RESUMO
Cobalt-free (Co-free) and nickel-rich (Ni-rich) cathode materials have attracted significant attention and undergone extensive studies due to their affordability and superior energy density. However, the commercialization of these Co-free materials is hindered by challenges such as cation disorder, irreversible phase changes, and inadequate high-voltage performance. To overcome these challenges, a Co-free ternary cathode material of Mg/Al double-pillared LiNiO2 (NMA) synthesized via a wet-coating and lithiation-sintering technique is proposed. Fundamental studies reveal that Mg and Al have the potential to form a distinctive double-pillar structure within the layered cathode, enhancing its structural stability. To be specific, the strategic placement of Mg and Al in Li and Ni layers, respectively, effectively reduces Li+/Ni2+ disorder and prevents irreversible phase transitions. Additionally, the inclusion of Mg and Al refines the primary grains and compacts the secondary grains in the cathode material, reducing stress from cyclic usage and preventing material cracking, thereby mitigating electrolyte erosion. As a result, NMA demonstrates exceptional electrochemical performance under a high charge cutoff voltage of 4.6 V. It maintains 70% of initial specific capacity after 500 cycles at 1 C and exhibits excellent rate performance, with a capacity of 162 mAh g-1 at 5 C and 149 mAh g-1 at 10 C. As a whole, the produced NMA achieves a high structural stability in cases of excessive delithiation, providing a groundbreaking solution for the development of cost-effective and high-energy-density cathode materials for lithium-ion batteries.
RESUMO
Nitric oxide (NO), produced through the denitrification pathway, regulates biofilm dynamics through the quorum sensing system in Pseudomonas aeruginosa. NO stimulates P. aeruginosa biofilm dispersal by enhancing phosphodiesterase activity to decrease cyclic di-GMP levels. In a chronic skin wound model containing a mature biofilm, the gene expression of nirS, encoding nitrite reductase to produce NO, was low, leading to reduced intracellular NO levels. Although low-dose NO induces biofilm dispersion, it is unknown whether it influences the formation of P. aeruginosa biofilms in chronic skin wounds. In this study, a P. aeruginosa PAO1 strain with overexpressed nirS was established to investigate NO effects on P. aeruginosa biofilm formation in an ex vivo chronic skin wound model and unravel the underlying molecular mechanisms. Elevated intracellular NO levels altered the biofilm structure in the wound model by inhibiting the expression of quorum sensing-related genes, which was different from an in vitro model. In Caenorhabditis elegans as a slow-killing infection model, elevated intracellular NO levels increased worms' lifespan by 18%. Worms that fed on the nirS-overexpressed PAO1 strain for 4 h had complete tissue, whereas worms that fed on empty plasmid-containing PAO1 had biofilms on their body, causing severe damage to the head and tail. Thus, elevated intracellular NO levels can inhibit P. aeruginosa biofilm growth in chronic skin wounds and reduce pathogenicity to the host. Targeting NO is a potential approach to control biofilm growth in chronic skin wounds wherein P. aeruginosa biofilms are a persistent problem.
Assuntos
Infecções por Pseudomonas , Pseudomonas aeruginosa , Animais , Pseudomonas aeruginosa/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico/farmacologia , Biofilmes , Percepção de Quorum , Virulência , Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/microbiologiaRESUMO
This study investigated the antivirulence capacity and mechanism of apple-skin-derived phloretin against Serratia marcescens NJ01, a vegetable spoilage bacterium. At 0.5 to 2 mg/mL doses, phloretin considerably inhibited the secretion of acyl homoserine lactones (AHLs), indicating that phloretin disrupted quorum sensing (QS) in S. marcescens NJ01. The dysfunction of QS resulted in reduced biofilms and the decreased production of protease, prodigiosin, extracellular polysaccharides (EPSs), and swimming and swarming motilities. Dysfunctional QS also weakened the activity of antioxidant enzymes and improved oxidative injury. The improved oxidative injury changed the composition of the membrane, improved membrane permeability, and eventually increased the susceptibility of biofilm cells to amikacin, netilmicin, and imipenem. The disrupted QS and enhanced oxidative stress also caused disorders of amino acid metabolism, energy metabolism, and nucleic acid metabolism, and ultimately attenuated the ability of S. marcescens NJ01 to induce spoilage. Our results indicated that phloretin can act as a potent drug to defend against spoilage by S. marcescens.
Assuntos
Percepção de Quorum , Serratia marcescens , Serratia marcescens/metabolismo , Antibacterianos/farmacologia , Antibacterianos/metabolismo , Biofilmes , Prodigiosina/farmacologiaRESUMO
Listeria monocytogenes, a foodborne pathogen that causes listeriosis with high fatality rate, exhibits multidrug resistance (MDR) known to be progressively increasing. Alternative antibacterial strategies are in high demand for treating this well-known pathogen. Anti-biofilm and anti-virulence strategies are being explored as novel approaches to treat bacterial infections. In this study, one rare antibacterial named setomimycin was isolated from Streptomyces cyaneochromogenes, which showed potent antibacterial activity against L. monocytogenes. Next, the inhibition of biofilm formation and listeriolysin O (LLO) production against L. monocytogenes were investigated at sub-minimal inhibitory concentrations (sub-MICs) of setomimycin alone or combined with kanamycin and amikacin. Crystal violet staining confirmed that setomimycin combining with kanamycin or amikacin could dramatically reduce biofilm formation against L. monocytogenes at sub-MICs, which was further evaluated by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). In the meantime, sub-MICs of setomimycin could significantly suppress the secretion of LLO. Furthermore, the transcription of genes associated with biofilms and main virulence factors, such as LLO, flagellum, and metalloprotease, were suppressed by setomimycin at sub-MICs. Hence, the study provided a deep insight into setomimycin as an alternative antibacterial agent against L. monocytogenes.
Assuntos
Listeria monocytogenes , Listeriose , Humanos , Amicacina/farmacologia , Canamicina/farmacologia , Listeriose/microbiologia , Biofilmes , Antibacterianos/farmacologia , Proteínas Hemolisinas/genéticaRESUMO
IMPORTANCE: Our previous study demonstrated that the expression of lapA was induced under phosphate depletion conditions, but its roles in virulence and biofilm formation by Pseudomonas aeruginosa remain largely unknown. This study presents a systematic investigation of the roles of lapA in virulence induction and biofilm formation by constructing a lapA-deficient strain with P. aeruginosa PAO1. The results showed that deletion of the lapA gene evidently reduced elastase activity, swimming motility, C4-HSL, and 3-oxo-C12-HSL production, and increased rhamnolipid production under phosphate depletion stress. Moreover, lapA gene deletion inhibited PAO1 biofilm formation in porcine skin explants by reducing the expression levels of las and rhl quorum sensing systems and extracellular polymeric substance synthesis. Finally, lapA gene deletion also reduced the virulence of PAO1 in Caenorhabditis elegans in fast-kill and slow-kill infection assays. This study provides insights into the roles of lapA in modulating P. aeruginosa virulence and biofilm formation under phosphate depletion stress.
Assuntos
Infecções por Pseudomonas , Percepção de Quorum , Humanos , Virulência , Pseudomonas aeruginosa , Biofilmes , Fosfatase Alcalina/farmacologia , Matriz Extracelular de Substâncias Poliméricas/metabolismo , Fosfatos/farmacologia , Fatores de Virulência/genética , Fatores de Virulência/metabolismo , Corantes , Antibacterianos/farmacologia , Proteínas de Bactérias/genéticaRESUMO
Abundant starch was isolated from Dioscorea zingiberensis C.H. Wright, a novel and underutilized industrial crop resource. In this study, an intelligent packaging film able to indicate food freshness was developed and characterized. D. zingiberensis starch (DZS) was bleached first, and its particle size, total starch content, amylose content, and gelatinization temperature were then measured. Butterfly pea (Clitoria ternatea Linn.) flowers were selected as the source of polyphenols, which rendered the prepared film intelligent and progressively blue-violet. SEM and FT-IR analyses showed the homogeneous dispersion of butterfly pea flower extract (BPE) in the film. The BPE-loaded film showed improved flexibility and resistance to UV and oxidation while maintaining sufficient mechanical strength and physical properties. Moreover, the film underwent a distinguishable color change from red to blue-violet and finally to green-yellow with increasing pH from 2 to 13. Similar color alteration also occurred when the film was exposed to ammonia. When the film was used to monitor the freshness of chicken stored at room temperature, it exhibited an obvious color change, implying its deterioration. Therefore, the newly developed BPE-DZS film, which was produced from readily accessible natural substances, can serve as an intelligent packaging material, indicating food freshness and prolonging shelf life.
Assuntos
Dioscorea , Amido , Amido/química , Antocianinas/química , Espectroscopia de Infravermelho com Transformada de Fourier , Embalagem de Alimentos , Carne , Concentração de Íons de HidrogênioRESUMO
Prosapogenin A is a secondary saponin in Dioscorea zingiberensis, and it showed remarkable pharmacological effects. Due to very low content and lack of well-developed biotransformation, its preparation was not efficient and clean. This study aims to establish an eco-friendly strategy for preparation of Prosapogenin A from plant material. Physical separation was employed to recycle starch and cellulose, and then D101 resin and polyamide packed-bed column was incorporated for purification of total steroidal saponins (TSS). After these pretreatments, purity of TSS was largely increased to 83.2% with recovery at 87.6%, which was subjected to enzymatic hydrolysis. Optimized reaction system was constructed in 0.20 M HAc-NaAc buffer (pH4.2) containing cellulase/TSS (3:1, w/w), and the hydrolysis was performed at 53 °C for 6 h. Consequently, TSS was almost completely hydrolyzed to Prosapogenin A, while the highest yield reached 5.62%. The newly proposed approach is promising for efficient preparation of Prosapogenin A in industrial applications.
Assuntos
Dioscorea , Saponinas , Hidrólise , Saponinas/farmacologia , BiotransformaçãoRESUMO
AIMS: In the current study the anti-virulence and anti-biofilm activities of the cinnamic acid derivative, 3-methoxycinnamic acid, was investigated against Agrobacterium tumefaciens. METHODS AND RESULTS: Based on the disc diffusion test and ß-galactosidase activity assay, 3-methoxycinnamic acid was shown to interfere with the quorum sensing (QS) system of A. tumefaciens. Crystal violet staining assay, phenol-sulfuric acid method, Bradford protein assay and confocal laser scanning microscopy (CLSM) revealed that the biofilm formation of A. tumefaciens was inhibited after the treatment of 3-methoxycinnamic acid. Employing high-performance liquid chromatography (HPLC) analysis of culture supernatant revealed that the production of 3-oxo-octanoylhomoserine lactone (3-oxo-C8-HSL) decreased concentration-dependently after treatment with 3-methoxycinnamic acid. Swimming and chemotaxis assays also indicated that 3-methoxycinnamic acid had a good effect on reducing the motility and chemotaxis of A. tumefaciens. In addition, the RT-qPCR, molecular docking and simulations further demonstrated that 3-methoxycinnamic acid could competitively inhibit the binding of 3-oxo-C8-HSL to TraR and down-regulate virulence-related genes. CONCLUSIONS: 3-Methoxycinnamic acid is proved to have good anti-virulence and anti-biofilm activities against A. tumefaciens. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study that investigates the anti-virulence and anti-biofilm activities of 3-methoxycinnamic acid against A. tumefaciens. With its potential QS-related virulence and biofilm inhibitory activities, 3-methoxycinnamic acid is expected to be developed as a potent pesticide or adjuvant for the prevention and treatment of crown gall caused by A. tumefaciens.
Assuntos
Agrobacterium tumefaciens , Praguicidas , Agrobacterium tumefaciens/metabolismo , Simulação de Acoplamento Molecular , Violeta Genciana/metabolismo , Violeta Genciana/farmacologia , Percepção de Quorum , Biofilmes , 4-Butirolactona , Fenóis/farmacologia , Praguicidas/farmacologia , beta-Galactosidase/metabolismoRESUMO
In this study, abundant starch was separated from the industrial crop Dioscorea zingiberensis C.H. Wright (DZW), and a novel bioactive packaging film loaded with oregano essential oil (OEO) was prepared and characterized. NaClO solution worked as a bleacher to prepare uniform starch powder from DZW tubers. OEO was selected from among three essential oils of Labiatae family plants for its strongest antibacterial activity. After the addition of OEO into the starch-based film, the UV-vis shielding property and antioxidant activity were enhanced. Meanwhile, the films still have a considerable performance in transparency, mechanical strength and water vapor permeability after incorporated with OEO. Furthermore, the 3% OEO-loaded starch film exhibited the strongest antibacterial activity against Bacillus subtilis, Escherichia coli and Staphylococcus aureus. It effectively lowered the total viable count of fresh chicken under 4 °C preservation conditions. These results revealed that the OEO-loaded DZW starch film can exert a positive effect on maintaining the quality and extending the shelf life of fresh meat. Therefore, readily accessible DZW tubers and oregano are very promising resources for application in degradable bioactive packaging film.
Assuntos
Dioscorea , Óleos Voláteis , Origanum , Animais , Antibacterianos/farmacologia , Antioxidantes/farmacologia , Galinhas , Escherichia coli , Embalagem de Alimentos/métodos , Óleos Voláteis/farmacologia , AmidoRESUMO
The natural product 4-hydroxycinnamic acid (HA) was firstly isolated from the metabolites of Phomopsis liquidambari, one endophytic fungus from Punica granatum leaves. The anti-QS potential of HA was evaluated by ß-galactosidase assay and acylated homoserine lactones (AHL) analysis. The MIC of HA was > 1.20 mM. Exposure to HA at sub-MIC concentrations (0.30-0.60 mM) remarkably reduced the ß-galactosidase activity and AHL secretion. Transcriptional analysis by qRT-PCR and docking simulation indicated that HA functions as an anti-QS agent by inhibiting the transcriptional levels of traI and traR rather than signal mimicry. The blocked QS lead to suppressed biofilm formation, motilities, and flagella formation after exposure to HA at concentrations ranging from 0.30 to 0.80 mM. The dysfunctional QS also resulted in repressed antioxidant enzymes and intensified oxidative stress. The intensified oxidative stress destroyed membrane integrity, induced energy supply deficiency, resulted in disorder of protein and nuclear acid metabolism, and ultimately weakened pathogenicity of A. tumefaciens. HA may have promising potential for controlling A. tumefaciens.
RESUMO
Pseudomonas aeruginosa is one of the most important foodborne pathogens that can persist in leafy green vegetables and subsequently produce biofilms. In this study, the synergistic effect of thymoquinone and nisin in reducing biofilm formation of P. aeruginosa on lettuce was evaluated, and their anti-virulence and anti-biofilm mechanisms were also investigated. At concentrations ranging from 0.5 to 2 mg/ml, thymoquinone inhibited the production of autoinducers and virulence factors, and enhanced the susceptibility of P. aeruginosa biofilms to nisin as evidenced by the scanning electron microscopy and confocal laser scanning microscopy. Integrated transcriptomics, metabolomics, and docking analyses indicated that thymoquinone treatment disrupted the quorum sensing (QS) system, altered cell membrane component, and down-regulated the expressions of genes related to virulence, efflux pump, and antioxidation. The changed membrane component and repressed efflux pump system enhanced membrane permeability and facilitated the entrance of nisin into cells, thus improving the susceptibility of biofilms to nisin. The dysfunctional QS and repressed antioxidant enzymes lead to the enhancement of oxidative stress. The enhanced oxidative stress disrupted energy metabolism and protein metabolism and ultimately attenuated the virulence and pathogenicity of P. aeruginosa PAO1. Our study indicated that thymoquinone has the potential to function as a QS-based agent to defend against foodborne pathogens in combination with nisin.
RESUMO
Ni2+/Ni4+ and O2-/On2- redoxs endow the Li-rich layered oxide of Li1.2Mn0.6Ni0.2O2 (LMNO) with a considerable specific capacity and higher voltage. However, during the repeated de-/lithiation, the constant structure degradation initiated from transition metal ion dissolvement and oxygen escape leads to rapid capacity decay, which severely hinders the commercial application of LMNO. Herein, Nb2O5 and LiNbO3 are fabricated on the outside of the LMNO substrate. With the appropriate ion radius, a small amount of Nb5+ enters the substrate, which could enlarge the crystal spacing and facilitate the fast Li+ transfer and, more importantly, change the valence state of Mn and induce the formation a Fd3Ì m transition phase on the interface between the coating layer and the interior LMNO. Density functional theory (DFT) calculation has proven that the transition phase could build double-way chemical bonds both inside and outside, and the LiNbO3 coated LMNO composite (LMNO@LNO) possesses a more stable and harmonious interface due to the higher bonding strength between LiNbO3 and the transition phase. Therefore, LMNO@LNO demonstrates the most outstanding rate capability and long-tern cycling stability (decay rate of 0.041% per cycle during 1000 cycling at 5 C). This work provides a new inspiration for the coating materials selection and the interface stability research for the LMNO cathodes.
RESUMO
Bacterial biofilms formation is one of the major reasons for treatment failure in chronic wound infections. Therefore, diagnostic biomarkers remain the best option for prevention and treatment of chronic wound infections by biofilms. Herein, Pseudomonas aeruginosa PAO1 was used to mimic biofilm development in porcine skin explants wells as ex vivo wound model. The microscopic imaging showed that PAO1 in porcine skin explants wells formed micro-colonies at 24 h, developed mushroom-like structure at 48 h, and at 72 h mushroom-like structure disappeared, remaining a thin bacterial lawn. RNA-seq data analysis revealed that the expression levels of genes involved in the type II hxc secretion system were significantly higher in biofilms than in planktonic cells, especially the expression of lapA encoding alkaline phosphatase. However, the expression levels of genes associated with denitrification pathway were markedly decreased in biofilms, especially the transcription of nirS encoding nitrite reductase to produce nitric oxide (NO). Therefore, their expressions and products were further detected using RT-qPCR and biochemical assays, respectively. The results found that the expression of lapA and alkaline phosphatase activity were induced, but the expression of nirS and intracellular NO were reduced at the whole biofilms cycle. The study indicates that LapA and NO would play an important role for P. aeruginosa biofilm formation in chronic wound infections. LapA would serve as potential target to monitor chronic wound infections by P. aeruginosa biofilms. Inducing NO would be used to treat chronic wound infections due to P. aeruginosa biofilms.
RESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Spleen-Yang deficiency (SYD) is one of the primary causes of many digestive diseases, such as ulcerative colitis (UC), and irritable bowel syndrome (IBS), but its endogenous metabolic characteristics are still unclear. Fuzi Lizhong pill (FLZP) is well-known for its powerful capacity for treating SYD; however, its mechanisms require further study. AIM OF THE STUDY: Herein, our present study aimed to investigate the essence of SYD from the perspective of metabolomics, and tried to reveal the anti-SYD action mechanisms of FLZP. MATERIALS AND METHODS: Firstly, the compound factor modeling method with the principle of "indiscipline in diet + excessive fatigue + intragastric administration of Senna water extracts" was used to establish Sprague Dawley (SD) rats as SYD model. Then, the visceral index, motilin (MTL), malonaldehyde (MDA), Interleukin 1 alpha (IL-1α), and Interleukin 6 (IL-6) levels were used to verify the anti-SYD effect of FLZP. In addition, serum samples were analyzed by UPLC-QE/MS metabolomics technique. Finally, the metabolic pathways associated with specific biomarkers were analyzed to research the possible mechanism underlying the action of FLZP. RESULTS: The expression of MTL, MDA, IL-1α, and IL-6 were regulated by FLZP, which suggested that it has relieved diarrhea and gastrointestinal motility disorder caused by SYD and had an anti-peroxidation, anti-inflammatory, and immune regulation effect. A total of 75 metabolites were found to be the potential biomarkers of SYD. Moreover, FLZP regulates 21 metabolites and 10 vital pathways including the tricarboxylic acid (TCA) cycle, sphingolipid metabolism, and histidine metabolism. CONCLUSION: SYD primarily causes disorders of amino acid metabolism, lipid metabolism, carbohydrate metabolism, metabolism of cofactors and vitamins, nucleotide metabolism, and translation. In addition, FLZP regulated carbohydrate, lipid, and amino acid metabolisms, gastrointestinal motility, digestive juice secretion, immune regulation, as well as antioxidant effects. Hence, FLZP had a good therapeutic effect on treatment of SYD. It might be a promising therapeutic agent for the treatment of SYD-related diseases.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Esplenopatias/tratamento farmacológico , Deficiência da Energia Yang/tratamento farmacológico , Animais , Biomarcadores/metabolismo , Modelos Animais de Doenças , Metabolismo dos Lipídeos/efeitos dos fármacos , Masculino , Metabolômica , Ratos , Ratos Sprague-Dawley , Esplenopatias/metabolismo , Síndrome , Deficiência da Energia Yang/metabolismoRESUMO
Vibrio parahaemolyticus uses bacterial secretion systems and integrative and conjugative elements (ICEs) to induce various diseases and to adapt to harsh environments, respectively. Information pertaining to the identity of secreted proteins and functional characterization of ICEs has been previously reported, but the relationship between these elements remains unclear. Herein we investigated secreted proteins of V. parahaemolyticus strains JHY20 and JHY20â³ICE using two-dimensional gel electrophoresis and LC-MS/MS, which led to the identification of an ICE-associated secreted protein - dihydrolipoamide dehydrogenase (DLDH). Considering the data related to its physical and biochemical characterization, we predicted that DLDH is a novel immunogenic protein and associated with virulence in JHY20. Our findings indicate a potential relationship between ICE-associated transport and secreted proteins and shed light on the function of such transport mechanisms. We believe that our data should enhance our understanding of mobile genetic elements.
RESUMO
Serratia marcescens NJ01, a Gram-negative bacterium, can infect tomato leaves and cause chlorosis and wilting. The present study evaluated the quorum sensing (QS) and biofilm inhibitory effects of seven carboline compounds against S. marcescens NJ01 at 20 µg ml-1, and subsequently focused the study on norharmane as this had the best inhibitory activity. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis confirmed the down-regulation of QS and biofilm related genes bsmA, bsmB, fimA, fimC, flhD, pigA, pigC and shlA on exposure to norharmane. Fourier-Transform Infrared Spectroscopy (FT-IR) analysis showed a reduction in the major components of the exopolysaccharide (EPS) matrix such as nucleic acids, proteins and fatty acids, which are involved in forming the tertiary structure of biofilms. Norharmane exposure also enhanced the susceptibility of the biofilm to ofloxacin. Hence, norharmane has the potential for use as an antibiotic adjuvant to enhance the efficacy of conventional antibiotics to reduce pathogenic bacterial infections.
Assuntos
Percepção de Quorum , Serratia marcescens , Antibacterianos/farmacologia , Biofilmes , Carbolinas/farmacologia , Serratia marcescens/genética , Espectroscopia de Infravermelho com Transformada de Fourier , Fatores de Virulência/genéticaRESUMO
Phomopsis liquidambari is a classical endophytic fungus with great application potential in ecology and agriculture; however, studies on its exopolysaccharides are lacking. Here, we aimed to evaluate the structure and bioactivity of PLN-1, an exopolysaccharide derived from the P. liquidambari NJUSTb1 strain. The structure was elucidated by chromatography/spectral methods and hydrolyzation. Immunomodulation, moisture absorption, and retention properties were investigated after sulfation and carboxymethylation modification. Results showed that PLN-1 contained a linear repeating unit of â[4)-α-d-Glcp-(1â6)-α-d-Glcp-(1â4)-α-d-Glcp-(1â4)-α-d-Glcp-(1â]n, with a molecular weight of 343 kDa. The degrees of substitution of sulfated polysaccharide (S-PLN-1) and carboxymethylated polysaccharide (C-PLN-1) were 1.228 and 0.903, respectively. S-PLN-1 showed stronger moisture absorption and retention properties than PLN (crude EPS), C-PLN1, and PLN-1. Furthermore, PLN, S-PLN-1, and C-PLN-1 stimulated the proliferation of RAW 264.7 cells with no cytotoxicity. The elucidation of PLN-1 in this study paves the way for future applications.
Assuntos
Polissacarídeos Fúngicos/química , Polissacarídeos Fúngicos/farmacologia , Fatores Imunológicos/química , Fatores Imunológicos/farmacologia , Phomopsis/química , Absorção Fisico-Química , Animais , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Citocinas/metabolismo , Polissacarídeos Fúngicos/isolamento & purificação , Galactose , Glucose , Fatores Imunológicos/isolamento & purificação , Macrófagos/imunologia , Manose , Camundongos , Peso Molecular , Células RAW 264.7 , Transdução de Sinais/efeitos dos fármacosRESUMO
Phomopsis liquidambari S47 is an endophytic fungus isolated from the leaves of Punica granatum. Here, we are the first to report a quorum sensing (QS) inhibitor 1-(4-amino-2-hydroxyphenyl)ethanone (AHE) isolated and identified from the metabolites of P. liquidambari S47. Exposure to AHE at sub-MIC concentrations notably suppressed the secretion of acyl-homoserine lactones and virulence factors in Pseudomonas aeruginosa PAO1. To investigate the metabolic variations of P. aeruginosa PAO1 exposed to AHE, magnetic resonance imaging-based metabolomic analysis was performed. AHE treatment created a disturbance in the QS system by suppressing the expressions of QS-related genes. The disturbed QS system resulted in the inhibited activity of antioxidant enzymes and thus enhanced oxidative stress. The vegetable infection assay showed that the virulence of P. aeroginosa PAO1 was attenuated which could be due to the impacts to the amino acid and nucleotide metabolism by enhanced oxidative stress. These findings suggest that AHE has a potential to become an antivirulence "agent" to tackle P. aeruginosa infection. KEY POINTS: ⢠AHE treatment inhibited AHL secretion and virulence factors production. ⢠AHE treatment aggravated oxidative stress and disturbed metabolism. ⢠AHE suppressed QS-related gene expressions and reduced virulence of P. aeruginosa.
Assuntos
Antibiose , Phomopsis , Pseudomonas aeruginosa , Percepção de Quorum , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/farmacologia , Biofilmes , Virulência , Fatores de Virulência/genéticaRESUMO
The impact of 1-(4-amino-2-hydroxyphenyl)ethanone (AHPE) from the metabolites of endophytic fungus Phomopsis liquidambari on quorum sensing (QS) of Agrobacterium tumefaciens was evaluated for the first time in this study. Exposure to AHPE at concentrations ranging from 12.5 to 50 µg/mL, the ß-galactosidase activity, acyl-homoserine lactone level, swimming motility, chemotaxis, and flagella formation were significantly inhibited. qRT-PCR quantification combined with the docking analysis demonstrated that AHPE affected the QS system of A. tumefaciens by repressing the transcriptional levels of traI and traR rather than signal mimicry. 1H NMR-based metabolic analysis indicated that the metabolism of A. tumefaciens was notably disturbed with AHPE treatment. AHPE treatment also resulted in the enhanced oxidative stress in A. tumefaciens. The enhanced oxidative stress lead to the disorder of energy supply, protein synthesis, and nucleotide metabolism, and ultimately attenuated the pathogenicity of A. tumefaciens. Our study indicated that AHPE can serve as a potential pesticide to defend against A. tumefaciens.
RESUMO
Prosaikogenin D, a rare secondary saponin in Radix Bupleuri, has much higher in vivo bioactivities than its original glycoside saikosaponin B2. Its preparation methods, such as conventional acid hydrolysis and column chromatograph, are unfriendly to environment with serious pollution and undesired products. The aim of this study was to establish an efficient and clean approach for convenient preparation of this rare steroid saponin based on the enzymatic hydrolysis. Cellulase was selected from four commercial enzymes due to its higher hydrolysis performance. Then the hydrolysis conditions were optimized by response surface methodology after preliminary investigation on affecting factors by single-factor experiments. The reaction system was constructed by 100⯵g/mL of saikosaponin B2 and 8.00â¯mg/mL of cellulase, which was incubated in HAc-NaAc buffer (pH 4.7) at 60⯰C for 33â¯h. Consequently, a high conversion ratio of the substrate has been achieved at 95.04 %. The newly developed strategy is an efficient and clean approach for the preparation of prosaikogenin D and it is a promising technology in industrial application.