scholarly journals Disruption of Cell-to-Cell Signaling Does Not Abolish the Antagonism of Phaeobacter gallaeciensis toward the Fish Pathogen Vibrio anguillarum in Algal Systems

2013 ◽  
Vol 79 (17) ◽  
pp. 5414-5417 ◽  
Author(s):  
M. J. Prol García ◽  
P. W. D'Alvise ◽  
L. Gram
Keyword(s):  
Quorum Sensing ◽  
Cell Signaling ◽  
Disease Control ◽  
Vibrio Anguillarum ◽  
Fish Pathogen ◽  
Wild Type ◽  
Content Type ◽  
Phaeobacter Gallaeciensis ◽  
Algal Cultures

ABSTRACTQuorum sensing (QS) regulatesPhaeobacter gallaeciensisantagonism in broth systems; however, we demonstrate here that QS is not important for antagonism in algal cultures. QS mutants reducedVibrio anguillarumto the same extent as the wild type. Consequently, a combination of probioticPhaeobacterand QS inhibitors is a feasible strategy for aquaculture disease control.

scholarly journals Quorum Sensing Determines the Choice of Antiphage Defense Strategy in Vibrio anguillarum

mBio ◽  
2015 ◽  
Vol 6 (3) ◽  
Author(s):  
Demeng Tan ◽  
Sine Lo Svenningsen ◽  
Mathias Middelboe
Keyword(s):  
Quorum Sensing ◽  
Cell Density ◽  
Defense Mechanisms ◽  
Vibrio Anguillarum ◽  
Phage Infection ◽  
Phage Resistance ◽  
Fish Pathogen ◽  
Content Type ◽  
Density State ◽  
Infection Pressure

ABSTRACTSelection for phage resistance is a key driver of bacterial diversity and evolution, and phage-host interactions may therefore have strong influence on the genetic and functional dynamics of bacterial communities. In this study, we found that an important, but so far largely overlooked, determinant of the outcome of phage-bacterial encounters in the fish pathogenVibrio anguillarumis bacterial cell-cell communication, known as quorum sensing. Specifically,V. anguillarumPF430-3 cells locked in the low-cell-density state (ΔvanTmutant) express high levels of the phage receptor OmpK, resulting in a high susceptibility to phage KVP40, but achieve protection from infection by enhanced biofilm formation. By contrast, cells locked in the high-cell-density state (ΔvanΟmutant) are almost completely unsusceptible due to quorum-sensing-mediated downregulation of OmpK expression. The phenotypes of the two quorum-sensing mutant strains are accurately reflected in the behavior of wild-typeV. anguillarum, which (i) displays increased OmpK expression in aggregated cells compared to free-living variants in the same culture, (ii) displays a clear inverse correlation betweenompKmRNA levels and the concentration ofN-acylhomoserine lactone quorum-sensing signals in the culture medium, and (iii) survives mainly by one of these two defense mechanisms, rather than by genetic mutation to phage resistance. Taken together, our results demonstrate thatV. anguillarumemploys quorum-sensing information to choose between two complementary antiphage defense strategies. Further, the prevalence of nonmutational defense mechanisms in strain PF430-3 suggests highly flexible adaptations to KVP40 phage infection pressure, possibly allowing the long-term coexistence of phage and host.IMPORTANCEComprehensive knowledge on bacterial antiphage strategies and their regulation is essential for understanding the role of phages as drivers of bacterial evolution and diversity. In an applied context, development of successful phage-based control of bacterial pathogens also requires detailed understanding of the mechanisms of phage protection in pathogenic bacteria. Here, we demonstrate for the first time the presence of quorum-sensing-regulated phage defense mechanisms in the fish pathogenVibrio anguillarumand provide evidence that quorum-sensing regulation allowsV. anguillarumto alternate between different phage protection mechanisms depending on population cell density. Further, our results demonstrate the prevalence of nonmutational defense mechanisms in the investigatedV. anguillarumstrain, which allow flexible adaptations to a dynamic phage infection pressure.

scholarly journals Quorum Sensing Influences Burkholderia thailandensis Biofilm Development and Matrix Production

2016 ◽  
Vol 198 (19) ◽  
pp. 2643-2650 ◽  
Author(s):  
Boo Shan Tseng ◽  
Charlotte D. Majerczyk ◽  
Daniel Passos da Silva ◽  
Josephine R. Chandler ◽  
E. Peter Greenberg ◽  
...  
Keyword(s):  
Quorum Sensing ◽  
Biofilm Formation ◽  
Bacterial Species ◽  
Matrix Material ◽  
Close Relative ◽  
Wild Type ◽  
Flow Conditions ◽  
Burkholderia Thailandensis ◽  
Content Type ◽  
Biofilm Development

ABSTRACTMembers of the genusBurkholderiaare known to be adept at biofilm formation, which presumably assists in the survival of these organisms in the environment and the host. Biofilm formation has been linked to quorum sensing (QS) in several bacterial species. In this study, we characterizedBurkholderia thailandensisbiofilm development under flow conditions and sought to determine whether QS contributes to this process.B. thailandensisbiofilm formation exhibited an unusual pattern: the cells formed small aggregates and then proceeded to produce mature biofilms characterized by “dome” structures filled with biofilm matrix material. We showed that this process was dependent on QS.B. thailandensishas three acyl-homoserine lactone (AHL) QS systems (QS-1, QS-2, and QS-3). An AHL-negative strain produced biofilms consisting of cell aggregates but lacking the matrix-filled dome structures. This phenotype was rescued via exogenous addition of the three AHL signals. Of the threeB. thailandensisQS systems, we show that QS-1 is required for proper biofilm development, since abtaR1mutant, which is defective in QS-1 regulation, forms biofilms without these dome structures. Furthermore, our data show that the wild-type biofilm biomass, as well as the material inside the domes, stains with a fucose-binding lectin. ThebtaR1mutant biofilms, however, are negative for fucose staining. This suggests that the QS-1 system regulates the production of a fucose-containing exopolysaccharide in wild-type biofilms. Finally, we present data showing that QS ability during biofilm development produces a biofilm that is resistant to dispersion under stress conditions.IMPORTANCEThe saprophyteBurkholderia thailandensisis a close relative of the pathogenic bacteriumBurkholderia pseudomallei, the causative agent of melioidosis, which is contracted from its environmental reservoir. Since most bacteria in the environment reside in biofilms,B. thailandensisis an ideal model organism for investigating questions inBurkholderiaphysiology. In this study, we characterizedB. thailandensisbiofilm development and sought to determine if quorum sensing (QS) contributes to this process. Our work shows thatB. thailandensisproduces biofilms with unusual dome structures under flow conditions. Our findings suggest that these dome structures are filled with a QS-regulated, fucose-containing exopolysaccharide that may be involved in the resilience ofB. thailandensisbiofilms against changes in the nutritional environment.

scholarly journals Quorum Sensing Signal Selectivity and the Potential for Interspecies Cross Talk

mBio ◽  
2019 ◽  
Vol 10 (2) ◽  
Author(s):  
Samantha Wellington ◽  
E. Peter Greenberg
Keyword(s):  
Quorum Sensing ◽  
Cell Signaling ◽  
Cross Talk ◽  
Homoserine Lactone ◽  
Interspecies Interactions ◽  
Interspecies Competition ◽  
Content Type ◽  
E Coli ◽  
Competition And Cooperation ◽  
Wide Range

ABSTRACTMany species of proteobacteria communicate with kin and coordinate group behaviors through a form of cell-cell signaling called acyl-homoserine lactone (AHL) quorum sensing (QS). Most AHL receptors are thought to be specific for their cognate signal, ensuring that bacteria cooperate and share resources only with closely related kin cells. Although specificity is considered fundamental to QS, there are reports of “promiscuous” receptors that respond broadly to nonself signals. These promiscuous responses expand the function of QS systems to include interspecies interactions and have been implicated in both interspecies competition and cooperation. Because bacteria are frequently members of polymicrobial communities, AHL cross talk between species could have profound impacts. To better understand the prevalence of QS promiscuity, we measured the activity of seven QS receptors in their native host organisms. To facilitate comparison of our results to previous studies, we also measured receptor activity using heterologous expression inEscherichia coli. We found that the standardE. colimethods consistently overestimate receptor promiscuity and sensitivity and that overexpression of the receptors is sufficient to account for the discrepancy between native andE. colireporters. Additionally, receptor overexpression resulted in AHL-independent activity inPseudomonas aeruginosa. Using our activation data, we developed a quantitative score of receptor selectivity. We find that the receptors display a wide range of selectivity and that most receptors respond sensitively and strongly to at least one nonself signal, suggesting a broad potential for cross talk between QS systems.IMPORTANCESpecific recognition of cognate signals is considered fundamental to cell signaling circuits as it creates fidelity in the communication system. In bacterial quorum sensing (QS), receptor specificity ensures that bacteria cooperate only with kin. There are examples, however, of QS receptors that respond promiscuously to multiple signals. “Eavesdropping” by these promiscuous receptors can be beneficial in both interspecies competition and cooperation. Despite their potential significance, we know little about the prevalence of promiscuous QS receptors. Further, many studies rely on methods requiring receptor overexpression, which is known to increase apparent promiscuity. By systematically studying QS receptors in their natural parent strains, we find that the receptors display a wide range of selectivity and that there is potential for significant cross talk between QS systems. Our results provide a basis for hypotheses about the evolution and function of promiscuous signal receptors and for predictions about interspecies interactions in complex microbial communities.

scholarly journals Phenylacetyl Coenzyme A, Not Phenylacetic Acid, Attenuates CepIR-Regulated Virulence in Burkholderia cenocepacia

2019 ◽  
Vol 85 (24) ◽  
Author(s):  
Tasia Joy Lightly ◽  
Kara L. Frejuk ◽  
Marie-Christine Groleau ◽  
Laurent R. Chiarelli ◽  
Cor Ras ◽  
...  
Keyword(s):  
Quorum Sensing ◽  
Phenylacetic Acid ◽  
Coenzyme A ◽  
Opportunistic Pathogen ◽  
Burkholderia Cenocepacia ◽  
Signal Molecules ◽  
Wild Type ◽  
Content Type ◽  
Opportunistic Bacteria ◽  
Virulent Phenotype

ABSTRACT During phenylalanine catabolism, phenylacetic acid (PAA) is converted to phenylacetyl coenzyme A (PAA-CoA) by a ligase, PaaK, and then PAA-CoA is epoxidized by a multicomponent monooxygenase, PaaABCDE, before further degradation through the tricarboxylic acid (TCA) cycle. In the opportunistic pathogen Burkholderia cenocepacia, loss of paaABCDE attenuates virulence factor expression, which is under the control of the LuxIR-like quorum sensing (QS) system, CepIR. To further investigate the link between CepIR-regulated virulence and PAA catabolism, we created knockout mutants of the first step of the pathway (PAA-CoA synthesis by PaaK) and characterized them in comparison to a paaABCDE mutant using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and virulence assays. We found that while loss of PaaABCDE decreased virulence, deletion of the paaK genes resulted in a more virulent phenotype than that of the wild-type strain. Deletion of either paaK or paaABCDE led to higher levels of released PAA but no differences in levels of internal accumulation compared to the wild-type level. While we found no evidence of direct cepIR downregulation by PAA-CoA or PAA, a low-virulence cepR mutant reverted to a virulent phenotype upon removal of the paaK genes. On the other hand, removal of paaABCDE in the cepR mutant did not impact its attenuated phenotype. Together, our results suggest an indirect role for PAA-CoA in suppressing B. cenocepacia CepIR-activated virulence. IMPORTANCE The opportunistic pathogen Burkholderia cenocepacia uses a chemical signal process called quorum sensing (QS) to produce virulence factors. In B. cenocepacia, QS relies on the presence of the transcriptional regulator CepR which, upon binding QS signal molecules, activates virulence. In this work, we found that even in the absence of CepR, B. cenocepacia can elicit a pathogenic response if phenylacetyl-CoA, an intermediate of the phenylacetic acid degradation pathway, is not produced. Instead, accumulation of phenylacetyl-CoA appears to attenuate pathogenicity. Therefore, we have discovered that it is possible to trigger virulence in the absence of CepR, challenging the classical view of activation of virulence by this QS mechanism. Our work provides new insight into the relationship between metabolism and virulence in opportunistic bacteria. We propose that in the event that QS signaling molecules cannot accumulate to trigger a pathogenic response, a metabolic signal can still activate virulence in B. cenocepacia.

scholarly journals Quorum Sensing Is Accompanied by Global Metabolic Changes in the Opportunistic Human Pathogen Pseudomonas aeruginosa

2015 ◽  
Vol 197 (12) ◽  
pp. 2072-2082 ◽  
Author(s):  
Peter W. Davenport ◽  
Julian L. Griffin ◽  
Martin Welch
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Fatty Acid ◽  
Quorum Sensing ◽  
Acid Methyl Ester ◽  
Metabolic Changes ◽  
Human Pathogen ◽  
Wild Type ◽  
Content Type ◽  
Ecological Fitness ◽  
Opportunistic Human Pathogen

ABSTRACTPseudomonas aeruginosausesN-acyl-homoserine lactone (AHL)-dependent quorum sensing (QS) systems to control the expression of secreted effectors. These effectors can be crucial to the ecological fitness of the bacterium, playing roles in nutrient acquisition, microbial competition, and virulence. In this study, we investigated the metabolic consequences of AHL-dependent QS by monitoring the metabolic profile(s) of alasI rhlIdouble mutant (unable to make QS signaling molecules) and its wild-type progenitor as they progressed through the growth curve. Analysis of culture supernatants by1H-nuclear magnetic resonance (1H-NMR) spectroscopy revealed that at the point where AHL concentrations peaked in the wild type, the metabolic footprints (i.e., extracellular metabolites) of the wild-type andlasI rhlImutant diverged. Subsequent gas chromatography-mass spectrometry (GC-MS)-based analysis of the intracellular metabolome revealed QS-dependent perturbations in around one-third of all identified metabolites, including altered concentrations of tricarboxylic acid (TCA) cycle intermediates, amino acids, and fatty acids. Further targeted fatty acid methyl ester (FAME) GC-MS-based profiling of the cellular total fatty acid pools revealed that QS leads to changes associated with decreased membrane fluidity and higher chemical stability. However, not all of the changes we observed were necessarily a direct consequence of QS; liquid chromatography (LC)-MS analyses revealed that polyamine levels were elevated in thelasI rhlImutant, perhaps a response to the absence of QS-dependent adaptations. Our data suggest that QS leads to a global readjustment in central metabolism and provide new insight into the metabolic changes associated with QS during stationary-phase adaptation.IMPORTANCEQuorum sensing (QS) is a transcriptional regulatory mechanism that allows bacteria to coordinate their gene expression profile with the population cell density. The opportunistic human pathogenPseudomonas aeruginosauses QS to control the production of secreted virulence factors. In this study, we show that QS elicits a global “metabolic rewiring” inP. aeruginosa. This metabolic rerouting of fluxes is consistent with a variety of drivers, ranging from altered QS-dependent transcription of “metabolic genes” through to the effect(s) of global “metabolic readjustment” as a consequence of QS-dependent exoproduct synthesis, as well as a general stress response, among others. To our knowledge, this is the first study of its kind to assess the global impact of QS on the metabolome.

scholarly journals The CpAL Quorum Sensing System Regulates Production of Hemolysins CPA and PFO To Build Clostridium perfringens Biofilms

2015 ◽  
Vol 83 (6) ◽  
pp. 2430-2442 ◽  
Author(s):  
Jorge E. Vidal ◽  
Joshua R. Shak ◽  
Adrian Canizalez-Roman
Keyword(s):  
Quorum Sensing ◽  
Clostridium Perfringens ◽  
Biofilm Formation ◽  
Extracellular Dna ◽  
Wild Type ◽  
Content Type ◽  
Type C ◽  
Enteritis Necroticans ◽  
Biofilm Structure

Clostridium perfringensstrains produce severe diseases, including myonecrosis and enteritis necroticans, in humans and animals. Diseases are mediated by the production of potent toxins that often damage the site of infection, e.g., skin epithelium during myonecrosis. In planktonic cultures, the regulation of important toxins, such as CPA, CPB, and PFO, is controlled by theC. perfringensAgr-like (CpAL) quorum sensing (QS) system. Strains also encode a functional LuxS/AI-2 system. AlthoughC. perfringensstrains form biofilm-like structures, the regulation of biofilm formation is poorly understood. Therefore, our studies investigated the role of CpAL and LuxS/AI-2 QS systems and of QS-regulated factors in controlling the formation of biofilms. We first demonstrate that biofilm production by reference strains differs depending on the culture medium. Increased biomass correlated with the presence of extracellular DNA in the supernatant, which was released by lysis of a fraction of the biofilm population and planktonic cells. Whereas ΔagrBmutant strains were not able to produce biofilms, a ΔluxSmutant produced wild-type levels. The transcript levels of CpAL-regulatedcpaandpfoAgenes, but notcpb, were upregulated in biofilms compared to planktonic cultures. Accordingly, Δcpaand ΔpfoAmutants, in type A (S13) or type C (CN3685) backgrounds, were unable to produce biofilms, whereas CN3685Δcpbmade wild-type levels. Biofilm formation was restored in complemented Δcpa/cpaand ΔpfoA/pfoAstrains. Confocal microscopy studies further detected CPA partially colocalizing with eDNA on the biofilm structure. Thus, CpAL regulates biofilm formation inC. perfringensby increasing levels of certain toxins required to build biofilms.

scholarly journals The Type IX Secretion System Is Required for Virulence of the Fish Pathogen Flavobacterium psychrophilum

2020 ◽  
Vol 86 (16) ◽  
Author(s):  
Paul Barbier ◽  
Tatiana Rochat ◽  
Haitham H. Mohammed ◽  
Gregory D. Wiens ◽  
Jean-François Bernardet ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Gene Deletion ◽  
Cold Water ◽  
Secretion System ◽  
Gliding Motility ◽  
Control Measures ◽  
Fish Pathogen ◽  
Wild Type ◽  
Content Type ◽  
Bacterial Cold Water Disease

ABSTRACT Flavobacterium psychrophilum causes bacterial cold-water disease in wild and aquaculture-reared fish and is a major problem for salmonid aquaculture. The mechanisms responsible for cold-water disease are not known. It was recently demonstrated that the related fish pathogen, Flavobacterium columnare, requires a functional type IX protein secretion system (T9SS) to cause disease. T9SSs secrete cell surface adhesins, gliding motility proteins, peptidases, and other enzymes, any of which may be virulence factors. The F. psychrophilum genome has genes predicted to encode components of a T9SS. Here, we used a SacB-mediated gene deletion technique recently adapted for use in the Bacteroidetes to delete a core F. psychrophilum T9SS gene, gldN. The ΔgldN mutant cells were deficient for secretion of many proteins in comparison to wild-type cells. Complementation of the mutant with wild-type gldN on a plasmid restored secretion. Compared to wild-type and complemented strains, the ΔgldN mutant was deficient in adhesion, gliding motility, and extracellular proteolytic and hemolytic activities. The ΔgldN mutant exhibited reduced virulence in rainbow trout and complementation restored virulence, suggesting that the T9SS plays an important role in the disease. IMPORTANCE Bacterial cold-water disease, caused by F. psychrophilum, is a major problem for salmonid aquaculture. Little is known regarding the virulence factors involved in this disease, and control measures are inadequate. A targeted gene deletion method was adapted to F. psychrophilum and used to demonstrate the importance of the T9SS in virulence. Proteins secreted by this system are likely virulence factors and targets for the development of control measures.

scholarly journals Vibrio anguillarum Is Genetically and Phenotypically Unaffected by Long-Term Continuous Exposure to the Antibacterial Compound Tropodithietic Acid

2016 ◽  
Vol 82 (15) ◽  
pp. 4802-4810 ◽  
Author(s):  
Bastian Barker Rasmussen ◽  
Torben Grotkjær ◽  
Paul W. D'Alvise ◽  
Guangliang Yin ◽  
Faxing Zhang ◽  
...  
Keyword(s):  
Food Production ◽  
Fish Larvae ◽  
Vibrio Anguillarum ◽  
Resistant Bacteria ◽  
Continuous Exposure ◽  
Fish Pathogen ◽  
Content Type ◽  
Tropodithietic Acid ◽  
Antibacterial Compound

ABSTRACTMinimizing the use of antibiotics in the food production chain is essential for limiting the development and spread of antibiotic-resistant bacteria. One alternative intervention strategy is the use of probiotic bacteria, and bacteria of the marineRoseobacterclade are capable of antagonizing fish-pathogenic vibrios in fish larvae and live feed cultures for fish larvae. The antibacterial compound tropodithietic acid (TDA), an antiporter that disrupts the proton motive force, is key in the antibacterial activity of several roseobacters. Introducing probiotics on a larger scale requires understanding of any potential side effects of long-term exposure of the pathogen to the probionts or any compounds they produce. Here we exposed the fish pathogenVibrio anguillarumto TDA for several hundred generations in an adaptive evolution experiment. No tolerance or resistance arose during the 90 days of exposure, and whole-genome sequencing of TDA-exposed lineages and clones revealed few mutational changes, compared to lineages grown without TDA. Amino acid-changing mutations were found in two to six different genes per clone; however, no mutations appeared unique to the TDA-exposed lineages or clones. None of the virulence genes ofV. anguillarumwas affected, and infectivity assays using fish cell lines indicated that the TDA-exposed lineages and clones were less invasive than the wild-type strain. Thus, long-term TDA exposure does not appear to result in TDA resistance and the physiology ofV. anguillarumappears unaffected, supporting the application of TDA-producing roseobacters as probiotics in aquaculture.IMPORTANCEIt is important to limit the use of antibiotics in our food production, to reduce the risk of bacteria developing antibiotic resistance. We showed previously that marine bacteria of theRoseobacterclade can prevent or reduce bacterial diseases in fish larvae, acting as probiotics. Roseobacters produce the antimicrobial compound tropodithietic acid (TDA), and we were concerned regarding whether long-term exposure to this compound could induce resistance or affect the disease-causing ability of the fish pathogen. Therefore, we exposed the fish pathogenVibrio anguillarumto increasing TDA concentrations over 3 months. We did not see the development of any resistance to TDA, and subsequent infection assays revealed that none of the TDA-exposed clones had increased virulence toward fish cells. Hence, this study supports the use of roseobacters as a non-risk-based disease control measure in aquaculture.

scholarly journals The Quorum Sensing Regulator CinR Hierarchically Regulates Two Other Quorum Sensing Pathways in Ligand-Dependent and -Independent Fashions in Rhizobium etli

2015 ◽  
Vol 197 (9) ◽  
pp. 1573-1581 ◽  
Author(s):  
Huiming Zheng ◽  
Yiling Mao ◽  
Qingcheng Zhu ◽  
Jun Ling ◽  
Na Zhang ◽  
...  
Keyword(s):  
Quorum Sensing ◽  
Homoserine Lactone ◽  
Nodule Formation ◽  
Rhizobium Etli ◽  
Wild Type ◽  
Content Type ◽  
Nitrogen Fixation Activity ◽  
Regulatory Cascade ◽  
Regulatory Systems ◽  
Cell Densities

ABSTRACTMany rhizobial species use complexN-acyl-homoserine lactone (AHL)-based quorum sensing (QS) systems to monitor their population density and regulate their symbiotic interactions with their plant hosts. There are at least three LuxRI-type regulatory systems inRhizobium etliCFN42: CinRI, RaiRI, and TraRI. In this study, we show that CinI, RaiI, and TraI are responsible for synthesizing all AHLs under the tested conditions. The activation of these AHL synthase genes requires their corresponding LuxR-type counterparts. We further demonstrate that CinRI is at the top of the regulatory cascade that activates RaiRI and TraRI QS systems. Moreover, we discovered that CinR possesses a specific affinity to bindcinIpromoter in the absence of its cognate AHL ligand, thereby activatingcinItranscription. Addition of AHLs leads to improved binding to thecinIpromoter and enhancedcinIexpression. Furthermore, we found that compared to the wild type, thecinRmutation displayed reduced nodule formation, andcinR,raiR, andtraImutants show significantly lower levels of nitrogen fixation activity than the wild type. These results suggest that the complex QS regulatory systems inR. etliplay an important role in its symbiosis with legume hosts.IMPORTANCEMany bacteria use quorum sensing (QS) to monitor their cell densities and coordinately regulate a number of physiological functions. Rhizobia often have diverse and complex LuxR/LuxI-type quorum sensing systems that may be involved in symbiosis and N2fixation. In this study, we identified three LuxR/LuxI-type QS systems inRhizobium etliCFN42: CinRI, RaiRI, and TraRI. We established a complex network of regulation between these QS components and found that these QS systems played important roles in symbiosis processes.

scholarly journals Genetic Analysis of the Upper Phenylacetate Catabolic Pathway in the Production of Tropodithietic Acid by Phaeobacter gallaeciensis

2012 ◽  
Vol 78 (10) ◽  
pp. 3539-3551 ◽  
Author(s):  
Martine Berger ◽  
Nelson L. Brock ◽  
Heiko Liesegang ◽  
Marco Dogs ◽  
Ines Preuth ◽  
...  
Keyword(s):  
Aerobic Bacteria ◽  
Insertion Mutagenesis ◽  
Wild Type ◽  
Quantitative Real Time Pcr ◽  
Catabolic Pathway ◽  
Phylogenetic Groups ◽  
Content Type ◽  
Transposon Insertion ◽  
Tropodithietic Acid ◽  
Phaeobacter Gallaeciensis

ABSTRACTProduction of the antibiotic tropodithietic acid (TDA) depends on the central phenylacetate catabolic pathway, specifically on the oxygenase PaaABCDE, which catalyzes epoxidation of phenylacetyl-coenzyme A (CoA). Our study was focused on genes of the upper part of this pathway leading to phenylacetyl-CoA as precursor for TDA.Phaeobacter gallaeciensisDSM 17395 encodes two genes with homology to phenylacetyl-CoA ligases (paaK1andpaaK2), which were shown to be essential for phenylacetate catabolism but not for TDA biosynthesis and phenylalanine degradation. Thus, inP. gallaeciensisanother enzyme must produce phenylacetyl-CoA from phenylalanine. Using random transposon insertion mutagenesis of apaaK1-paaK2double mutant we identified a gene (ior1) with similarity toiorAandiorBin archaea, encoding an indolepyruvate:ferredoxin oxidoreductase (IOR). Theior1mutant was unable to grow on phenylalanine, and production of TDA was significantly reduced compared to the wild-type level (60%). Nuclear magnetic resonance (NMR) spectroscopic investigations using13C-labeled phenylalanine isotopomers demonstrated that phenylalanine is transformed into phenylacetyl-CoA by Ior1. Using quantitative real-time PCR, we could show that expression ofior1depends on the adjacent regulator IorR. Growth on phenylalanine promotes production of TDA, induces expression ofior1(27-fold) andpaaK1(61-fold), and regulates the production of TDA. Phylogenetic analysis showed that the aerobic type of IOR as found in many roseobacters is common within a number of different phylogenetic groups of aerobic bacteria such asBurkholderia,Cupriavidis, andRhizobia, where it may also contribute to the degradation of phenylalanine.

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