Quorum Sensing Signal Molecules Research Articles

A new strategy for greenhouse gas emission reduction in the anaerobic/anoxic/oxic biological treatment process using exogenous N-acyl-homoserine lactones, a quorum-sensing signaling molecules

ABSTRACT In this study, the impact of exogenous N-acyl-homoserine lactones (AHLs) on greenhouse gas (GHG) emissions in anaerobic/anoxic/oxic (A/A/O) systems was analyzed by manipulating the type and dosage of AHLs. The mechanism behind AHLs’ effects on GHG emissions was explored through changes in microbial community structure. Findings revealed that N-octanoyl-homoserine lactone (C8-HSL) and high-dose N-dodecanoyl-homoserine lactone (C12-HSL) increased GHG emissions, while low-dose C12-HSL decreased them. Moreover, C8-HSL and high-dose C12-HSL promoted methane (CH4) and nitrous oxide (N2O) production by affecting sludge particle size. Bacterial community analysis highlighted Acinetobacter and Flavobacterium's roles in N2O emissions and acetate methanogens in methane synthesis. Metabolic pathway analysis showed that the acetic acid (CH3COOH) methanogenic pathway was the main methanogenic pathway; C8-HSL and C12-HSL influenced methane emission by affecting the methanogenic pathway and N2O emission by changing nitrous oxide reductase (Nos) abundance. This research underscores AHL-based quorum sensing's potential in mitigating GHG emissions during activated sludge wastewater treatment, offering insights into their application and impact on key microbial activities. Limitations include the absence of methane emission reduction by signaling molecules and the need for further investigation into their effects on sludge accumulation.

Imipenem exposure influence the expression of quorum sensing receptor sdiA in Escherichia coli

Abstract The increasing trend of carbapenem resistance amongst Escherichia coli poses a major public health crisis and requires active surveillance of resistance mechanisms to control the threat. Quorum Sensing system plays a role in bacterial resistance to antibiotics. Quorum Sensing is a cell-cell communication system where bacteria alter their gene expression in response to specific stimuli. Here, in this study we investigated the transcriptional response of quorum sensing receptor, sdiA in E.coli under sub-inhibitory concentration of carbapenem in presence of quorum sensing signal molecules. Two E.coli isolates harbouring blaNDM were subjected to treatment with 10% SDS for 20 consecutive days of which blaNDM encoding plasmid was successfully eliminated from one isolate. Both the wild type and the cured mutant were then allowed to grow under eight different inducing conditions and the transcriptional response of sdiA gene was studied by quantitative real time PCR method. We found different response levels of sdiA in wild type and cured mutant under exogenous AHL and imipenem and when co-cultured with P.aeruginosa under imipenem stress. This study highlighted that sub-inhibitory concentration of imipenem in combination with AHL is acting as signal to SdiA, a quorum sensing receptor in E.coli.

Effects of different quorum sensing signal molecules on alleviation of ammonia inhibition during biomethanation

Anaerobic digestion (AD) is a promising technology for achieving both organic wastes treatment and energy recovery. However, challenges such as ammonia inhibition still remain. Quorum sensing (QS) system is relevant with the regulation of microbial community behaviors by releasing and sensing signal molecules, which could improve methane production during AD process. Therefore, the current study explored the effects of different quorum sensing signal molecules on alleviation of ammonia inhibition. The results showed that both secretion of N-butyryl-DL-homoserine lactone (C4-HSL) and N-(β-ketocaproyl)-DL-homoserine lactone (3OC6-HSL) could be inhibited by high ammonia stress while stimulation of N-hexanoyl-L-homoserine lactone (C6-HSL) and N-octanoyl-DL-homoserine lactone (C8-HSL) secretion might be triggered by ammonia toxicity. Moreover, the alleviation of ammonia inhibition could be achieved by both introducing 3OC6-HSL (0.5 μM) and combination of 3OC6-HSL (0.1 μM) and biochar (4 g/L). Exogenous 3OC6-HSL could regulate microbial social behaviors and enhance the secretion of extracellular polymeric substances (EPS) to promote anaerobic digestion. In addition, the mitigation of ammonia inhibition through exogenous 3OC6-HSL and biochar were confirmed by microbial community changes (Methanobacterium, Propionicicella and Petrimonas). Critical enzymes involved in both acidification and methanogenic steps were enhanced after adding the combination of 3OC6-HSL and biochar. The combination of low levels of 3OC6-HSL and biochar could promote both direct interspecies electron transfer (DIET) process and communication between different anaerobic microorganisms to mitigate ammonia inhibition. The current study will provide primary insights for conquering ammonia inhibition during biomethanation.

Efficient nitrogen removal and stable operation of a dynamic membrane bioreactor (DMBR) for landfill leachate treatment

This study proposed a novel anoxic/4-stage aerobic/self-forming dynamic membrane bioreactor (DMBR) process for treating landfill leachate with low carbon-to-nitrogen ratio (COD/N). When the influent COD and NH4+-N were 3689 ± 425 and 1042 ± 96 mg/L, high removal efficiencies of 87 % for COD and 79 % for TN can be achieved. The mature dynamic membrane (DM) was rapidly established within 1 h, allowing one operational cycle to last over 20 days with the permeate turbidity below 5 NTU. The primary cause of biofouling was identified as the accumulation of small particles, proteins, polysaccharides, and inorganic components such as Ca2+ and Mg2+ in the DM layer. In addition, significant up-regulation of genes associated with quorum sensing signaling molecules (such as pac, trpE, dgcB), extracellular polymeric substances secretion (exoP, exoQ, algA), and biofilm formation (lasA) were observed in the DM layer, while some genus, like Thauera, Anaerolinea and Caldilinea were highly enriched in DM layer. These findings indicate that DMBR offers cost-effective and technically advantageous operations, making it a promising candidate for the treatment of high-strength landfill leachate.

Global transcriptional response of Pseudomonas aeruginosa to UVA radiation.

Ultraviolet A (UVA) radiation is the major fraction of UV radiation reaching the Earth's surface. Its harmful effects on microorganisms, due mainly to oxidative damage, have been exploited for development of natural solar and commercial UVA-based disinfection methods. In this work, the global transcriptional response of Pseudomonas aeruginosa exposed to ultraviolet A (UVA) radiation was analyzed. To conduct this study, we analyzed the whole transcriptome of the PAO1 strain grown to logarithmic phase under sublethal doses of UVA or in the dark. We found that a total of 298 genes responded to UVA with a change of at least two-fold (5.36% of the total P. aeruginosa genome), and showed equal amount of induced and repressed genes. An important fraction of the induced genes were involved in the response to DNA damage and included induction of SOS, prophage and pyocins genes. The results presented in this study suggest that one of the main UVA targets are proteins carrying [Fe-S] clusters since several genes involved in the processes of synthesis, trafficking and assembly of these structures were upregulated. The management of intracellular iron levels also seems to be a robust response to this stress factor. The strong induction of genes involved in denitrification suggest that this pathway and/or reactive nitrogen species such as nitric oxide could have a role in the response to this radiation. Regarding the down-regulated genes, we found many involved in the biosynthesis of PQS, a quorum-sensing signal molecule with a possible role as endogenous photosensitizer.

Bacterial Acyl Homoserine Lactones Triggered Non‐Native Substrate Hydroxylation Catalyzed by Directed‐Evolution‐Derived Cytochrome P450BM3 Mutants

AbstractWe report the directed evolution of cytochrome P450BM3 to efficiently utilize the bacterial quorum sensing signalling molecule N‐decanoyl homoserine lactone (C10‐HSL) as an effective decoy molecule. This represents the first important step in our endeavor to develop of a self‐sufficient decoy‐molecule system in whole‐cells that only necessitates the addition of culture medium and substrate to realize the hydroxylation of non‐native substrates. Following five rounds of directed evolution, mutant P450BM3, in the presence of C10‐HSL, catalyzed the hydroxylation of benzene at a rate of 475 min−1, the highest turnover rate recorded for any P450 enzyme, and achieving a 46% yield in a whole‐cell reaction system. High‐resolution X‐ray crystal structure analysis of a series of mutants narrates the directed evolution process, revealing how C10‐HSL is fixed in the binding pocket to permit binding of non‐native substrates. Finally, introduction of the C10‐HSL synthase gene ExpI into Escherichia coli, enabled the in situ production of C10‐HSL, realizing, for the first time, the hydroxylation of non‐native substrates without the need for the laborious synthesis and addition of decoy molecules.

In Situ Enrichment of Anammox Bacteria from Pig Farm Anoxic Sludge Through Co-Cultivation with a Quorum-Sensing Functional Strain Pseudomonas aeruginosa

Anaerobic ammonium oxidation (anammox), as an efficient and low-carbon method for nitrogen removal from wastewater, faces the challenge of slow enrichment of functional bacteria. In this study, the enrichment of anammox bacteria Candidatus Brocadia was successfully accelerated by co-culturing with the quorum-sensing strain Pseudomonas aeruginosa and anoxic sludge from a pig farm. Experimental results showed that the R2, which had Pseudomonas aeruginosa added, exhibited chemical reaction ratios RS (NO2−-N consumption/NH4+-N consumption) and RP (NO3−-N production/NH4+-N consumption) closer to the theoretical values of the anammox reaction since Phase Ⅱ. Bacterial community analysis indicated that the abundance of Candidatus Brocadia in R2 reached 1.63% in cycle 20, significantly higher than the 0.45% in R1. More quorum-sensing signaling molecules, primarily C6-HSL, were detected in R2. C6-HSL was positively correlated with processes such as the secretion of anammox extracellular polymers (EPS) and the regulation of nitric oxide reductase (Nir), which may explain the reason behind the accelerated increase in the abundance of Candidatus Brocadia through co-culturing. Moreover, the metabolism of the dominant genus Paracoccus within the two groups of reactors also showed positive regulation by C6-HSL, with its abundance trend similar to that of Candidatus Brocadia, jointly completing the nitrogen removal process in the reactors. However, it is still unknown which genera secrete large amounts of C6-HSL after inoculation with Pseudomonas aeruginosa. This research provides a novel and low-cost method for the enrichment of anammox bacteria.

Quorum Sensing Molecule Autoinducer-2 Promotes Macrophage Classical Polarization and Exacerbates Periodontal Inflammation Via Nf-Κb Signalling.

The role of quorum sensing signaling in the immunoinflammatory response during the development of periodontitis is not yet known. This study aimed to explore the effect of Autoinducer-2, a quorum sensing signaling molecule, on macrophage phenotypic remodeling in the immune microenvironment of periodontitis, to further elucidate its mechanism and to discover inhibitors against periodontitis. Bioluminescence experiments and periodontitis model were used to demonstrate the association between periodontitis progression with AI-2. Next, AI-2 challenged macrophage was introduced to transcriptomic sequence and the immune profile was characterized in combination with flow cytometry, qPCR, and immunofluorescence. Activation of NF-κB signalling by AI-2 was confirmed by fluorescence co-localization and immunoblotting. Finally, morphological methods such as Micro-CT and HE, TRAP staining and immunological methods such as immunohistochemistry/fluorescence staining were used to assess the mechanisms by which AI-2 regulates periodontitis progression. AI-2 level was positively correlated with the progression of periodontitis stages and was significantly higher in periodontitis stage III and IV patients. AI-2 promotes macrophage classical polarization and facilitates the secretion of inflammatory factors in vitro, which is dependent on the activation of the NF-κB signaling pathway. AI-2 promotes alveolar bone resorption, but D-ribose acts as a quorum sensing inhibitor to alleviate macrophage classical polarization and attenuates alveolar bone resorption and inflammatory responses in periodontitis mice. Our study demonstrates that AI-2 promoted classical polarization of macrophage and exacerbated periodontal inflammation which could be reversed by D-ribose.

Metabolic complexities and heterogeneity in quorum sensing signaling molecules in bacteria isolated from black band disease in a Caribbean coral

Coral diseases contribute to the worldwide loss of coral reefs, with the Black Band Disease (BBD) being a prominent example. BBD is an infectious condition with lesions with a pigmented mat composed of cyanobacteria, sulphate-reducing, sulphide-oxidizing, and heterotrophic bacteria. We compared the heterotrophic bacterial communities of healthy and BBD-affected colonies of the Caribbean coral Orbicella faveolata using culture-dependent and -independent techniques. Twenty and 23 bacterial isolates were identified from healthy and diseased tissues, respectively, which differed in their capacities to metabolize carbohydrates and citrate, either anaerobically or aerobically. They also differed in their quorum-sensing (QS) activity, as QS signaling molecules were found exclusively, and QS-inhibition was found primarily, in isolates from diseased tissues. Screening of bacterial diversity by 16SrDNA metabarcoding showed that members of the bacterial genera Muricauda and Maritimimonas were dominant in healthy tissues whereas members of the cyanobacterial genus Roseofilum were dominant in diseased tissues. These results suggest that bacterial dysbiosis can be linked with altered bacterial communication, likely leading to diachrony and imbalance that may participate in the progression of BBD. Investigating physiological traits and QS-based communication offers insights into the onset and progression of coral infections, paving the way for novel strategies to mitigate their impact.

Fatty acid synthesis promoted by PA1895-1897 operon delays quorum sensing activation in Pseudomonas aeruginosa

PA1895-1897 is a quorum sensing (QS) operon regulated by the anti-activator LuxR homologue QscR in Pseudomonas aeruginosa. We aimed to investigate its impact on bacterial metabolism, and whether it contributes to the delayed QS activation. We performed liquid chromatograph-mass spectrometer-based metabolomics using wildtype PAO1, PA1895-1897-knockout mutant, and mutant with pJN105.PA1895-1897 overexpression vector. The impact of metabolites on QS signaling molecule (3OC12-HSL and C4-HSL) concentrations, pyocyanin production, and QS gene (lasR, lasI, rhlR, and rhlI) expression was examined. Metabolomics analysis found that fatty acid biosynthesis had the highest fold enrichment among all metabolic pathways in PA1895-1897-overexpressed mutants. Among these enriched fatty acids, palmitoleic acid and acetic acid were the predominantly abundant ones that significantly affected by PA1895-1897 operon. When different doses of exogenous palmitoleic acid or acetic acid were added to the cultures of PA1895-1897 knockout mutants, their levels of 3OC12-HSL, C4-HSL, and pyocyanin were decreased in a dose-dependent manner. High doses of palmitoleic acid and acetic acid suppressed the mRNA expression of lasR, rhlR, and rhlI. Inhibition of fatty acid biosynthesis increased the production of 3OC12-HSL, C4-HSL, and pyocyanin in PA1895-1897-overexpressed cultures. Our data suggest that fatty acid synthesis is promoted by PA1895-1897 operon, and contributes the delayed expression of QS phenotypes, furthering the understanding about the regulation of bacterial QS activation.

Effect of the quorum sensing signal molecule auto-inducer-2 on fermentation performance of Lactobacillus delbrueckii subsp. Bulgaricus DPUL-F36

Fermented milk is a typically product of co-fermentation by a variety of lactic acid bacteria (LAB). Quorum sensing (QS), as an intercellular communication mechanism of bacteria, may serve as a key factor influencing the quality of fermented milk. In this study, the fermentation performance of Lactobacillus delbrueckii subsp. bulgaricus DPUL-F36 were evaluated after supplementation with auto-inducer-2 (AI-2). The protein-protein interaction network (PPIN) was used to retrieve the interactions between QS genes and four major metabolic pathway genes. The expression of luxS gene in DPUL-F36 may affect amino acid metabolism, carbohydrate metabolism and cell wall formation. The key differential metabolites were identified by metabolomics analysis. A variety of amino acids including L-Glutamic, L-Tryptophan, L-Threonine, L-Histidine and L-Glutamine were found to be up-regulated in response to the addition of AI-2. Malic acid and succinate gradually accumulate in the tricarboxylic acid cycle. The consumption of citric acid increased with the fermentation process. In terms of rheological and texture characteristics of fermented milk, addition of AI-2 could effectively improve its shear stress resistance, firmness and index of viscosity. Our research provided a theoretical basis for improving the fermentation performance of DPUL-F36 strains, and support for the screening of excellent composite starter culture.

The bacterial quorum sensing signal 2'-aminoacetophenone rewires immune cell bioenergetics through the Ppargc1a/Esrra axis to mediate tolerance to infection.

How bacterial pathogens exploit host metabolism to promote immune tolerance and persist in infected hosts remains elusive. To achieve this, we show that Pseudomonas aeruginosa (PA), a recalcitrant pathogen, utilizes the quorum sensing (QS) signal 2'-aminoacetophenone (2-AA). Here, we unveil how 2-AA-driven immune tolerization causes distinct metabolic perturbations in murine macrophages' mitochondrial respiration and bioenergetics. We present evidence indicating that these effects stem from decreased pyruvate transport into mitochondria. This reduction is attributed to decreased expression of the mitochondrial pyruvate carrier (Mpc1), which is mediated by diminished expression and nuclear presence of its transcriptional regulator, estrogen-related nuclear receptor alpha (Esrra). Consequently, Esrra exhibits weakened binding to the Mpc1 promoter. This outcome arises from the impaired interaction between Esrra and the peroxisome proliferator-activated receptor gamma coactivator 1-alpha (Ppargc1a). Ultimately, this cascade results in diminished pyruvate influx into mitochondria and, consequently reduced ATP production in tolerized murine and human macrophages. Exogenously added ATP in infected macrophages restores the transcript levels of Mpc1 and Esrra and enhances cytokine production and intracellular bacterial clearance. Consistent with the in vitro findings, murine infection studies corroborate the 2-AA-mediated long-lasting decrease in ATP and acetyl-CoA and its association with PA persistence, further supporting this QS signaling molecule as the culprit of the host bioenergetic alterations and PA persistence. These findings unveil 2-AA as a modulator of cellular immunometabolism and reveal an unprecedented mechanism of host tolerance to infection involving the Ppargc1a/Esrra axis in its influence on Mpc1/OXPHOS-dependent energy production and PA clearance. These paradigmatic findings pave the way for developing treatments to bolster host resilience to pathogen-induced damage. Given that QS is a common characteristic of prokaryotes, it is likely that 2-AA-like molecules with similar functions may be present in other pathogens.

Complete genome sequence provides information on quorum sensing related spoilage and virulence of Aeromonas salmonicida GMT3 isolated from spoiled sturgeon

Foodborne bacteria can pose a threat to the public health due to their spoilage and virulence potential, which can be regulated by quorum sensing (QS) system. In the study, we isolated a spoilage bacteria strain Aeromonas salmonicida GMT3 from refrigerated sturgeon. The complete genome of A. salmonicida GMT3 was sequenced, and the QS related genes were assigned. QS signal molecules N-acyl-homoserine lactones (AHLs) and AI-2 were detected. Genes regulating the spoilage-related metabolic pathways, including protease and lipase secretion, amines metabolism, sulfur metabolism, motility and biofilm formation were analyzed. Furthermore, genes encoding for several virulence factors, e.g. hemolysin, aerolysin, type II secretion system (T2SS), type VI secretion system (T6SS), antibiotic and multidrug resistance were also identified. In addition, the spoilage and virulence phenotypes associated with QS including protease, swimming and swarming activity, biofilm and hemolytic activity were detected. This study provided new insights into spoilage and virulence mechanisms correlated with QS of A. salmonicida GMT3, which might promote development of new approaches for spoilage and virulence control based on QS target.

The diadenosine tetraphosphate hydrolase ApaH contributes to Pseudomonas aeruginosa pathogenicity.

The opportunistic bacterial pathogen Pseudomonas aeruginosa causes a wide range of infections that are difficult to treat, largely because of the spread of antibiotic-resistant isolates. Antivirulence therapy, í.e. the use of drugs that inhibit the expression or activity of virulence factors, is currently considered an attractive strategy to reduce P. aeruginosa pathogenicity and complement antibiotic treatments. Because of the multifactorial nature of P. aeruginosa virulence and the broad arsenal of virulence factors this bacterium can produce, the regulatory networks that control the expression of multiple virulence traits have been extensively explored as potential targets for antivirulence drug development. The intracellular signaling molecule diadenosine tetraphosphate (Ap4A) has been reported to control stress resistance and virulence-related traits in some bacteria, but its role has not been investigated in P. aeruginosa so far. To fill this gap, we generated a mutant of the reference strain P. aeruginosa PAO1 that lacks the Ap4A-hydrolysing enzyme ApaH and, consequently, accumulates high intracellular levels of Ap4A. Phenotypic and transcriptomic analyses revealed that the lack of ApaH causes a drastic reduction in the expression of several virulence factors, including extracellular proteases, elastases, siderophores, and quorum sensing signal molecules. Accordingly, infection assays in plant and animal models demonstrated that ApaH-deficient cells are significantly impaired in infectivity and persistence in different hosts, including mice. Finally, deletion of apaH in P. aeruginosa clinical isolates demonstrated that the positive effect of ApaH on the production of virulence-related traits and on infectivity is conserved in P. aeruginosa. This study provides the first evidence that the Ap4A-hydrolysing enzyme ApaH is important for P. aeruginosa virulence, highlighting this protein as a novel potential target for antivirulence therapies against P. aeruginosa.

Targeting N-acyl-homoserine lactones (AHLs) on the quorum quenching AiiA protein to eradicate biofilm formation in Vibrio parahaemolyticus

ABSTRACT AiiA can be targeted to quench N-acyl-homoserine lactones (AHLs) quorum sensing signaling molecules. In this paper, we extracted and purified AiiA protein by constructing recombinant engineering bacteria to analyze the effect of AiiA on the formation of Vibrio parahaemolyticus biofilm. The results suggest that AiiA has a particular potential in inhibiting the formation of biofilms and extracellular polymers. The ability of AiiA to inhibit biofilm increased and then decreased, with the lowest biofilm decomposition concentration of 8 μg/mL and the optimal inhibitor concentration of 85 μg/mL (86.9% biofilm inhibition). LC-MS results showed that when the concentration of C4-HSL in Vibrio parahaemolyticus ATCC33847 biofilm exceeded 3.70 μg/L, the content of quorum sensing signaling molecules reached a threshold and began to activate LuxR protein to regulate the expression of related genes. These results provide relevant experimental guidance for the clinical treatment of biofilm infections and the development of novel QS inhibitors.

Collimonas rhizosphaerae sp. nov., a novel species isolated from the beech rhizosphere.

Bacterial strain H4R21T was isolated from beech rhizosphere soil sampled in the forest experimental site of Montiers (Meuse, France). It effectively weathers minerals, hydrolyses chitin and produces quorum sensing signal molecules. The strain is aerobic and Gram-stain-negative. Phylogenetic analysis based on its 16S rRNA gene sequence indicated that strain H4R21T belongs to the genus Collimonas with high sequence similarity to C. arenae Ter10T (99.38 %), C. fungivorans Ter6T(98.97 %), C. pratensis Ter91T (98.76 %), C. humicola RLT1W51T (98.46 %) and C. silvisoli RXD178 T (98.46 %), but less than 98 % similarity to other strains of the genus Collimonas. The predominant quinone in H4R21T is ubiquinone-8 (Q8). The major polar lipids are diphosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol and lipid. The major fatty acids identified were C12 : 0, C12:0 3-OH, C16 : 0 and C17:0 cyclo. The digital DNA G+C content of the genomic DNA was 59.5 mol%. Furthermore, the strain could be clearly distinguished from its closely related type strains by a combination of phylogenomic and in silico DNA-DNA hybridization results, and phenotypic characteristics. Therefore, strain H4R21T represents a novel species within the genus Collimonas, for which the name Collimonas rhizosphaerae sp. nov. is proposed, with strain H4R21T (=CFBP 9203T=DSM 117599T) as the type strain.

Dopamine, an exogenous quorum sensing signaling molecule or a modulating factor in Pseudomonas aeruginosa?

Pseudomonas aeruginosa is recognized globally as an opportunistic pathogen of considerable concern due to its high virulence and pathogenicity, especially in immunocompromised individuals. While research has identified several endogenous quorum sensing (QS) signaling molecules that enhance the virulence and pathogenicity of P. aeruginosa, investigations on exogenous QS signaling molecules or modulating factors remain limited. This study found that dopamine serves as an exogenous QS signaling molecule or modulating factor of P. aeruginosa PAO1, enhancing the production of virulence factors and biofilms. Compared to the control group, treatment with 40 μM dopamine resulted in a 33.1 % increase in biofilm formation, 68.1 % increase in swimming mobility, 63.1 % increase in swarming mobility, 147.2 % increase in the signaling molecule 3-oxo-C12-HSL, and 50.5 %, 28.5 %, 27.0 %, and 33.2 % increases in the virulence factors alginate, rhamnolipids, protease, and pyocyanin, respectively. This study further explored the mechanism of dopamine regulating the biofilm formation and virulence of P. aeruginosa PAO1 through transcriptome and metabolome. Transcriptomic analysis showed that dopamine promoted the expression of virulence genes psl, alg, lasA, rhlABC, rml, and phz in P. aeruginosa PAO1. Metabolomic analysis revealed changes in the concentrations of tryptophan, pyruvate, ethanolamine, glycine, 3-hydroxybutyric acid, and alizarin. Furthermore, KEGG enrichment analysis of altered genes and metabolites indicated that dopamine enhanced phenylalanine, tyrosine, and tryptophan in P. aeruginosa PAO1. The results of this study will contribute to the development of novel exogenous QS signaling molecules or modulating factors and advance our understanding of the interactions between P. aeruginosa and the host environment.

The Bacterial Quorum-Sensing Signal 2-Aminoacetophenone Rewires Immune Cell Bioenergetics through the PGC-1α/ERRα Axis to Mediate Tolerance to Infection.

How bacterial pathogens exploit host metabolism to promote immune tolerance and persist in infected hosts remains elusive. To achieve this, we show that Pseudomonas aeruginosa (PA), a recalcitrant pathogen, utilizes the quorum sensing (QS) signal 2-aminoacetophenone (2-AA). Here, we unveil how 2-AA-driven immune tolerization causes distinct metabolic perturbations in macrophages mitochondrial respiration and bioenergetics. We present evidence indicating that these effects stem from decreased pyruvate transport into mitochondria. This reduction is attributed to decreased expression of the mitochondrial pyruvate carrier (MPC1), which is mediated by diminished expression and nuclear presence of its transcriptional regulator, estrogen-related nuclear receptor alpha (ERRα). Consequently, ERRα exhibits weakened binding to the MPC1 promoter. This outcome arises from the impaired interaction between ERRα and the peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α). Ultimately, this cascade results in diminished pyruvate influx into mitochondria and, consequently reduced ATP production in tolerized macrophages. Exogenously added ATP in infected macrophages restores the transcript levels of MPC1 and ERRα and enhances cytokine production and intracellular bacterial clearance. Consistent with the in vitro findings, murine infection studies corroborate the 2-AA-mediated long-lasting decrease in ATP and acetyl-CoA and its association with PA persistence, further supporting this QS signaling molecule as the culprit of the host bioenergetic alterations and PA persistence. These findings unveil 2-AA as a modulator of cellular immunometabolism and reveal an unprecedented mechanism of host tolerance to infection involving the PGC-1α/ERRα axis in its influence on MPC1/OXPHOS-dependent energy production and PA clearance. These paradigmatic findings pave the way for developing treatments to bolster host resilience to pathogen-induced damage. Given that QS is a common characteristic of prokaryotes, it is likely that 2-AA-like molecules with similar functions may be present in other pathogens.

Effects of chlorogenic acid-grafted-chitosan on biofilms, oxidative stress, quorum sensing and c-di-GMP in Pseudomonas fluorescens

This study determined the inhibitory mechanism as well as anti-biofilm activity of chlorogenic acid-grafted-chitosan (CS-g-CA) against Pseudomonas fluorescens (P. fluorescens) in terms of biofilm content, oxidative stress, quorum sensing and cyclic diguanosine monophosphate (c-di-GMP) concentration, and detected the changes in the expression levels of related genes by quantitative real-time PCR (qRT-PCR). Results indicated that treatment with sub-concentrations of CS-g-CA for P. fluorescens led to reduce the biofilm size of large colonies, decrease the content of biofilm and extracellular polymers, weaken the motility and adhesion of P. fluorescens. Moreover, CS-g-CA resulted in higher ROS levels, diminished catalase activity (CAT), and increased superoxide dismutase (SOD) in P. fluorescens. CS-g-CA reduced the production of quorum-sensing signaling molecules (AHLs) and the concentration of c-di-GMP in bacteria. Genes for flagellar synthesis (flgA), the resistance to stress (rpoS and hfq), and pde (phosphodiesterases that degrade c-di-GMP) were significantly down-regulated as determined by RT-PCR. Overall, CS-g-CA leads to the accumulation of ROS in bacteria via P. fluorescens environmental resistance genes and decreases the activity of enzymes in the bacterial antioxidant system, and interferes with the production and reception of quorum-sensing signaling molecules and the synthesis of c-di-GMP in P. fluorescens, which regulates the generation of biofilms.

MzmL, a novel marine derived N-acyl homoserine lactonase from Mesoflavibacter zeaxanthinifaciens that attenuates Pectobacterium carotovorum subsp. carotovorum virulence.

Quorum sensing (QS) is a conserved cell-cell communication mechanism widely distributed in bacteria, and is oftentimes tightly correlated with pathogen virulence. Quorum quenching enzymes, which interfere with QS through degrading the QS signaling molecules, could attenuate virulence instead of killing the pathogens, and thus are less likely to induce drug resistance. Many Gram-negative bacteria produce N-acyl homoserine lactones (AHLs) for interspecies communication. In this study, we isolated and identified a bacterial strain, Mesoflavibacter zeaxanthinifaciens XY-85, from an Onchidium sp. collected from the intertidal zone of Dapeng Reserve in Shenzhen, China, and found it had strong AHL degradative activity. Whole genome sequencing and blast analysis revealed that XY-85 harbors an AHL lactonase (designated MzmL), which is predicted to have an N-terminal signal peptide and share the "HXHXDH" motif with known AHL lactonases belonging to the Metallo-β-lactamase superfamily. Phylogenetic studies showed MzmL was closest to marine lactonase cluster members, MomL and Aii20J, instead of the AiiA type lactonases. Ultra performance liquid chromatography-mass spectrometry analysis confirmed that MzmL functions as an AHL lactonase catalyzing AHL degradation through lactone hydrolysis. MzmL could degrade both short- and long-chain AHLs with or without a substitution of oxo-group at the C-3 position, and retained full bioactivity under a wide range of temperatures (28-100°C) and pHs (4-11). Furthermore, MzmL significantly reduced Pectobacterium carotovorum subsp. carotovorum virulence factor production in vitro, such as biofilm formation and plant cell wall degrading enzyme production, and inhibited soft rot development on potato slices. These results demonstrated that MzmL may be a novel type of AHL lactonase with good environmental stability, and has great potential to be developed into a novel biological control agent for bacterial disease management.