Quorum Quenching Activity Research Articles

Benefits of fungal-to-bacterial quorum quenching as anti-biofouling strategy in membrane bioreactors for wastewater treatment and water reuse

This study addresses membrane biofouling in membrane bioreactors (MBRs) by exploring fungal-to-bacterial quorum quenching (QQ) strategies. While most research has been focused on bacterial-to-bacterial QQ tactics, this study identified fungal strain Vanrija sp. MS1, which is capable of degrading N-acyl-homoserine lactones (signaling molecules of Gram-negative bacteria). To determine the benefits of fungal over bacterial strains, after immobilization on fluidizing spherical beads in an MBR, MS1 significantly reduced the fouling rate by 1.8-fold compared to control MBR, decreased extracellular polymeric substance levels in the biofilm during MBR operation, and favorably changed microbial community and bacterial network, resulting in biofouling mitigation. It is noteworthy that, unlike Rhodococcus sp. BH4, MS1 enhanced QQ activity when switching from neutral to acidic conditions. These results suggest that MS1 has the potential for the effective treatment of acidic industrial wastewater sources such as semiconductor and secondary battery wastewater using MBRs.

Preservation and reactivation strategies for quorum quenching media to combat membrane biofouling

Microbial quorum quenching (QQ), which addresses biofouling resistant to physical and chemical methods, presents a compelling solution. Enhancements in sustainability are imperative to ensure its long-term stability. This study explores QQ media preservation for extended use. Three media types were tested over 150 days: fresh BH4 cells, preserved/reactivated BH4 cells, and BH4 cell extracts. Initial QQ activities ranged in the appropriate levels from 1.08 to 1.24 h−1. Preservation in 50 mM Tris-HCl at 4 °C showed that media with whole cells retained approximately 60 % activity after 60 days, unlike cell extract media, which lost all activity. However, the dehydration and reactivation of BH4 cell media fully recovered the initial QQ activity while maintaining it for 150 days. In MBR tests, reactivated QQ media performed as effectively as those prepared with freshly cultured BH4 cells, delaying membrane fouling by 2-fold. Revitalizing QQ media in combination with chlorine-based chemically enhanced backwashing (CEB) further delayed fouling by 2.5 times compared to CEB alone. Reactivated QQ media retained robust activity even after prolonged MBR use. This study showcases the effectiveness of dehydration and reactivation techniques for preserving QQ media, ensuring its reliable long-term storage and utilization. However, further enhancements are necessary to optimize cell viability.

Development of a novel membrane-based quorum-quenching microbial isolator for biofouling control: Process performance and microbial mechanism

Quorum quenching (QQ) can mitigate biofouling in membrane bioreactors (MBRs) by inhibiting cell-to-cell communication. However, it is difficult to maintain long-term QQ activity. Here, a novel microbial isolator composed of tubular microfiltration membranes was developed to separate QQ bacteria (Rhodococcus sp. BH4) from sludge. The time to reach a transmembrane pressure of 50 kPa was delayed by 69.55 % (p = 0.002, Student’s t test) in MBR with QQ microbial isolator (MBR-Q), compared to that in the control MBR (MBR-C) during stable operation. The concentration of proteins in the extracellular polymeric substances of sludge was reduced by 20.61 % in MBR-Q relative to MBR-C. The results of the bacterial community analyses indicated less enrichment of fouling-associated bacteria (e.g., Acinetobacter) but a higher abundance of QQ enzymes in MBR-Q than in MBR-C. This environmentally friendly technique can decrease the cleaning frequency and increase the membrane lifespan, thus improving the sustainability of MBR technology.

Mitigation of biofouling in membrane bioreactors by quorum-quenching bacteria during the treatment of metal-containing wastewater.

Quorum quenching (QQ) is an efficient way to mitigate membrane biofouling in a membrane bioreactor (MBR) during wastewater treatment. A QQ bacterium, Lysinibacillus sp. A4, was isolated and used to mitigate biofouling in an MBR during the treatment of wastewater containing metals. A QQ enzyme (named AilY) was cloned from A4 and identified as a metallo-β-lactamase-like lactonase. The QQ activity of A4 and that of Escherichia coli BL21 (DE3) overexpressing AilY could be promoted by Fe2+, Mn2+, and Zn2+ while remaining unaffected by other metals tested. The two bacteria effectively mitigated biofouling by reducing the transmembrane pressure from around 30 to 20kPa without negative influence on the COD, NH4+-N, or total phosphorus of the effluent. The relative abundance of Lysinibacillus sp. A4 increased greatly from 0.04 to 8.29% in the MBR with metal-containing wastewater, suggesting that Lysinibacillus sp. A4 could multiply quickly and adapt to this environment. Taken together, the findings suggested that A4 could tolerate metal to a certain degree, and this property could allow A4 to adapt well to metal-containing wastewater, making it a valuable strain for mitigating biofouling in MBR during the treatment of metal-containing wastewater.

Exploring novel quorum quenching strain: Enhanced disrupting autoinducer-2 bacterial communication to combat biofouling in membrane bioreactor for wastewater treatment

Prior investigations concerning quorum quenching (QQ), which hinders quorum sensing (QS) in microbial communication, concentrated predominantly on N-acylhomoserine lactones (AHLs), which are communicated merely by gram-negative bacteria. However, to combat biofouling more effectively, particularly biofilm-related issues, it is important to explore the inhibition of autoinducer-2 (AI-2), a universal signal (interspecies communication) employed by both gram-negative and gram-positive bacteria. This study aimed to isolate and identify novel strains capable of more effectively inhibiting AI-2 signaling for use in engineering fields. The results revealed that the newly isolated strain Pantoea sp. PL-1 demonstrated remarkable efficacy in attenuating AI-2 signals, leading to a substantial reduction in the AI-2 precursor (S)-4,5-dihydroxy-2,3-pentandione (DPD) in both its pellet (with a DPD removal efficiency of 90.8 % after 180 min, with a rate constant of k = 0.8 h−1) and supernatant form (66.9 %, k = 0.37 h−1). Interestingly, the findings indicate that strain DKY-1, a strain previously reported to exhibit AI-2 QQ activity, only reduced DPD levels in its supernatant form (extracellular activity), while both the pellet and the supernatant of strain PL-1 demonstrated AI-2 QQ activity, indicating both intracellular and extracellular QQ activity. Adding the PL-1 supernatant to co-cultures of two AI-2 QS strains, E. coli K12 and P. mirabilis, successfully reduced DPD by 40 % and 70 %, respectively, without impeding their growth. Additionally, when assessing the impact of PL-1 to minimize membrane biofouling in a membrane bioreactor, it demonstrated superior performance compared to the DKY-1 strain, with a 40 % improvement. The AI-2 QQ compound of PL-1 has been identified as highly hydrophilic and low-molecular-weight, and its exact properties have not yet been fully elucidated. Therefore, uncovering its identity may be an important goal for future research.

Potential of the quorum-quenching and plant-growth promoting halotolerant Bacillus toyonensis AA1EC1 as biocontrol agent.

The use of fertilizers and pesticides to control plant diseases is widespread in intensive farming causing adverse effects together with the development of antimicrobial resistance pathogens. As the virulence of many Gram-negative phytopathogens is controlled by N-acyl-homoserine lactones (AHLs), the enzymatic disruption of this type of quorum-sensing (QS) signal molecules, mechanism known as quorum quenching (QQ), has been proposed as a promising alternative antivirulence therapy. In this study, a novel strain of Bacillus toyonensis isolated from the halophyte plant Arthrocaulon sp. exhibited numerous traits associated with plant growth promotion (PGP) and degraded a broad range of AHLs. Three lactonases and an acylase enzymes were identified in the bacterial genome and verified invitro. The AHL-degrading activity of strain AA1EC1 significantly attenuated the virulence of relevant phytopathogens causing reduction of soft rot symptoms on potato and carrots. Invivo assays showed that strain AA1EC1 significantly increased plant length, stem width, root and aerial dry weights and total weight of tomato and protected plants against Pseudomonas syringae pv. tomato. To our knowledge, this is the first report to demonstrate PGP and QQ activities in the species B. toyonensis that make this strain as a promising phytostimulant and biocontrol agent.

Carbon metabolism characteristics of quorum quenching bacteria Rhodococcus sp. BH4 determine the bioaugmentation efficiency under different carbon source conditions

Carbon sources are critical factors influencing bacterial bioaugmentation, however, the underlying mechanisms, particularly the metabolic characteristics of bioaugmented bacteria remain poorly understood. The bioaugmented bacterium Rhodococcus sp. BH4 secretes the quorum quenching (QQ) enzyme QsdA to disrupt the quorum sensing (QS) in the activated sludge (AS) process, reducing AS yield in-situ. This study investigated the carbon metabolic characteristics of BH4 and explored the effects on bioaugmentation with different influent carbon sources. Because of the absence of glucose-specific phosphoenol phosphotransferase system (PTS), BH4 prefers sodium acetate to glucose. However, the lactones produced during extracellular glucose metabolism enhance BH4 qsdA expression. Moreover, BH4 possess carbon catabolite repression (CCR), acetate inhibits glucose utilization. BH4 microbeads were added to reactors with different carbon sources (R1: sodium acetate; R2: glucose; R3: a mixture of sodium acetate and glucose) for in-situ AS yield reduction. During operation, AS reduction efficiency decreased in the following order: R1 > R3 > R2. R2 and R3 microbeads exhibited similar QQ activity to R1, with less BH4 biomass at 5 d. 13C labeling and Michaelis-Menten equation showed that, due to differences in the competitiveness of carbon sources, R1 BH4 obtained the most carbon, whereas R2 BH4 obtained the least carbon. Moreover, acetate inhibited glucose utilization of R3 BH4. Transcriptome analysis showed that R1 BH4 qsdA expression was the lowest, R2 BH4 was the most serious form of programmed cell death, and the R3 BH4 PTS pathway was inhibited. At 10 d, R1 BH4 biomass and microbead QQ activity were higher than that in R3, and the R2 BH4 lost viability and QQ activity. This study provides new insights into bioaugmentation from the perspectives of carbon source competitiveness, carbon metabolism pathways, and CCR.

Isolation and application of Bacillus thuringiensis LZX01: Efficient membrane biofouling mitigation function and anti-toxicity potential

Significant progress has been made in mitigating membrane biofouling by microbial quorum quenching (QQ). More efficient and survivable QQ strains need to be discovered. A new strain named Bacillus thuringiensis LZX01 was isolated in this study using a low carbon source concentration “starving” method from a membrane bioreactor (MBR). LZX01 secreted intracellular lactonase to enable QQ behavior and was capable of degrading 90 % of C8-HSL (200 ng/mL) within 30 min, which effectively delayed biofouling by inhibiting the growth of bacteria associated with biofouling and improving the hydrophilicity of bound extracellular polymeric substances. As a result, the membrane biofouling rate of MBR adding LZX01 was four times slower than that of the control MBR. Importantly, LZX01 maintains its QQ activity even in environments contaminated with typical toxic pollutants. Therefore, with high efficiency, toxicity resistance, and easy culture, LZX01 holds great potential and significant promise for biofouling control applications.

Catechol-type siderophore producer Streptomyces spp. with quorum-quenching activity isolated from wild medicinal plants rhizosphere

Traditional medicinal plants represent a unique source for the isolation of Streptomycetes and antimicrobial compounds. Antimicrobial activity of Streptomyces isolates collected from the rhizosphere of different native medicinal plants in Iran, was investigated against Gram-positive and Gram-negative bacteria. Multi-omics analysis was performed to identify active compounds corresponding to the observed bioactivities. Since cell-to-cell communication mediated by N-acyl homoserine lactones (AHLs) is important for the virulence of Gram-negative pathogenic bacteria, quorum quenching activity of the isolates and their ability to degrade C6-HSL and C8-HSL were tested. Two highly related Streptomyces isolates derived from the Helichrysum rubicundum and Rumex acetosa rhizosphere, respectively, exhibited bioactivity against pathogenic indicator bacteria. Metabolite analysis of extracts obtained from liquid cultures of both isolates revealed the production of the catechol-peptide siderophores streptobactin and tribenarthin. Proteomic analysis confirmed the presence of proteins encoded by the streptobactin and lidamycin biosynthetic gene clusters. Quorum quenching activity of the strains and AHL-degrading enzyme production was confirmed using a GFP producing biosensor and LC-MS analysis. This study demonstrates catecholate siderophore production and QQ potential of two Streptomyces isolates from medicinal plants rhizosphere from Mishan Plain, Hamedan Province, Iran using metabolomics and proteomics analysis. Production of streptobactin by terrestrial Streptomyces sp. strains was confirmed for the first time.

Quorum quenching activity of endophytic Bacillus sp. EBS9 from Tecomella undulata and its biocontrol applications

This study investigates the quorum quenching (QQ) activity of an endophytic bacterium, Bacillus sp. EBS9, isolated from the native medicinal plant Tecomella undulata of Rajasthan, and its biocontrol potential against the soft rot pathogen Pectobacterium carotovorum subsp. carotovorum (Pcc). QQ activity was confirmed by the loss of violacein pigment in Chromobacterium violaceum (MCC 2290). Quorum quenching metabolites were extracted using ethyl acetate, and the Quorum Quenching Extract (QQE) demonstrated positive activity in assays with C. violaceum CV026 (MCC 2216). HRLC-MS analysis identified diketopiperazines, L,L-Cyclo (leucylprolyl) and Cyclo (L-Phe-L-Pro), which are N-acyl homoserine lactones (AHLs) antagonists competing for LuxR receptor binding sites. In vitro and in planta assays evaluated QQB's biocontrol potential using treatment I (Pcc), treatment II (Pcc + QQB), and a control (sterile water). In the in vitro soft rot attenuation assay showed that treatment I caused severe maceration in vegetable slices, particularly in radish, exhibiting the highest maceration diameter (25.33 ± 3.52 mm) and percentage (46.14 ± 5.70 %). However, co-inoculation with QQB significantly reduced maceration across all tested vegetables. In the plate assay, germination rates decreased to approximately 50 % in both Vigna radiata and Raphanus sativus for treatment I, but improved to 86.67 % in treatment II. The seed vigour and germination indices also improved with QQB treatment in both plant species. In the pot assay after 30 days, in contrast to a 50 % decrease in root and shoot lengths in treatment I, treatment II led to a substantial recovery, with root lengths increase by 112.07 % and 138.76 %, while shoot length by 315.65 % and 163.63 % in V. radiata and R. sativus, respectively. This study highlights the QQ and biocontrol potential of Bacillus sp. EBS9 against P. carotovorum (Pcc), suggesting its promise in effective management of phytopathogens, which is crucial for agricultural productivity while minimizing environmental impact.

Isolation of a quorum quenching bacterium effective to various acyl-homoserine lactones: Its quorum quenching mechanism and application to a membrane bioreactor

Biofouling, caused by microbial biofilm formation on the membrane surface and in pores, is a major operational problem in membrane bioreactors (MBR). Many quorum quenching (QQ) bacteria have been isolated and applied to MBR to reduce biofouling. However, for more effective MBR biofouling control, novel approaches for isolating QQ bacteria and applying them in MBR are needed. Therefore, Listeria grayi (HEMM-2) was isolated using a mixture of different N-acyl homoserine lactones (AHLs). HEMM-2 degraded various AHLs, regardless of the length and oxo group in the carbon chain, with quorum sensing (QS) inhibition ratios of 47–61%. This QQ activity was attributed to extracellular substances in HEMM-2 cell-free supernatant (CFS). Furthermore, the HEMM-2 CFS negatively regulated QS-related gene expression, inhibiting Pseudomonas aeruginosa and activated sludge-biofilm formation by 53–75%. Surprisingly, when the HEMM-2 CFS was directly injected into a laboratory-scale MBR system, biofouling was not significantly affected. Biofouling was only controlled by cell suspension (CS) of HEMM-2, indicating the importance of QQ bacteria in MBR. The HEMM-2 CS increased operation time to reach 0.4 bar, a threshold transmembrane pressure for complete biofouling, from 315 h to 371 h. Taken together, HEMM-2, which is effective in the degradation of various AHLs, and its applicable method to MBR may be considered a potent approach for controlling biofouling and understanding the behavior of QQ bacteria in MBR systems.

Inhibiting Biofilm Formation via Simultaneous Application of Nitric Oxide and Quorum Quenching Bacteria.

Membrane biofouling is an inevitable challenge in membrane-based water treatment systems such as membrane bioreactors. Recent studies have shown that biological approaches based on bacterial signaling can effectively control biofilm formation. Quorum quenching (QQ) is known to inhibit biofilm growth by disrupting quorum sensing (QS) signaling, while nitric oxide (NO) signaling helps to disperse biofilms. In this study, batch biofilm experiments were conducted to investigate the impact of simultaneously applying NO signaling and QQ for biofilm control using Pseudomonas aeruginosa PAO1 as a model microorganism. The NO treatment involved the injection of NONOates (NO donor compounds) into mature biofilms, while QQ was implemented by immobilizing QQ bacteria (Escherichia coli TOP10-AiiO or Rhodococcus sp. BH4) in alginate or polyvinyl alcohol/alginate beads to preserve the QQ activity. When QQ beads were applied together with (Z)-1-[N-(3-aminopropyl)-N-(n-propyl) amino]diazen-1-ium-1,2-diolate (PAPA NONOate), they achieved a 39.0% to 71.3% reduction in biofilm formation, which was substantially higher compared to their individual applications (16.0% to 54.4%). These findings highlight the significant potential of combining QQ and NO technologies for effective biofilm control across a variety of processes that require enhanced biofilm inhibition.

A low N-acyl-homoserine lactone-degrading Stentrophomonas sp. that could efficiently mitigate biofouling in a membrane bioreactor

Quorum quenching (QQ) is a promising and sustainable strategy for mitigating membrane biofouling in membrane bioreactors (MBRs). In this study, six QQ bacterial strains were isolated, and two of them, Bacillus sp. L (strain L) and Stenotrophomonas sp.Y6 (strain Y6), were further studied. Interestingly, strain L could degrade quorum-sensing signal molecules more efficiently than strain Y6. However, strain Y6 could better mitigate membrane biofouling in MBR, reducing the TMP from 31 kPa to 10 kPa, compared with 17 kPa achieved by L at the end of a 25-day MBR operation. This might be due to the presence of extracellular QQ enzymes in Y6 and the rapid adaptation of Y6 to the conditions within the MBR. In addition, Y6 did not affect the contents of COD, NH4+-N, total nitrogen, and total phosphorus in the effluent released by the MBR. These findings may reveal a new connection between the QQ activity of the bacterial cells and their ability to mitigate biofouling, a prospect that could enable the evaluation of efficient QQ candidates for practical application in wastewater treatment.

First insights into the Aurelia aurita transcriptome response upon manipulation of its microbiome.

The associated diverse microbiome contributes to the overall fitness of Aurelia aurita, particularly to asexual reproduction. However, how A. aurita maintains this specific microbiome or reacts to manipulations is unknown. In this report, the response of A. aurita to manipulations of its native microbiome was studied by a transcriptomics approach. Microbiome-manipulated polyps were generated by antibiotic treatment and challenging polyps with a non-native, native, and potentially pathogenic bacterium. Total RNA extraction followed by RNAseq resulted in over 155 million reads used for a de novo assembly. The transcriptome analysis showed that the antibiotic-induced change and resulting reduction of the microbiome significantly affected the host transcriptome, e.g., genes involved in processes related to immune response and defense mechanisms were highly upregulated. Similarly, manipulating the microbiome by challenging the polyp with a high load of bacteria (2 × 107 cells/polyp) resulted in induced transcription of apoptosis-, defense-, and immune response genes. A second focus was on host-derived quorum sensing interference as a potential defense strategy. Quorum Quenching (QQ) activities and the respective encoding QQ-ORFs of A. aurita were identified by functional screening a cDNA-based expression library generated in Escherichia coli. Corresponding sequences were identified in the transcriptome assembly. Moreover, gene expression analysis revealed differential expression of QQ genes depending on the treatment, strongly suggesting QQ as an additional defense strategy. Overall, this study allows first insights into A. aurita's response to manipulating its microbiome, thus paving the way for an in-depth analysis of the basal immune system and additional fundamental defense strategies.

Pharmacoinformatics-Based Approach for Uncovering the Quorum-Quenching Activity of Phytocompounds against the Oral Pathogen, Streptococcus mutans.

Streptococcus mutans, a gram-positive oral pathogen, is the primary causative agent of dental caries. Biofilm formation, a critical characteristic of S. mutans, is regulated by quorum sensing (QS). This study aimed to utilize pharmacoinformatics techniques to screen and identify effective phytochemicals that can target specific proteins involved in the quorum sensing pathway of S. mutans. A computational approach involving homology modeling, model validation, molecular docking, and molecular dynamics (MD) simulation was employed. The 3D structures of the quorum sensing target proteins, namely SecA, SMU1784c, OppC, YidC2, CiaR, SpaR, and LepC, were modeled using SWISS-MODEL and validated using a Ramachandran plot. Metabolites from Azadirachta indica (Neem), Morinda citrifolia (Noni), and Salvadora persica (Miswak) were docked against these proteins using AutoDockTools. MD simulations were conducted to assess stable interactions between the highest-scoring ligands and the target proteins. Additionally, the ADMET properties of the ligands were evaluated using SwissADME and pkCSM tools. The results demonstrated that campesterol, meliantrol, stigmasterol, isofucosterol, and ursolic acid exhibited the strongest binding affinity for CiaR, LepC, OppC, SpaR, and Yidc2, respectively. Furthermore, citrostadienol showed the highest binding affinity for both SMU1784c and SecA. Notably, specific amino acid residues, including ASP86, ARG182, ILE179, GLU143, ASP237, PRO101, and VAL84 from CiaR, LepC, OppC, SecA, SMU1784c, SpaR, and YidC2, respectively, exhibited significant interactions with their respective ligands. While the docking study indicated favorable binding energies, the MD simulations and ADMET studies underscored the substantial binding affinity and stability of the ligands with the target proteins. However, further in vitro studies are necessary to validate the efficacy of these top hits against S. mutans.

Potential role of quorum quenching activity of silver nanoparticles in controlling non-filamentous bulking within activated sludge process

The effective prevention and control of non-filamentous bulking is a significant challenge. In this study, the underlying effect of quorum sensing (QS) on inducing non-filamentous bulking and the maintenance effect of silver nanoparticles (AgNPs) on sludge floc stability, aggregation and settleability based on the quorum quenching (QQ) activity during non-filamentous bulking were investigated. The results showed that the concentration of N-acyl homoserine lactone (AHL) increased significantly in the activated sludge system at a high organic load rate (OLR), triggering the AHL-mediated QS. Additionally, the triggered QS promoted exopolysaccharide secretion, reducing the surface charge and hydrophobicity of the sludge aggregates, and further deteriorating the settleability of the sludge aggregates. AgNPs, a quorum sensing inhibitor (QSI), inhibited the AHL-QS based on QQ activity under high OLR, which maintained the physicochemical properties of extracellular polymeric substances (EPS). AgNPs-QQ maintained the surface energy barrier and electrostatic barrier of sludge aggregates and the gel properties of exopolysaccharides, which is favorable for microbial aggregation. The appropriate concentrations of AgNPs (≤10 mg/L) had no negative effect on biological nutrient removal in the sequencing batch reactors (SBRs) at the high organic loading. Therefore, AgNPs effectively prevent and control non-filamentous bulking by their QQ activity in the activated sludge process. Thus, the present study provided new insights into controlling non-filamentous bulking during the activated sludge process.

Anti-Inflammatory and Anti-Quorum Sensing Effect of Camellia sinensis Callus Lysate for Treatment of Acne.

Cutibacterium acnes (C. acnes) is involved in the pathogenesis of acne by inducing inflammation and biofilm formation, along with other virulence factors. A Camellia sinensis (C. sinensis) callus lysate is proposed to reduce these effects. The aim of the present work is to study the anti-inflammatory properties of a callus extract from C. sinensis on C. acnes-stimulated human keratinocytes and the quorum-quenching activities. Keratinocytes were stimulated with thermo-inactivated pathogenic C. acnes and were treated with the herbal lysate (0.25% w/w) to evaluate its anti-inflammatory effect. C. acnes biofilm was developed in vitro and treated with 2.5 and 5% w/w of the lysate to evaluate quorum sensing and the lipase activity. The results showed that the lysate was able to reduce the production of interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), and C-X-C motif chemokine ligand 1 (CXCL1), and decrease the nuclear translocation of nuclear factor kappa light chain enhancer of activated B cells (NF-κB). The lysate did not show bactericidal activity but showed diminished biofilm formation, the lipase activity, and the production of autoinducer 2 (AI-2), a member of a family of signaling molecules used in quorum sensing. Therefore, the proposed callus lysate could have the potential to reduce acne-related symptoms without the eradication of C. acnes, which is part of the natural skin microbiome.

Selection and Molecular Response of AHL-lactonase (aiiA) Producing Bacillus sp. Under Penicillin G-induced Conditions.

Quorum sensing (QS) is the process by which microorganisms employ chemicals called autoinducers (AIs) to communicate with their population. The QS mechanism generally controls the expression of the virulence related genes in bacteria. N-acyl homoserine lactones (AHLs) are the most widespread QS molecules. Due to their diverse AHL-lactonase activities, Bacillus species make particularly suitable candidates for procedures such as demolition of pathogenic bacterial QS signals and bioremediation of β-lactam antibiotics from contaminated environments. In this study, seven Bacillus strains with Quorum quenching (QQ) activity were isolated using an enrichment medium supplemented with Penicillin G (PenG). The AHL-lactonase encoding gene (aiiA) was amplified by PCR and sequenced. Amino acid sequences underwent multiple sequence alignment. Docking studies were carried out with both C6HSL and PenG ligand using AutoDock tools. The aiiA amino acid sequences of the isolates were found to be well conserved. Furthermore, amino acid sequence alignment revealed that 74.9% of amino acid sequences were conserved in the genus Bacillus. Docking of the C6HSL to wild type (3DHA) and H97D variant reduced the docking score by only 0.1kcal/mol for the mutated protein. When PenG docked with a higher (1.5kcal/mol) score as a ligand to wild-type and mutant receptors, the docking score for the mutated protein likewise decreased by 0.1kcal/mol. This research contributed to the diversification of organisms with QQ activity and beta-lactam antibiotic resistance. It also clarified the binding score of the PenG ligand to the Bacillus AHL lactonase molecule for the first time.

Effect of quorum quenching on biofouling control and microbial community in membrane bioreactors by Brucella sp. ZJ1

Quorum quenching (QQ) has been demonstrated to be a novel technique for controlling biofouling in membrane bioreactors (MBRs), as it can significantly inhibit biofilm formation by disrupting quorum sensing (QS). The exploration of new QQ bacterial strains and the evaluation of their performance in mitigating membrane fouling in MBR systems is significant. In this study, an efficient QQ strain, Brucella sp. ZJ1 was encapsulated in alginate beads and evaluated for its ability to mitigate biofouling. The findings revealed that MBR with QQ beads extended the operation time by 2–3 times without affecting pollutant degradation. QQ beads maintained approximately 50% QQ activity after more than 50 days operation, indicating a long-lasting and endurable QQ effect. The QQ effect reduced extracellular polymeric substance (EPS) production especially in terms of polysaccharide and protein by more than 40%. QQ beads in the MBR also reduced the cake resistance and the irreversible resistance of membrane biofouling. Metagenomic sequencing suggests that QQ beads suppressed the QS effect and increased the abundance of QQ enzyme genes, ultimately inducing efficient membrane biofouling control.

A method establishment and application for biofilm quorum quenching activity assay

The existence of quorum sensing (QS) and quorum quenching (QQ) plays important roles in biofilm formation. However, direct detection of QS ability is difficult due to the low concentrations of signal molecules inside the biofilm. Therefore, QQ activity is typically used to indicate the attribution of QS/QQ to the biofilm. Nevertheless, current detection methods of QQ activity based on biosensors present undesirable operability and accuracy. In this study, the 96-well plate assay based on a specific biosensor, Agrobacterium tumefaciens A136, and a colorimetric substance, X-gal was established. The reliable fitting results were obtained by standardizing the composition of the A136 X-gal assay solution and optimizing the operating conditions. This method improved the accuracy of QQ activity detection and reduced time and cost consumption. Finally, the 96-well plate assay was successfully applied to detect the QQ activities of biofilm samples and explore possible environmental influencing factors. In general, this study provided a new strategy for understanding the QQ effect in biofilm systems.