La Sr Research Articles

Identification of a solo acylhomoserine lactone synthase from the myxobacterium Archangium gephyra

Considered a key taxon in soil and marine microbial communities, myxobacteria exist as coordinated swarms that utilize a combination of lytic enzymes and specialized metabolites to facilitate predation of microbes. This capacity to produce specialized metabolites and the associated abundance of biosynthetic pathways contained within their genomes have motivated continued drug discovery efforts from myxobacteria. Of all myxobacterial biosynthetic gene clusters deposited in the antiSMASH database, only one putative acylhomoserine lactone (AHL) synthase, agpI, was observed, in genome data from Archangium gephyra. Without an AHL receptor also apparent in the genome of A. gephyra, we sought to determine if AgpI was an uncommon example of an orphaned AHL synthase. Herein we report the bioinformatic assessment of AgpI and discovery of a second AHL synthase from Vitiosangium sp. During axenic cultivation conditions, no detectible AHL metabolites were observed in A. gephyra extracts. However, heterologous expression of each synthase in Escherichia coli provided detectible quantities of 3 AHL signals including 2 known AHLs, C8-AHL and C9-AHL. These results suggest that A. gephyra AHL production is dormant during axenic cultivation. The functional, orphaned AHL synthase, AgpI, is unique to A. gephyra, and its utility to the predatory myxobacterium remains unknown.

Extremely relative cooling power of Cu0.35Zn0.65Fe2O4

Here, it is reported that magnetocaloric effect of Cu0.35Zn0.65Fe2O4 (CZF) ferrite covers a large range of temperature as an effective material for magnetic refrigerator (MR). In addition, the relative cooling power (RCP) in CZF ferrite is significantly larger than corresponding values of the famous magnetic refrigerants such as Gd, MnAs1−xSbx, Gd5Si2Ge2, (La–Sr)MnO3, (Pr–Sr)MnO3, (La–Sr)MnO3, (La–Ca–Pb)MnO3, and (La–Ca–Sr)MnO3. Therefore, CZF ferrite is useful to be a working substance in MR as a single magnetic refrigerant, including room temperature and very low temperatures.

Co-dependent and Interdigitated: Dual Quorum Sensing Systems Regulate Conjugative Transfer of the Ti Plasmid and the At Megaplasmid in Agrobacterium tumefaciens 15955.

Members of the Rhizobiaceae, often carry multiple secondary replicons in addition to the primary chromosome with compatible repABC-based replication systems. Unlike secondary chromosomes and chromids, repABC-based megaplasmids and plasmids can undergo copy number fluctuations and are capable of conjugative transfer in response to environmental signals. Several Agrobacterium tumefaciens lineages harbor three secondary repABC-based replicons, including a secondary chromosome (often linear), the Ti (tumor-inducing) plasmid and the At megaplasmid. The Ti plasmid is required for virulence and encodes a conjugative transfer (tra) system that is strictly regulated by a subset of plant-tumor released opines and a well-described acyl-homoserine lactone (AHL)-based quorum-sensing mechanism. The At plasmids are generally not required for virulence, but carry genes that enhance rhizosphere survival, and these plasmids are often conjugatively proficient. We report that the At megaplasmid of the octopine-type strain A. tumefaciens 15955 encodes a quorum-controlled conjugation system that directly interacts with the paralogous quorum sensing system on the co-resident Ti plasmid. Both the pAt15955 and pTi15955 plasmids carry homologs of a TraI-type AHL synthase, a TraR-type AHL-responsive transcription activator, and a TraM-type anti-activator. The traI genes from both pTi15955 and pAt15955 can direct production of the inducing AHL (3-octanoyl-L-homoserine lactone) and together contribute to the overall AHL pool. The TraR protein encoded on each plasmid activates AHL-responsive transcription of target tra gene promoters. The pAt15955 TraR can cross-activate tra genes on the Ti plasmid as strongly as its cognate tra genes, whereas the pTi15955 TraR is preferentially biased toward its own tra genes. Putative tra box elements are located upstream of target promoters, and comparing between plasmids, they are in similar locations and share an inverted repeat structure, but have distinct consensus sequences. The two AHL quorum sensing systems have a combinatorial effect on conjugative transfer of both plasmids. Overall, the interactions described here have implications for the horizontal transfer and evolutionary stability of both plasmids and, in a broad sense, are consistent with other repABC systems that often have multiple quorum-sensing controlled secondary replicons.

The Investigation of Left Atrial Structure and Stroke Etiology: The I-LASER Study.

BackgroundLeft atrial (LA) function is important in stroke, but often poorly characterized. We evaluated the association of 2‐dimensional speckle tracking echocardiography LA variables with stroke subtype (cardioembolic stroke [CS] or cryptogenic stroke versus other). The hypothesis is worse LA active function is associated with CS, but not cryptogenic strokes.Methods and ResultsIn this prospective cohort (2017–2019), left ventricular/LA structure and function were quantified by 2‐dimensional and speckle tracking echocardiography in 151 patients with stroke. Strain/strain rate curves for the 3 components of the LA cycle, ie, (1) Reservoir (global longitudinal strain [Srmax]), (2) Conductive (early LA Sr [Sre]), and (3) Active (late LA strain [Sra]) were evaluated, masked to stroke subtype. Associations of cardiac features with stroke subtype were tested using multivariable logistic regressions. Odds of CS were increased in patients with a larger LA systolic diameter (odds ratio [OR], 2.96, 95% CI, 1.14–7.69) but reduced in patients with a higher Srmax (better reservoir) (OR, 0.80, 95% CI, 0.67–0.97). Lower Sra (worse function) was associated with an increased odds of CS (OR, 1.72, 95% CI, 1.07–2.76) but not independent of atrial fibrillation. Higher active LA emptying fraction (better active phase) was associated with reduced odds of CS (OR, 0.74, 95% CI, 0.57–0.95) or cryptogenic stroke (OR, 0.82, 95% CI, 0.68–0.98) versus other subtypes; other associations between cryptogenic stroke and speckle tracking echocardiography were not found.ConclusionsMarkers of LA structure and function were associated with CS. Similar associations were not found for cryptogenic stroke, which might suggest different underlying mechanisms, given study limitations. Further understanding could aid stroke diagnosis and secondary stroke prevention research.

Oxygen availability affects the synthesis of quorum sensing signal in the facultative anaerobe Novosphingobium pentaromativorans US6-1.

Bacterial populations rely on quorum sensing (QS) to coordinate their behaviors and are often challenged by the fluctuation in oxygen concentrations in their habitats. Oxygen is a crucial factor that affects bacterial metabolism in multiple ways. However, little is known about whether and how oxygen availability affects QS activities. To fill this gap, we used the facultative anaerobe Novosphingobium pentaromativorans US6-1 as a model system, and observed that the QS signal acyl homoserine-lactones (AHLs) were produced only in anoxic environments, such as biofilm, or liquid medium that initially contained less than 2 mg/L dissolved oxygen, but not in highly oxic environments. Comparative transcriptome analysis revealed that oxygen availability significantly affected the physiological activities in US6-1, including fatty acid metabolism, oxidative phosphorylation, citrate cycle, QS activities, and flagellar assembly. The absence of AHLs in the oxic culture was not due to degradation, but to the very low expression of the AHL synthase gene novI. High concentration of NADH during the middle log phase under static cultivation may be a trigger for AHL synthesis. This is the first report that production of AHLs is coupled with anoxic metabolism in a facultative anaerobe, which extends our knowledge on factors affecting bacterial QS occurrence. KEY POINTS: • AHL production is anoxic cultivation related. • Oxygen availability affects AHL synthesis by influencing novI expression. • Oxygen availability changes many metabolism activities including NADH production.

Bacterial quorum sensing quenching activity of Lysobacter leucyl aminopeptidase acts by interacting with autoinducer synthase

Acyl-homoserine lactone (AHL) is the most studied autoinducer in gram-negative bacteria controlling infections of various pathogens. Quenching of AHL signaling by inhibiting AHL synthesis or AHL-receptor binding via small molecular chemicals or enzymatically degrading AHL is commonly used to block bacterial infections. Here, we describe a new quorum-quenching strategy that directly “acquires” bacterial genes/proteins through a defined platform. We artificially expressed a typical AHL synthase gene pcoI from the biocontrol Pseudomonas fluorescens 2P24 in the antifungal bacterium Lysobacter enzymogenes OH11 lacking AHL production. This step led to the discovery of multiple PcoI interacting protein candidates from L. enzymogenes. The individual expression of these candidate genes in 2P24 led to the identification of Le0959, which encodes leucyl aminopeptidase, an effective protein that inhibits AHL synthesis in 2P24. Therefore, we define Le0959 as LqqP (Lysobacterquorum-quenching protein). The expression of pcoI in E. coli could produce AHL, and the introduction of lqqP into E. coli expressing pcoI could prevent the production of AHL. LqqP directly binds to PcoI, and this protein–protein binding reduced the abundance of free PcoI (capable of AHL synthesis) in vivo, thereby blocking PcoI-dependent AHL production. Overall, this study highlights the discovery of LqqP in quenching AHL quorum sensing by binding to AHL synthase via developing a previously-uncharacterized screening technique for bacterial quorum quenching.

Sr doping effect on the structure property and NO oxidation performance of dual-site doped perovskite La(Sr)Co(Fe)O3

In this work, the physico-chemical properties of the La1-xSrxCo0.8Fe0.2O3 (x = 0, 0.3，0.4，0.5，0.6) catalysts were systematically studied as well as their catalytic activity for NO oxidation. When Sr2+ substituted at La3+ site, [CoO6] octahedral distortion is amended and the crystal structure transited from rhombohedral to cubic. Co(III) electronic configuration turned from high/intermediate spin state to low spin sate with empty eg filling. Imbalanced positive charge induced by Sr2+ was compensated by tetravalent Co(IV) ions and oxygen deficiencies together. All these factors resulted in better reducibility of Co, increasing oxygen deficiencies and more mobile oxygen species. Thus, NO conversion of 67–69% was attained over the Sr-doped catalysts at 300 °C, much higher than the undoped catalysts (NO conversion of 24.3%). A significant decrease of the apparent activation energy (Ea), from 68 kJ/mol down to ~52 kJ/mol, was observed as well. Based on the kinetics results, NO oxidation over the perovskite catalysts is a typical superficial process determined by adsorbed NO and surface oxygen species.

Quorum alleviating Molecules- A Review

The hindrance of quorum sensing is generally indicated to as quorum quenching. A few compounds have been combined to intrude on quorum sensing articulation by echoing AHL particles, N-acyl-3-amino-5H furanone, a synthetic analogue of N-AHL viably keeps the receipt of signal atom by obstructing its receptor Lux R protein. AHL controlled quorum sensing can be distressed by various methods, for example, lessening the movement of AHL cognate receptor protein or AHL synthase, hindering the creation of QS signal molecules, degradation of the AHL, and mimicking the signal atoms by utilizing their synthetic analogs. Additionally, obstruction of sensing signal transduction can be accomplished by utilizing an opponent particle equipped for contending or meddling with the local AHL motion for endorsed entry to the LUXR-type receptor. Competitive inhibitors are basically like the local AHL signal and can bind to and involve the AHL restricting site yet will fail to enact the LuxR-type receptor. The non-competitive inhibitors indicate almost no basic similarity to AHL signals and these particles bind to various locales on the receptor protein. A variety of natural Quorum quenching molecules have been identified.

(Invited) Effects of Cobalt Co-Doping on Mixed Ion Conductivity in La(Sr)Ga(Fe)O3 for Oxygen Separation

Mixed ion conductor shows oxide ion and electronic conduction simultaneously and expecting for application to oxygen separation membrane from air. Because of significant volume change depending on oxygen partial pressure, oxygen separation membrane using mixed ion conductor has disadvantage of narrow range of oxygen partial pressure resulting in the limited permeation rate. Therefore, increased oxide ion conductivity and mechanical strength have been required for mixed ion conducting membrane for oxygen separator. In our previous study, we found that Fe doped LaGaO3 shows reasonably high oxide ion conductivity and chemical stability. In particular, La0.7Sr0.3Ga0.6Fe0.4O3 (LSGF) shows largest oxide permeation rate from air to He. In this study, for further increase in oxygen permeation rate, effects of dopant to Ga site in LSGF was investigated. Application to CH4 partial oxidation was also studied. All LSGF sample was prepared with conventional solid state reaction method using oxide as source materials. Partial substitution was performed for Ga site in LSGF and dense sample without impurity phase was confirmed by XRD measurement. For oxygen permeation measurement, La0.6Sr0.4CoO3 catalyst was coated on the surface of LSGF disk (Φ17mm diameter, 0.5mm thickness) It was found that the oxygen permeation from air to He was sensitively influenced by dopant and the oxygen permeation rate was increased as the following order, Co>Al>In>Ni>Mn at 1273 K. On the other hand, in case of Ru and Ir which is easily reduced to metallic state decreased the oxygen permeation rate significantly. Therefore, it was found that addition of 10 mol % of Co or Al is effective for increasing oxygen permeation rate of LSGF. Effects of Co in LSGF was further studied from oxygen permeation rate and the highest oxygen permeation rate was achieved at 3 mol% in Ga site of LSGC. On this optimized composition, La0.7Sr0.3Ga0.57Fe0.4Co0.03O3, shows 225 μmol min-1 cm-2 at 1273 K and this value is almost 2.5 times larger than that of LSGF. Effects of small amount of Co on electrical conductivity in LSGF were also studied. Although LSGF shows high hole conduction like log(σ/Scm-1)=0.6, the conductivity was increased by Co doping, in particular, at lower temperature. In contrast, Al doping decreases total conductivity and so, increase in oxygen permeation rate could be assigned to the increased mobility of oxide ion in LSGF. Application of Co doped LSGF to CH4 partial oxidation was further studied. Although optimized amount of Co doped for Ga site in LSGF is 3 mol%, the membrane was broken when 3 mol% doped LSGF was used for oxygen membrane. 1% Co doped LSGF was applied for oxygen permeation membrane and it was found that amount of oxygen permeation rate was 1.5 times increased comparing with LSGF. In addition, no crack was formed in membrane. The amount of oxygen permeation was achieved 364μmol cm2 min-1 which is corresponded to 8.9ml/min cm2 at 1273K. In summary, this study reveals that doping small amount of Co to Ga site of LSGF is highly effective for increasing oxygen permeation rate without decreasing chemical stability.

Pd and GDC Co-infiltrated LSCM cathode for high-temperature CO2 electrolysis using solid oxide electrolysis cells

• Pd-GDC co-infiltrated LSCM cathode for the high-temperature CO 2 electrolysis was invesigated. • Pd-GDC@LSCM material presented highly dispersed Pd-GDC nanoparticles on LSCM. • This structure extended the triple-phase boundaries to enhance the electrochemical activity. • Using the Pd-GDC@LSCM electrode, CO 2 was reduced at a rate of 2362 μmol cm −2 min −1 at 1123 K. • It is possible to achieve preliminary performance through the Pd-GDC nanoparticles onto the LSCM. The electrochemical reduction of CO 2 using a highly efficient solid oxide electrolyzer could be considered an alternative to the advanced utilization of CO 2 . The La(Sr)Cr(Mn)O 3 (LSCM) perovskite oxide has previously been examined as a promising ceramic cathode material for application in a CO 2 solid oxide electrolyzer at high temperatures. However, LSCM suffers from low electrocatalytic activity towards CO 2 reduction. In this study, a modified LSCM-based cathode material is fabricated by co-infiltrating Pd metal and Ce 0.8 Gd 0.2 O 1.9 (GDC) nanoparticles on the surface of the LSCM scaffold. Structural characterization and electrochemical analysis of the high-temperature CO 2 electrolysis procedure are conducted for various CO/CO 2 mixtures and at different operating temperatures. The enhanced electrocatalytic activity of the Pd-GDC co-infiltrated LSCM cathode compared to LSCM is attributed to the increased numbers of active triple phase boundaries and surface oxygen vacancies resulting from the co-infiltration of Pd-GDC nanoparticles on the LSCM cathode.

PcsR2 Is a LuxR-Type Regulator That Is Upregulated on Wheat Roots and Is Unique to Pseudomonas chlororaphis.

LuxR solos are common in plant-associated bacteria and increasingly recognized for playing important roles in plant-microbe interkingdom signaling. Unlike the LuxR-type transcriptional regulators of prototype LuxR/LuxI quorum sensing systems, luxR solos do not have a LuxI-type autoinducer synthase gene associated with them. LuxR solos in plant-pathogenic bacteria are important for virulence and in plant endosymbionts contribute to symbiosis. In the present study, we characterized an atypical LuxR solo, PcsR2, in the biological control species Pseudomonas chlororaphis 30–84 that is highly conserved among sequenced P. chlororaphis strains. Unlike most LuxR solos in the plant-associated bacteria characterized to date, pcsR2 is not associated with a proline iminopeptidase gene and the protein has an atypical N-terminal binding domain. We created a pcsR2 deletion mutant and used quantitative RT-PCR to show that the expression of pcsR2 and genes in the operon immediately downstream was upregulated ∼10-fold when the wild type strain was grown on wheat roots compared to planktonic culture. PcsR2 was involved in upregulation. Using a GFP transcriptional reporter, we found that expression of pcsR2 responded specifically to root-derived substrates as compared to leaf-derived substrates but not to endogenous AHLs. Compared to the wild type, the mutant was impaired in the ability to utilize root carbon and nitrogen sources in wheat root macerate and to colonize wheat roots. Phenazine production and most biofilm traits previously shown to be correlated with phenazine production also were diminished in the mutant. Gene expression of several of the proteins in the phenazine regulatory network including PhzR, Pip (phenazine inducing protein) and RpeA/RpeB were reduced in the mutant, and overexpression of these genes in trans restored phenazine production in the mutant to wild-type levels, indicating PcsR2 affects the activity of the these regulatory genes upstream of RpeA/RpeB via an undetermined mechanism. Our results indicate PcsR2 upregulates the expression of the adjacent operon in response to unknown wheat root-derived signals and belongs to a novel subfamily of LuxR-type transcriptional regulators found in sequenced P. chlororaphis strains.

Stringent starvation protein A and LuxI/LuxR-type quorum sensing system constitute a mutual positive regulation loop in Pseudoalteromonas

Bacteria commonly exhibit social activities through acyl-homoserine lactones (AHLs)-based quorum sensing (QS) systems to form their unique social network. The sigma factor RpoS is an important regulator that controls QS system in different bacteria. However, the upstream of RpoS involving regulation on QS system remains unclear. In Escherichia coli RpoS is regulated by stringent starvation protein A (SspA), which is dependent of histone-like nucleoid structuring protein (H-NS). To date, the connection between SspA and QS system is essentially unknown. Here, we characterized a typical LuxI/LuxR-type QS system in marine bacterium Pseudoalteromonas sp. T1lg65 which can produce four types of AHLs. The luxI encoding AHLs synthase and luxR encoding AHLs-responsive receptor are co-transcribed, providing advantages in rapidly amplifying QS signaling. Notably, SspA positively regulated luxI/luxR transcription by activating RpoS expression, which is mediated by H-NS. Interestingly, LuxR in turn positively regulated SspA expression. Therefore, SspA and QS system constitute a mutual positive regulation loop in T1lg65. In view of the crucial roles of SspA and QS system in environmental adaption, we believe that the improvement of bacterial tolerance to marine environments could be related to rapidly tuning SspA-involved QS programming.

AbaR is a LuxR type regulator essential for motility and the formation of biofilm and pellicle in Acinetobacter baumannii.

Acinetobacter baumannii is a major opportunistic pathogen causing nosocomial infections. Acinetobacterbaumannii possesses a quorum sensing system consisting of abaI, encoding an autoinducer synthase, and abaR, encoding a putative LuxR type regulator. AbaI is required for motility and biofilm formation in A. baumannii. However, the functions of AbaR on the expression of abaI, motility, and the formation of biofilm and pellicle have not yet been explored. The aim of this study was to investigate the effects of abaR mutation on the expression of abaI, motility, and the formation of biofilm and pellicle. Functions of AbaR were assessed by the construction of an isogenic mutant and by evaluating the effects of abaR mutation on the expression of abaI, motility, and the formation of biofilm and pellicle. The abaR mutant revealed a significant decrease in the expression of abaI. The disruption of abaR resulted in substantial defects in motility and the formation of biofilm and pellicle. Introduction of abaR in trans complemented the defects. AbaR of A. baumannii is required for the expression of abaI and plays important roles in motility and the formation of biofilm and pellicle. AbaR may be considered to be a target of anti-biofilm agents.

Quorum Sensing as a Target for Controlling Surface Associated Motility and Biofilm Formation in Acinetobacter baumannii ATCC® 17978TM.

The important nosocomial pathogen Acinetobacter baumannii presents a quorum sensing (QS) system (abaI/abaR) mediated by acyl-homoserine-lactones (AHLs) and several quorum quenching (QQ) enzymes. However, the roles of this complex network in the control of the expression of important virulence-related phenotypes such as surface-associated motility and biofilm formation is not clear. Therefore, the effect of the mutation of the AHL synthase AbaI, and the exogenous addition of the QQ enzyme Aii20J on surface-associated motility and biofilm formation by A. baumannii ATCC® 17978TM was studied in detail. The effect of the enzyme on biofilm formation by several multidrug-resistant A. baumannii clinical isolates differing in their motility pattern was also tested. We provide evidence that a functional QS system is required for surface-associated motility and robust biofilm formation in A. baumannii ATCC® 17978TM. Important differences were found with the well-studied strain A. nosocomialis M2 regarding the relevance of the QS system depending on environmental conditions The in vitro biofilm-formation capacity of A. baumannii clinical strains was highly variable and was not related to the antibiotic resistance or surface-associated motility profiles. A high variability was also found in the sensitivity of the clinical strains to the action of the QQ enzyme, revealing important differences in virulence regulation between A. baumannii isolates and confirming that studies restricted to a single strain are not representative for the development of novel antimicrobial strategies. Extracellular DNA emerges as a key component of the extracellular matrix in A. baumannii biofilms since the combined action of the QQ enzyme Aii20J and DNase reduced biofilm formation in all tested strains. Results demonstrate that QQ strategies in combination with other enzymatic treatments such as DNase could represent an alternative approach for the prevention of A. baumannii colonization and survival on surfaces and the prevention and treatment of infections caused by this pathogen.

Role of plant compounds in the modulation of the conjugative transfer of pRet42a.

One of the most studied mechanisms involved in bacterial evolution and diversification is conjugative transfer (CT) of plasmids. Plasmids able to transfer by CT often encode beneficial traits for bacterial survival under specific environmental conditions. Rhizobium etli CFN42 is a Gram-negative bacterium of agricultural relevance due to its symbiotic association with Phaseolus vulgaris through the formation of Nitrogen-fixing nodules. The genome of R. etli CFN42 consists of one chromosome and six large plasmids. Among these, pRet42a has been identified as a conjugative plasmid. The expression of the transfer genes is regulated by a quorum sensing (QS) system that includes a traI gene, which encodes an acyl-homoserine lactone (AHL) synthase and two transcriptional regulators (TraR and CinR). Recently, we have shown that pRet42a can perform CT on the root surface and inside nodules. The aim of this work was to determine the role of plant-related compounds in the CT of pRet42a. We found that bean root exudates or root and nodule extracts induce the CT of pRet42a in the plant rhizosphere. One possibility is that these compounds are used as nutrients, allowing the bacteria to increase their growth rate and reach the population density leading to the activation of the QS system in a shorter time. We tested if P. vulgaris compounds could substitute the bacterial AHL synthesized by TraI, to activate the conjugation machinery. The results showed that the transfer of pRet42a in the presence of the plant is dependent on the bacterial QS system, which cannot be substituted by plant compounds. Additionally, individual compounds of the plant exudates were evaluated; among these, some increased and others decreased the CT. With these results, we suggest that the plant could participate at different levels to modulate the CT, and that some compounds could be activating genes in the conjugation machinery.

Topotactic phase transformations by concerted dual-ion migration of B-site cation and oxygen in multivalent cobaltite La–Sr–Co–Ox films

Manipulating topotactic phase transformations via orderly ion transfer in complex oxides ABO x is ubiquitous in advanced applications such as ionotronics, ion-batteries and catalysts. Most of such ion-mediated transformations are accomplished by the transfer of oxygen or A-site ions. However, implementing the transformation via the transfer of B-site ions, despite the great challenge to overcome a large cohesive energy, has unique advantage since they host most functional properties of materials. Here, we present a tri-state phase transformation from perovskite (P) to brownmillerite (BM) and to single-layered perovskite (SL) structure via the concerted migration of oxygen and B-site Co-ions in La 0.7 Sr 0.3 CoO 3 thin films. Ac-STEM, XPS, XAS, PNR, magnetic and electric measurements demonstrated that presented B-site Co-cation transfer is along the CoO 4 tetrahedral sub-layer of the BM film, which leads to the reconfiguration of 3 d -electrons and spin state in remanent Co ions and causes tremendous changes in magnetic and electric properties: from canted-antiferromagnetic insulator in BM phase to ferromagnetic insulator in SL phase. First-principles calculations revealed that the La 3+ -doping at A-site largely reduces the cohesive energy of Co-ions in CoO 4 and destabilize the CoO 4 tetrahedron of BM phase, which explains the formation of Co-ions transfer channel in the CoO 4 tetrahedral sub-layer. The present study highlights the effectiveness of regulating topotactic transformation via B-site ions transfer and provides a new pathway for manipulating the topotactic transformation with diverse functionalities. Tri-state topotactic transformation induced by B-site Co ion transfer was firstly realized in La 0.7 Sr 0.3 CoO 3 film. A unique transfer channel for Co-ions is demonstrated to be facilitated by the instability of CoO 4 due to A-site La-doping. The reconfiguration of 3d-electrons due to Co transfer leads to tremendous changes in magnetic and electric properties. Such new topotactic transformation driven by B-site ion transfer offers opportunity to exploit new types of electrocatalyst with rich functionalities. • Tri-state topotactic transformation induced by B-site Co ion transfer was firstly realized in La 0.7 Sr 0.3 CoO 3 film. • A unique transfer channel for Co-ions is demonstrated to be facilitated by the instability of CoO 4 due to A-site La-doping. • The concerted transfer of Co and O ions drive tremendous changes in magnetic and electric properties. • Such B-site ion transfer provides a new pathway for exploiting electrocatalyst with diverse functionalities.

Regulation of the AcrAB efflux system by the quorum-sensing regulator AnoR in Acinetobacter nosocomialis.

Multidrug efflux pumps play an important role in antimicrobial resistance and pathogenicity in bacteria. Here, we report the functional characterization of the RND (resistance-nodulation- division) efflux pump, AcrAB, in Acinetobacter nosocomialis. An in silico analysis revealed that homologues of the AcrAB efflux pump, comprising AcrA and AcrB, are widely distributed among different bacterial species. Deletion of acrA and/or acrB genes led to decreased biofilm/pellicle formation and reduced antimicrobial resistance in A. nosocomialis. RNA sequencing and mRNA expression analyses showed that expression of acrA/B was downregulated in a quorum sensing (QS) regulator (anoR)-deletion mutant, indicating transcriptional activation of the acrAB operon by AnoR in A. nosocomialis. Bioassays showed that secretion of N-acyl homoserine lactones (AHLs) was unaffected in acrA and acrB deletion mutants; however, AHL secretion was limited in a deletion mutant of acrR, encoding the acrAB regulator, AcrR. An in silico analysis indicated the presence of AcrR-binding motifs in promoter regions of anoI (encoding AHL synthase) and anoR. Specific binding of AcrR was confirmed by electrophoretic mobility shift assays, which revealed that AcrR binds to positions -214 and -217 bp upstream of the translational start sites of anoI and anoR, respectively, demonstrating transcriptional regulation of these QS genes by AcrR. The current study further addresses the possibility that AcrAB is controlled by the osmotic stress regulator, OmpR, in A. nosocomialis. Our data demonstrate that the AcrAB efflux pump plays a crucial role in biofilm/pellicle formation and antimicrobial resistance in A. nosocomialis, and is under the transcriptional control of a number of regulators. In addition, the study emphasizes the interrelationship of QS and AcrAB efflux systems in A. nosocomialis.

Direct Binding of Salicylic Acid to Pectobacterium N-Acyl-Homoserine Lactone Synthase.

Salicylic acid (SA) is a hormone that mediates systemic acquired resistance in plants. We demonstrated that SA can interfere with group behavior and virulence of the soft-rot plant pathogen Pectobacterium spp. through quorum sensing (QS) inhibition. QS is a population density-dependent communication system that relies on the signal molecule acyl-homoserine lactone (AHL) to synchronize infection. P. parmentieri mutants, lacking the QS AHL synthase (expI-) or the response regulator (expR-), were used to determine how SA inhibits QS. ExpI was expressed in DH5α, the QS negative strain of Escherichia coli, revealing direct interference of SA with AHL synthesis. Docking simulations showed SA is a potential ExpI ligand. This hypothesis was further confirmed by direct binding of SA to purified ExpI, shown by isothermal titration calorimetry and microscale thermophoresis. Computational alanine scanning was employed to design a mutant ExpI with predicted weaker binding affinity to SA. The mutant was constructed and displayed lower affinity to the ligand in the binding assay, and its physiological inhibition by SA was reduced. Taken together, these data support a likely mode of action and a role for SA as potent inhibitor of AHL synthase and QS.

Structure-Guided Biochemical Analysis of Quorum Signal Synthase Specificities.

Many bacteria use membrane-diffusible small molecule quorum signals to coordinate gene transcription in response to changes in cell density, known as quorum sensing (QS). Among these, acyl-homoserine lactones (AHL) are widely distributed in Proteobacteria and are involved in controlling the expression of virulence genes and biofilm formation in pathogens, such as Pseudomonas aeruginosa. AHL molecules are specifically biosynthesized by the cognate LuxI type AHL synthases using S-adenosylmethionine (SAM) and either acyl carrier protein (ACP)- or CoA-coupled fatty acids through a two-step reaction. Here, we characterize a CoA-dependent LuxI synthase from Rhodopseudomonas palustris that utilizes an aryl-CoA substrate that is environmentally derived, specifically p-coumaric acid. We leverage structures of this aryl-CoA-dependent synthase, along with our prior studies of an acyl-CoA-dependent synthase, to identify residues that confer substrate chain specificity in these enzymes. We test our predictions by carrying out biochemical, kinetic, and structural characterization of representative AHL signal synthases. Our studies provide an understanding of various AHL synthases that may be deployed in synthetic biological applications and inform on the design of specific small molecule therapeutics that can restrict virulence by targeting quorum signaling.

Cladodionen Is a Potential Quorum Sensing Inhibitor Against Pseudomonas aeruginosa.

Pseudomonas aeruginosa is an opportunistic pathogen using virulence factors and biofilm regulated by quorum sensing (QS) systems to infect patients and protect itself from environmental stress and antibiotics. Interfering with QS systems is a novel approach to combat P. aeruginosa infections without killing the bacteria, meaning that it is much harder for bacteria to develop drug resistance. A marine fungus Cladosporium sp. Z148 with anti-QS activity was obtained from Jiaozhou Bay, China. Cladodionen, a novel QS inhibitor, was isolated from the extracts of this fungus. Cladodionen had a better inhibitory effect than pyocyanin on the production of elastase and rhamnolipid. It also inhibited biofilm formation and motilities. The mRNA expressions of QS-related genes, including receptor proteins (lasR, rhlR and pqsR), autoinducer synthases (lasI, rhlI and pqsA) and virulence factors (lasB and rhlA) were down-regulated by cladodionen. Molecular docking analysis showed that cladodionen had better binding affinity to LasR and PqsR than natural ligands. Moreover, the binding affinity of cladodionen to LasR was higher than to PqsR. Cladodionen exhibits potential as a QS inhibitor against P. aeruginosa, and its structure–activity relationships should be further studied to illustrate the mode of action, optimize its structure and improve anti-QS activity.