, a Macrolactin Antibiotic from Marine-Derived Bacillus subtilis sp. 18.

Two new compounds, macrolactin XY (1) and (5R, 9S, 10S)-5-(hydroxymethyl)-1,3,7-decatriene-9,10-diol (2), together with nine known compounds (3-11) were isolated from the marine Bacillus subtilis sp. 18 by the OSMAC strategy. These compounds were evaluated for antibacterial activity against six tested microorganisms. Compounds 1-5 and 7-10 showed varied antibacterial activity, with the minimum inhibitory concentration (MIC) ranging from 3 to 12 µg/mL. Macrolactin XY (1) was found to possess superior antibacterial activity, especially exhibiting significant effectiveness against Enterococcus faecalis. The antibacterial activity mechanism against E. faecalis was investigated. The mechanism may disrupt bacterial cell membrane integrity and permeability, and also inhibit the expression of genes associated with bacterial energy metabolism, as established by the experiments concerning cell membrane potential, SDS-PAGE electrophoresis, cell membrane integrity, and key gene expressions. This study offers valuable insights and serves as a theoretical foundation for the future development of macrolactins as antibacterial precursors.

Membrane Damage and Metabolic Disruption as the Mechanisms of Linalool against Pseudomonas fragi: An Amino Acid Metabolomics Study.

Pseudomonas fragi (P. fragi) is usually detected in low-temperature meat products, and seriously threatens food safety and human health. Therefore, the study investigated the antibacterial mechanism of linalool against P. fragi from membrane damage and metabolic disruption. Results from field-emission transmission electron microscopy (FETEM) and atomic force microscopy (AFM) showed that linalool damage membrane integrity increases surface shrinkage and roughness. According to Fourier transform infrared (FTIR) spectra results, the components in the membrane underwent significant changes, including nucleic acid leakage, carbohydrate production, protein denaturation and modification, and fatty acid content reduction. The data obtained from amino acid metabolomics indicated that linalool caused excessive synthesis and metabolism of specific amino acids, particularly tryptophan metabolism and arginine biosynthesis. The reduced activities of glucose 6-phosphate dehydrogenase (G6PDH), malate dehydrogenase (MDH), and phosphofructokinase (PFK) suggested that linalool impair the respiratory chain and energy metabolism. Meanwhile, genes encoding the above enzymes were differentially expressed, with pfkB overexpression and zwf and mqo downregulation. Furthermore, molecular docking revealed that linalool can interact with the amino acid residues of G6DPH, MDH and PFK through hydrogen bonds. Therefore, it is hypothesized that the mechanism of linalool against P. fragi may involve cell membrane damage (structure and morphology), disturbance of energy metabolism (TCA cycle, EMP and HMP pathway) and amino acid metabolism (cysteine, glutamic acid and citrulline). These findings contribute to the development of linalool as a promising antibacterial agent in response to the food security challenge.

Mining and characterization of novel antimicrobial peptides from the large-scale microbiome of Shanxi aged vinegar based on metagenomics, molecular dynamics simulations and mechanism validation.

The study aimed to mine and characterize novel antimicrobial peptides (AMPs) from the Shanxi aged vinegar microbiome. Utilizing machine learning techniques, AlphaFold2 structure prediction and molecular dynamics simulations, six novel AMPs were innovatively mined from 98,539 peptides based on metagenomic data, of which one peptide secreted by Lactobacillus (named La-AMP) was experimentally validated to have remarkable bactericidal effects against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) with high stability and no hemolytic activity. Scanning electron microscopy revealed that La-AMP caused irreversible damage to cell membranes of S. aureus and E. coli, a finding further confirmed by calcein-AM/propidium iodide staining. Additionally, La-AMP induced nucleic acid leakage and reactive oxygen species accumulation in bacterial cells. It was found to bind to DNA gyrase through salt bridges, hydrogen bonds, and hydrophobic interactions, ultimately inducing apoptosis. Thus, La-AMP exhibited encouraging promise as a valuable bioactive component for the development of natural preservatives.

Bioinspired construction of ATP/Co-Al-Zn LDH nanozyme with enhanced peroxidase-mimic performance for efficient bactericidal activity through membrane disruption.

In recent years, overuse of antibiotics has led to emerging antibiotic-resistant strains of bacteria. Consequently, creating new, highly productive antibacterial agents is crucial. In this work, we synthesized copper-aluminum-zinc layered double hydroxide (Co-Al-Zn LDH) and modified it using adenosine triphosphate. After characterization, the enzyme-like activity of the prepared particles was evaluated. The results indicated peroxidase-mimic performance of ATP/Co-Al-Zn LDH with Km values of 0.38 mM and 1.69 mM for TMB (3,3',5,5'-tetramethylbenzidine) and hydrogen peroxide (H2O2), respectively, which were lower than that of horseradish peroxidase. The highest peroxidase-like activity of ATP/Co-Al-Zn LDH was achieved at 20 °C, pH 4, with a 1.02 mg/mL catalyst, 231 µM TMB, and 1.9 mM H2O2. The bactericidal activity of the developed nanozyme was studied against E. coli and S. aureus. The peroxidase-mimic nanozyme decomposes H2O2 and generates free radicals to kill bacteria in vitro. The minimum inhibitory concentration (MIC) of ATP/Co-Al-Zn LDH was 15 µg/mL and 20 µg/mL for S. aureus and E. coli, respectively. The morphological characteristics of the nanozyme-treated bacterial cells showed dramatic changes in bacterial morphology. Our results revealed higher antibacterial activity of ATP/Co-Al-Zn LDH against S. aureus. Therefore, the developed nanozyme could serve as a substitute for conventional antibiotics.

Antibacterial activity and mechanism of chelerythrine against Streptococcus agalactiae.

Streptococcus agalactiae (S.agalactiae), also known as group B Streptococcus (GBS), is a highly infectious pathogen. Prolonged antibiotic usage leads to significant issues of antibiotic residue and resistance. Chelerythrine (CHE) is a naturally occurring benzophenidine alkaloid and chelerythrine chloride (CHEC) is its hydrochloride form with diverse biological and pharmacological activities. However, the antibacterial mechanism of CHEC against GBS remains unclear. Thus, this study aims to investigate the in vitro antibacterial activity of CHEC on GBS and elucidate its underlying mechanism. The antibacterial effect of CHEC on GBS was assessed using inhibitory zone, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) assays, as well as by constructing a time-kill curve. The antibacterial mechanism of CHEC was investigated through techniques such as scanning electron microscopy (SEM) and transmission electron microscopy (TEM), measurement of alkaline phosphatase (AKP) activity, determination of Na+ K+, Ca2+ Mg2+-adenosine triphosphate (ATP) activity, observation of membrane permeability, and analysis of intracellular reactive oxygen species (ROS) and mRNA expression levels of key virulence genes. The results demonstrated that the inhibition zone diameters of CHEC against GBS were 14.32 mm, 12.67 mm, and 10.76 mm at concentrations of 2 mg/mL, 1 mg/mL, and 0.5 mg/mL, respectively. The MIC and MBC values were determined as 256 µg/mL and 512 µg/mL correspondingly. In the time-kill curve, 8 × MIC, 4 × MIC and 2 × MIC CHEC could completely kill GBS within 24 h. SEM and TEM analyses revealed significant morphological alterations in GBS cells treated with CHEC including shrinkage, collapse, and leakage of cellular fluids. Furthermore, the antibacterial mechanism underlying CHEC's efficacy against GBS was attributed to its disruption of cell wall integrity as well as membrane permeability resulting in extracellular release of intracellular ATP, AKP, Na+ K+, Ca2+ Mg2+. Additionally CHEC could increase the ROS production leading to oxidative damage and downregulating mRNA expression levels of key virulence genes in GBS cells. In conclusion, CHEC holds potential as an antimicrobial agent against GBS and further investigations are necessary to elucidate additional molecular mechanisms.