Bacterial Membrane Permeability Research Articles

Self-Assembling Peptide Dendron Nanoparticles with High Stability and a Multimodal Antimicrobial Mechanism of Action.

Self-assembling nanometer-scale structured peptide polymers and peptide dendrimers have shown promise in biomedical applications due to their versatile properties and easy availability. Herein, self-assembling peptide dendron nanoparticles (SPDNs) with potent antimicrobial activity against a range of bacteria were developed based on the nanoscale self-assembly of an arginine-proline repeat branched peptide dendron bearing a hexadecanoic acid chain. The SPDNs are biocompatible, and our most active peptide dendron nanoparticle, C16-3RP, was found to have negligible toxicity after both in vitro and in vivo studies. Furthermore, the C16-3RP nanoparticles showed excellent stability under physiological concentrations of salt ions and against serum and protease degradation, resulting in highly effective treatment in a mouse acute peritonitis model. Comprehensive analyses using a series of biofluorescence, microscopy, and transcriptome sequencing techniques revealed that C16-3RP nanoparticles kill Gram-negative bacteria by increasing bacterial membrane permeability, inducing cytoplasmic membrane depolarization and drastic membrane disruption, inhibiting ribosome biogenesis, and influencing energy generation and other processes. Collectively, C16-3RP nanoparticles show promising biocompatibility and in vivo therapeutic efficacy without apparent resistance development. These advancements may facilitate the development of peptide-based antibiotics in clinical settings.

In-Cell FRET Indicates Magainin Peptide Induced Permeabilization of Bacterial Cell Membranes at Lower Peptide-to-Lipid Ratios Relevant to Liposomal Studies.

Antimicrobial peptides (AMPs) are promising candidates for anti-infective drugs. The majority of AMPs are considered to disrupt the lipid matrix of bacterial membranes, exerting bactericidal activity. A number of biophysical studies have been carried out to elucidate the underlying molecular mechanisms. However, the fact that the number of peptide molecules bound to a bacterial cell under bactericidal conditions is much larger than that expected from liposomal studies raises the question of whether membrane permeabilization mechanisms proposed by liposomal studies are relevant to bacteria. In this study, the peptide-to-lipid molar ratio needed for an antimicrobial magainin peptide to permeabilize the cell membrane of the Gram-positive bacterium Bacillus megaterium was estimated by random fluorescence resonance energy transfer from a BODIPY FL-labeled lipid to a Texas Red-labeled peptide. The comparison of the observed energy transfer efficiency with the two-dimensional energy transfer theory estimated that the leakage of the calcein dye from bacterial cells occurred at a peptide-to-lipid molar ratio of 0.025. At this ratio, the peptide induced dye leakage from liposomes mimicking the bacterial membrane, indicating that the lipid matrix is a target of membrane-acting AMPs and that liposomes are a useful model system to investigate their mechanisms of action. Furthermore, a binding assay suggested that most peptide molecules were bound to cellular components other than cell membranes.

Biochemical and Mutational Analysis of Radical S-Adenosyl-L-Methionine Adenosylhopane Synthase HpnH from Zymomonas mobilis Reveals that the Conserved Residue Cysteine-106 Reduces a Radical Intermediate and Determines the Stereochemistry.

Adenosylhopane is a crucial precursor of C35 hopanoids, which are believed to modulate the fluidity and permeability of bacterial cell membranes. Adenosylhopane is formed by a crosslinking reaction between diploptene and a 5'-deoxyadenosyl radical that is generated by the radical S-adenosyl-L-methionine (SAM) enzyme HpnH. We previously showed that HpnH from Streptomyces coelicolor A3(2) (ScHpnH) converts diploptene to (22R)-adenosylhopane. However, the mechanism of the stereoselective C-C bond formation was unclear. Thus, here, we performed biochemical and mutational analysis of another HpnH, from the ethanol-producing bacterium Zymomonas mobilis (ZmHpnH). Similar to ScHpnH, wild-type ZmHpnH afforded (22R)-adenosylhopane. Conserved cysteine and tyrosine residues were suggested as possible hydrogen sources to quench the putative radical reaction intermediate. A Cys106Ala mutant of ZmHpnH had one-fortieth the activity of the wild-type enzyme and yielded both (22R)- and (22S)-adenosylhopane along with some related byproducts. Radical trapping experiments with a spin-trapping agent supported the generation of a radical intermediate in the ZmHpnH-catalyzed reaction. We propose that the thiol of Cys106 stereoselectively reduces the radical intermediate generated at the C22 position by the addition of the 5'-deoxadenosyl radical to diploptene, to complete the reaction.

Antibacterial activity and mechanism of action of an ayurvedic formulation Khadirarishta

AimTo evaluate the antibacterial effect of Khadirarishta (An Ayurvedic traditional formulation) against some of the bacterial strains as well as to investigate the probable mechanism of action against sensitive bacterial strains by using standard procedures. MethodsThe antibacterial activity of Khadirarishta against some of the bacterial strains was evaluated by using bacterial susceptibility assay and the broth Microdilution method by using microplate reader and p-iodonitrotetrazolium chloride (INT) reagent assay. The mechanism of action of Khadirarishta was observed by its effects on the bacterial membrane followed by quantification of effluxes components. Resistance analysis and Combination effect of Khadirarishta was also performed to explore its antibacterial dimensions. HPLC analysis of Khadirarishta was carried out for identifying the different marker compounds. ResultsKhadirarishta revealed a significant (p < 0.05) antibacterial spectrum against E. coli, S. aureus, and S. enterica bacterial strains. The mechanism of action of Khadirarishta seems to be linked with the disruption of bacterial membrane permeability which leads to the smooth entry of active compounds, efflux of nucleotide material and K+ ions. The four major marker compounds viz., gallic acid, catechin, ellagic acid, and eugenol were identified by HPLC analysis. ConclusionThis study shows that Khadirarishta has antibacterial potency and has a vigorous effect on the bacterial cell wall and membrane permeability. This could be due to the presence of identified marker compounds in this herb-decoction based Ayurvedic traditional formulation.

Antimicrobial, Antibiofilm, and Anti-persister Activities of Penfluridol Against Staphylococcus aureus.

Staphylococcus aureus has increasingly attracted global attention as a major opportunistic human pathogen owing to the emergence of biofilms (BFs) and persisters that are known to increase its antibiotic resistance. However, there are still no effective antimicrobial agents in clinical settings. This study investigated the antimicrobial activity of penfluridol (PF), a long-acting antipsychotic drug, against S. aureus and its clinical isolates via drug repurposing. PF exhibited strong bactericidal activity against S. aureus, with a minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of 4–8 and 16–32 μg/ml, respectively. PF could significantly inhibit biofilm formation and eradicate 24 h preformed biofilms of S. aureus in a dose-dependent manner. Furthermore, PF could effectively kill methicillin-resistant S. aureus (MRSA) persister cells and demonstrated considerable efficacy in a mouse model of subcutaneous abscess, skin wound infection, and acute peritonitis caused by MRSA. Notably, PF exerted almost no hemolysis activity on human erythrocytes, with limited cytotoxicity and low tendency to cause resistance. Additionally, PF induced bacterial membrane permeability and ATP release and further caused membrane disruption, which may be the underlying antibacterial mechanism of PF. In summary, our findings suggest that PF has the potential to serve as a novel antimicrobial agent against S. aureus biofilm- or persister-related infections.

Dodecapeptide Cathelicidins of Cetartiodactyla: Structure, Mechanism of Antimicrobial Action, and Synergistic Interaction With Other Cathelicidins.

In this study, dodecapeptide cathelicidins were shown to be widespread antimicrobial peptides among the Cetruminantia clade. In particular, we investigated the dodecapeptide from the domestic goat Capra hircus, designated as ChDode and its unique ortholog from the sperm whale Physeter catodon (PcDode). ChDode contains two cysteine residues, while PcDode consists of two dodecapeptide building blocks and contains four cysteine residues. The recombinant analogs of the peptides were obtained by heterologous expression in Escherichia coli cells. The structures of the peptides were studied by circular dichroism (CD), FTIR, and NMR spectroscopy. It was demonstrated that PcDode adopts a β-hairpin structure in water and resembles β-hairpin antimicrobial peptides, while ChDode forms a β-structural antiparallel covalent dimer, stabilized by two intermonomer disulfide bonds. Both peptides reveal a significant right-handed twist about 200 degrees per 8 residues. In DPC micelles ChDode forms flat β-structural tetramers by antiparallel non-covalent association of the dimers. The tetramers incorporate into the micelles in transmembrane orientation. Incorporation into the micelles and dimerization significantly diminished the amplitude of backbone motions of ChDode at the picosecond-nanosecond timescale. When interacting with negatively charged membranes containing phosphatidylethanolamine (PE) and phosphatidylglycerol (PG), the ChDode peptide adopted similar oligomeric structure and was capable to form ion-conducting pores without membrane lysis. Despite modest antibacterial activity of ChDode, a considerable synergistic effect of this peptide in combination with another goat cathelicidin – the α-helical peptide ChMAP-28 was observed. This effect is based on an increase in permeability of bacterial membranes. In turn, this mechanism can lead to an increase in the efficiency of the combined action of the synergistic pair ChMAP-28 with the Pro-rich peptide mini-ChBac7.5Nα targeting the bacterial ribosome.

Investigation of the Antibacterial Activity and Efflux Pump Inhibitory Effect of Cycas thouarsii R.Br. Extract against Klebsiella pneumoniae Clinical Isolates.

The vast spread of multidrug-resistant bacteria has encouraged researchers to explore new antimicrobial compounds. This study aimed to investigate the phytochemistry and antibacterial activity of Cycas thouarsii R.Br. leaves extract against Klebsiella pneumoniae clinical isolates. The minimum inhibitory concentration (MIC) values of C. thouarsii extract ranged from 4 to 32 µg/mL. The impact of the treatment of the isolates with sub-inhibitory concentrations of C. thouarsii extract was investigated on the bacterial growth, membrane integrity, inner and outer membrane permeability, membrane depolarization, and bacterial morphology using a scanning electron microscope (SEM) and on the efflux activity using qRT-PCR. Interestingly, most K. pneumoniae isolates treated with C. thouarsii extract showed growth inhibition—a decrease in membrane integrity. In addition, we observed various morphological changes, a significant increase in inner and outer membrane permeability, a non-significant change in membrane depolarization, and a decrease in efflux activity after treatment. The phytochemical investigation of C. thouarsii extract revealed the isolation of one new biflavonoid, 5,7,7″,4‴-tetra-O-methyl-hinokiflavone (3), and five known compounds, stigmasterol (1), naringenin (2), 2,3-dihydrobilobetin (4), 4′,4‴-O-dimethyl amentoflavone (5), and hinokiflavone (6), for the first time. Moreover, the pure compounds′ MICs′ ranged from 0.25 to 2 µg/mL. Thus, C. thouarsii could be a potential source for new antimicrobials.

Antimicrobial Defensin and DNA Traps in Manila Clam Ruditapes philippinarum: Implications for Their Roles in Immune Responses

Although defensins have been isolated from a variety of metazoan, their role in cellular immunity has not been answered. In the study, we found that the hemocytes of the Manila clams Ruditapes philippinarum release defensin (designated as Rpdef3) in response to Vibrio parahaemolyticus challenge. The antimicrobial Rpdef3 was proved to be involved in the extracellular traps (ETs) that hemocytes released in response to Vibrio challenge. Scanning electron microscopy observation proved the patterns how ETs eliminate invading bacteria. Furthermore, Rpdef3 involved in ETs had broad-spectrum antimicrobial effect on both Gram-negative bacteria and Gram-positive bacteria. ELISA assay revealed that Rpdef3 could bind lipopolysaccharides and peptidoglycan in a dose-dependent manner. As concerned to the antibacterial mechanisms, Rpdef3 can cause bacterial membrane permeabilization, leading to cell death. As a result, Rpdef3 might contribute to the trap and the elimination of invading Vibrio in clam ETs. Taken together, our study suggest that the formation of ETs is a defense mechanism triggered by bacterial stimulation, coupled with antibacterial defensin.

Synergistic Photodynamic and Photothermal Antibacterial Activity of In Situ Grown Bacterial Cellulose/MoS2-Chitosan Nanocomposite Materials with Visible Light Illumination.

Owing to the rise in prevalence of multidrug-resistant pathogens attributed to the overuse of antibiotics, infectious diseases caused by the transmission of microbes from contaminated surfaces to new hosts are an ever-increasing threat to public health. Thus, novel materials that can stem this crisis, while also functioning via multiple antimicrobial mechanisms so that pathogens are unable to develop resistance to them, are in urgent need. Toward this goal, in this work, we developed in situ grown bacterial cellulose/MoS2-chitosan nanocomposite materials (termed BC/MoS2-CS) that utilize synergistic membrane disruption and photodynamic and photothermal antibacterial activities to achieve more efficient bactericidal activity. The BC/MoS2-CS nanocomposite exhibited excellent antibacterial efficacy, achieving 99.998% (4.7 log units) and 99.988% (3.9 log units) photoinactivation of Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus, respectively, under visible-light illumination (xenon lamp, 500 W, λ ≥ 420 nm, and 30 min). Mechanistic studies revealed that the use of cationic chitosan likely facilitated bacterial membrane disruption and/or permeability, with hyperthermia (photothermal) and reactive oxygen species (photodynamic) leading to synergistic pathogen inactivation upon visible-light illumination. No mammalian cell cytotoxicity was observed for the BC/MoS2-CS membrane, suggesting that such composite nanomaterials are attractive as functional materials for infection control applications.

In vitro activity of lactoferricin solution against Malassezia pachydermatis from otitis externa in dogs and cats.

Malassezia pachydermatis is a commensal organism of the skin, yet it may induce dermatitis and/or otitis. Lactoferricin (Lfcin) is an antimicrobial peptide obtained by the pepsin-mediated digestion of lactoferrin, a multifunctional innate-defence milk protein. The antibacterial activity of Lfcin is thought to cause alteration of bacterial membrane permeability, thus inducing cell death. The aim of this study was to evaluate in vitro antifungal activity of different dilutions of Lfcin solution against M.pachydermatis strains isolated from ears of dogs and cats with otitis externa. Fifty clinical Malassezia isolates from 40 dogs and 10 cats were tested. A dilution method in microtitre plates was used starting with a 20% Lfcin water solution. The different dilutions were 2:1 (13.3%), 1:1 (10%), 1:2 (6.7%), 1:5 (3.3%) and 1:10 (1.8%). Results were expressed as the value at which the growth of 50% (minimal fungicidal concentration MFC50 ) and 90% (MFC90 ) of yeast cells was inhibited. All strains showed susceptibility to 20% Lfcin solution (100%). With 1:5 and 1:10 dilutions corresponding to 3.3% and 1.8% Lfcin solutions, all strains showed resistance. The MFC50 and MFC90 values were observed at 13.3% and 20% Lfcin solutions. The results indicated that Lfcin solution exhibits the antimicrobial activity specific to antimicrobial peptides. In particular, the 20% solution can be effective in killing M.pachydermatis isolated strains. Some susceptibilities also are evident at lower concentrations of ≤6.7% for four strains.

Multidisciplinary Studies of Folk Medicine "Five Thieves' Oil" (Olejek Pięciu Złodziei) Components.

To meet the growing interest in natural antibacterial agents, we evaluated the physicochemical and biological properties of the folk medicine known as “five thieves’ oil” (Polish name: olejek pięciu złodziei). Five thieves’ oil consists of a mixture of five oils: rosemary, lemon, clove, eucalyptus, and cinnamon. In this study, we performed gas chromatography, FTIR, and UV–vis spectroscopic analysis, as well as L-a-b color tests, contact angle determination, and surface tension determination. To verify its antibacterial activity, the metabolic activity and changes in cell membrane permeability of bacteria of the genus Pseudomonas were studied. As a result, it was found that among the constituent oils, the oils of clove and cinnamon were the least volatile and, at the same time, had the strongest antibacterial activity. However, a mix of all the oils also showed comparable activity, which was even more pronounced for the oils after 4 weeks of aging. This effect can be linked to the high content of terpene derivatives such as eugenol and cinnamaldehyde, which can cause changes in bacterial membrane permeability, affecting cell activity and survival. This study is the first to characterize the constituents of the popular folk medicine five thieves’ oil, confirming and explaining its strong antibacterial activity, thus constituting a significant contribution to contemporary health education.

The Anti-Infectious Role of Sphingosine in Microbial Diseases.

Sphingolipids are important structural membrane components and, together with cholesterol, are often organized in lipid rafts, where they act as signaling molecules in many cellular functions. They play crucial roles in regulating pathobiological processes, such as cancer, inflammation, and infectious diseases. The bioactive metabolites ceramide, sphingosine-1-phosphate, and sphingosine have been shown to be involved in the pathogenesis of several microbes. In contrast to ceramide, which often promotes bacterial and viral infections (for instance, by mediating adhesion and internalization), sphingosine, which is released from ceramide by the activity of ceramidases, kills many bacterial, viral, and fungal pathogens. In particular, sphingosine is an important natural component of the defense against bacterial pathogens in the respiratory tract. Pathologically reduced sphingosine levels in cystic fibrosis airway epithelial cells are normalized by inhalation of sphingosine, and coating plastic implants with sphingosine prevents bacterial infections. Pretreatment of cells with exogenous sphingosine also prevents the viral spike protein of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) from interacting with host cell receptors and inhibits the propagation of herpes simplex virus type 1 (HSV-1) in macrophages. Recent examinations reveal that the bactericidal effect of sphingosine might be due to bacterial membrane permeabilization and the subsequent death of the bacteria.

Characterization of antibacterial activities and the related mechanisms of a β-defensin in turbot (Scophthalmus maximus)

β-defensins (BDs) are small-molecule cysteine-rich cationic antimicrobial peptides that play an important role in innate immunity against microbial invasion. In this study, a β-defensin (SmBD) was identified and characterized from turbot (Scophthalmus maximus). SmBD has an open reading frame containing 192 bp and encodes a protein of 63 amino acids. It has a predicted signal peptide consisting of 20 amino acids and a mature peptide with six conserved cysteine residues forming three disulfide bonds (C1- C5, C2-C4 and C3-C6). The 3D structure prediction shows an N-terminal α-helix and three reverse parallel β-folds. SmBD showed high sequence identity (70.45% - 95.24%) with β-defensin 2 group members from other teleost species. SmBD had the highest expression in skin of healthy turbot and was significantly up-regulated in mucosal tissues (skin, gill, intestine) after bacteria challenge. SmBD showed antibacterial activity against Vibrio anguillarum, Edwardsiella tarda, Escherichia coli and Streptococcus iniae, and inhibitory effect on the formation of bacterial biofilms. SmBD changed bacterial membrane permeability and affected bacterial morphology, as shown in fluorescence microscopy and scanning electron microscopy analyses, respectively, which indicated that SmBD might exercise antibacterial behaviors via “membrane-attack” mechanism.

Antibacterial activity and antibacterial mechanism of ethanol extracts of Lentinula edodes (Shiitake) and Agaricus bisporus (button mushroom)

ABSTRACT The aim of the present work was the comparison of antimicrobial activity, mechanism and components of the ethanol extract (EE) from Lentinula edodes Berk (Pegler) and Agaricus bisporus Sing (Lange). The main component of EE of A. bisporus was dianhydromannitol (20.1%), while isosorbide/dianhydromannitol (21.8%) was detected at a high rate in L. edodes ethanol extract by GC-MS . The common phenolic acids were determined as chlorogenic acid, syringic acid, rutin, p-coumaric acid, ferulic acid, 2-hydroxy cinnamic acid, protocatechuic acid, abscisic acid, and trans-cinnamic acid in both mushroom extract by HPLC-MWD . The MICs (minimum inhibitory concentration) of L. edodes EE on Klebsiella pneumoniae, Staphylococcus aureus, Enterococcus faecalis and Acinetobacter baumannii were between 5.1 mg ml−1 and 6.01 mg ml−1, while MICs of A. bisporus EE on the pathogens were between 5.8 mg ml−1 and 9.54 mg ml−1. The highest DRA decrease was in E. faecalis (69.1%) for L. edodes and S. aureus (71.0%) for A. bisporus in the 20th minute. As a result, L. edodes and A. bisporus have a similar antibacterial effect on the pathogens, and this inhibition effect caused DNA, protein leakage and destruction of permeability of bacterial cell membrane by bioactive molecules in mushroom extract.

Advances in the research of enterobacterial common antigen

The enterobacterial common antigen (ECA) is a polysaccharide composed of polysaccharide repeats that are located in the outer membrane of almost all Enterobacteriaceae bacteria and has diverse biological functions. ECA is synthesized by the synergistic action of multiple genes that are present in clusters on the genome of Enterobacteriaceae bacteria, forming the ECA antigen gene cluster, an important virulence factor that plays a role in host invasion and survival of Enterobacteriaceae in vivo. ECA also plays an important role in the maintenance of the bacterial outer membrane permeability barrier, flagella gene expression, swarming motility, and bile salts resistance. In addition, ECALPS, anchored in the core region of bacterial lipopolysaccharide, is an important surface antigen for bacteria, stimulating high levels of antibody production in the host and could be a target for vaccine research. This review summarizes ECA purification, genes involved in ECA biosynthesis, its immunological characteristics, biological functions and clinical applications.

Synergistic inactivation of bacteria based on a combination of low frequency, low-intensity ultrasound and a food grade antioxidant

This study evaluated a synergistic antimicrobial treatment using a combination of low frequency and a low-intensity ultrasound (LFU) and a food-grade antioxidant, propyl gallate (PG), against a model gram-positive (Listeria innocua) and the gram-negative bacteria (Escherichia coli O157:H7). Bacterial inactivation kinetic measurements were complemented by characterization of biophysical changes in liposomes, changes in bacterial membrane permeability, morphological changes in bacterial cells, and intracellular oxidative stress upon treatment with PG, LFU, and a combination of PG + LFU. Combination of PG + LFU significantly (>4 log CFU/mL, P < 0.05) enhanced the inactivation of both L. innocua and E. coli O157:H7 compared to PG or LFU treatment. As expected, L. innocua had a significantly higher resistance to inactivation compared to E. coli using a combination of PG + LFU. Biophysical measurements in liposomes, bacterial permeability measurements, and scanning electron microscope (SEM)-based morphological measurements show rapid interactions of PG with membranes. Upon extended treatment of cells with PG + LFU, a significant increase in membrane damage was observed compared to PG or LFU alone. A lack of change in the intracellular thiol content following the combined treatment and limited effectiveness of exogenously added antioxidants in attenuating the synergistic antimicrobial action demonstrated that oxidative stress was not a leading mechanism responsible for the synergistic inactivation by PG + LFU. Overall, the study illustrates synergistic inactivation of bacteria using a combination of PG + LFU based on enhanced membrane damage and its potential for applications in the food and environmental systems.

Co-effect of cadmium and iron oxide nanoparticles on plasmid-mediated conjugative transfer of antibiotic resistance genes

Conjunctive transfer of antibiotic resistance genes (ARGs) among bacteria driven by plasmids facilitated the evolution and spread of antibiotic resistance. Heavy metal exposure accelerated the plasmid-mediated conjunctive transfer of ARGs. Nanomaterials are well-known adsorbents for heavy metals removal, with the capability of combatting resistant bacteria/facilitating conjunctive transfer of ARGs. However, co-effect of heavy metals and nanomaterials on plasmid-mediated conjunctive transfer of ARGs was still unknown. In this study, we investigated the effect of the simultaneous exposure of Cd2+ and nano Fe2O3 on conjugative transfer of plasmid RP4 from Pseudomonas putida KT2442 to water microbial community. The permeability of bacterial cell membranes, antioxidant enzyme activities and conjugation gene expression were also investigated. The results suggested that the combination of Cd2+ and high concentration nano Fe2O3 (10 mg/L and 100 mg/L) significantly increased conjugative transfer frequencies of RP4 plasmid (p < 0.05). The most transconjugants were detected in the treatment of co-exposure to Cd2+ and nano Fe2O3, the majority of which were identified to be human pathogens. The mechanisms of the exacerbated conjugative transfer of ARGs were involved in the enhancement of cell membrane permeability, antioxidant enzyme activities, and mRNA expression levels of the conjugation genes by the co-effect of Cd2+ and nano Fe2O3. This study confirmed that the simultaneous exposure to Cd2+and nano Fe2O3 exerted a synergetic co-effect on plasmid-mediated conjunctive transfer of ARGs, emphasizing that the co-effect of nanomaterials and heavy metals should be prudently evaluated when combating antibiotic resistance.

Identification of CO2 induces oxidative stress to change bacterial surface properties

The surface properties of bacteria play an essential role in their abilities to perform transmembrane communication, adherence, immobilization, flocculation, etc. However, the responsiveness of bacterial surfaces to elevated atmospheric CO2 remains unknown. In this study, using the model bacteria, Paracoccus denitrificans, the effect of CO2 on the primary bacterial surface properties, specifically hydrophobicity and surface charge, has been explored. We found that hydrophilicity and negative surface charge both rose in conjunction with increased atmospheric CO2 concentrations. Studies of the potential mechanisms involved have illustrated that elevated CO2 significantly increases the production of polysaccharides in extracellular polymeric substances (EPS). Various hydrophilic groups and negative charges in these polysaccharides prompt hydrophilicity and surface charge variations in bacteria. Further research has identified that elevations in CO2 result in the accumulation of reactive species, specifically reactive nitrogen species (RNS). In this study, it was found that RNS damaged the permeability of bacterial membranes by inducing lipid peroxidation and then caused the leakage of intracellular substrate, which ultimately led to an increase in EPS polysaccharides. Our findings suggest that changes in bacterial surface properties due to atmospheric CO2 elevation, as well as the reactions these trigger, merit widespread attention.

Chemical composition, antibacterial activity and study of the interaction mechanisms of the main compounds present in the Alpinia galanga rhizomes essential oil

In this research, the antibacterial activity and the interaction mechanisms of the major chemical constituent present in the Alpinia galanga rhizomes essential oil (AGREO) were investigated. Ethyl cinnamate, methyl cinnamate and n-pentadecane were the major components of AGREO. The antibacterial mechanism of AGREO on Enterohemorrhagic Escherichia coli O157:H7 (EHEC O157) was evaluated and the results proved that the potent antibacterial activity of AGREO was due to higher passive permeability of bacterial cell membrane, followed by crucial intracellular components efflux. Additionally, AGREO disrupted the intracellular physiological metabolism of EHEC O157, including inhibition of P-type ATPases activity and down-regulation expression of four virulence genes associated with EHEC infections. Notably, molecular docking studies suggested that ethyl cinnamate probably competed with the structural NAD(P)+ for the binding sites of glucose-6-phosphate dehydrogenase (G6PD), resulting in a decrease in G6PD activity with subsequent suppression of respiratory metabolism. The spectroscopic techniques and molecular docking studies proved that the AGREO major components were prone to bind to the minor groove of DNA.

Making the Enterobacterial Common Antigen Glycan and Measuring Its Substrate Sequestration.

The enterobacterial common antigen (ECA), a three-sugar repeat unit polysaccharide produced by Enterobacteriaceae family members, impacts bacterial outer membrane permeability, and its biosynthesis affects the glycan landscape of the organism. ECA synthesis impacts the production of other polysaccharides by reducing the availability of shared substrates, the most notable of which is the 55-carbon polyisoprenoid bactoprenyl phosphate (BP), which serves as a carrier for the production of numerous bacterial glycans including ECA, peptidoglycan, O-antigen, and more. Here, using a combination of in vitro enzymatic synthesis and liquid chromatography-mass spectrometry (LC-MS) analysis of bacterial lysates, we provide biochemical evidence for the effect on endogenous polyisoprenoid pools from cell culture that arises from glycan pathway disruption. In this work, we have cloned and expressed each gene involved in ECA repeat unit biosynthesis and reconstituted the pathway in vitro, providing LC-MS characterized standards for the investigation of cellular glycan-linked intermediates and BP. We then generated ECA deficient mutants in genes associated with production of the polysaccharide, which we suspected would accumulate materials identical to our standards. We found that indeed accumulated products from these cells were indistinguishable from our enzymatically prepared standards, and moreover we observed a concomitant decrease in cellular BP levels with each mutant. This work provides the first direct biochemical evidence for the sequestration of BP upon the genetic disruption of glycan biosynthesis pathways in bacteria. This work also provides methods for the direct assessment of both the ECA glycan, and a new understanding of the dynamic interdependence of the bacterial polysaccharide repertoire.