Production and Degradation ofN-Acylhomoserine Lactone Quorum Sensing Signal Molecules in Bacteria Isolated from Activated Sludge

2013 ◽  
Vol 77 (12) ◽  
pp. 2436-2440 ◽  
Author(s):  
Seiji OCHIAI ◽  
Tomohiro MOROHOSHI ◽  
Ayane KURABEISHI ◽  
Masahiro SHINOZAKI ◽  
Haruka FUJITA ◽  
...  
Keyword(s):  
Quorum Sensing ◽  
Activated Sludge ◽  
Signal Molecules ◽  
Acylhomoserine Lactone

scholarly journals AmiE, a NovelN-Acylhomoserine Lactone Acylase Belonging to the Amidase Family, from the Activated-Sludge Isolate Acinetobacter sp. Strain Ooi24

2014 ◽  
Vol 80 (22) ◽  
pp. 6919-6925 ◽  
Author(s):  
Seiji Ochiai ◽  
Sera Yasumoto ◽  
Tomohiro Morohoshi ◽  
Tsukasa Ikeda
Keyword(s):  
Quorum Sensing ◽  
Activated Sludge ◽  
High Performance ◽  
Genomic Library ◽  
Amide Bond ◽  
Signal Molecules ◽  
Content Type ◽  
Chromatography Analysis ◽  
Acylhomoserine Lactone ◽  
Acyl Chains

ABSTRACTMany Gram-negative bacteria useN-acyl-l-homoserine lactones (AHLs) as quorum-sensing signal molecules. We have reported thatAcinetobacterstrains isolated from activated sludge have AHL-degrading activity. In this study, we cloned theamiEgene as an AHL-degradative gene from the genomic library ofAcinetobactersp. strain Ooi24. High-performance liquid chromatography analysis revealed that AmiE functions as an AHL acylase, which hydrolyzes the amide bond of AHL. AmiE showed a high level of degrading activity against AHLs with long acyl chains but no activity against AHLs with acyl chains shorter than eight carbons. AmiE showed homology with a member of the amidases (EC 3.5.1.4) but not with any known AHL acylase enzymes. An amino acid sequence of AmiE from Ooi24 showed greater than 99% identities with uncharacterized proteins fromAcinetobacter ursingiiCIP 107286 andAcinetobactersp. strain CIP 102129, but it was not found in the draft or complete genome sequences of otherAcinetobacterstrains. The presence of transposase-like genes around theamiEgenes of these threeAcinetobacterstrains suggests thatamiEis transferred by a putative transposon. Furthermore, the expression of AmiE inPseudomonas aeruginosaPAO1 reduced AHL accumulation and elastase activity, which were regulated by AHL-mediated quorum sensing.

scholarly journals Interference with Pseudomonas aeruginosa Quorum Sensing and Virulence by the Mycobacterial Pseudomonas Quinolone Signal Dioxygenase AqdC in Combination with the N-Acylhomoserine Lactone Lactonase QsdA

2019 ◽  
Vol 87 (10) ◽  
Author(s):  
Franziska S. Birmes ◽  
Ruth Säring ◽  
Miriam C. Hauke ◽  
Niklas H. Ritzmann ◽  
Steffen L. Drees ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Quorum Sensing ◽  
Virulence Factors ◽  
Rhodococcus Erythropolis ◽  
Quorum Quenching ◽  
Signal Molecules ◽  
Content Type ◽  
Acylhomoserine Lactone ◽  
Regulatory Effects ◽  
Epithelial Lung Cells

ABSTRACT The nosocomial pathogen Pseudomonas aeruginosa regulates its virulence via a complex quorum sensing network, which, besides N-acylhomoserine lactones, includes the alkylquinolone signal molecules 2-heptyl-3-hydroxy-4(1H)-quinolone (Pseudomonas quinolone signal [PQS]) and 2-heptyl-4(1H)-quinolone (HHQ). Mycobacteroides abscessus subsp. abscessus, an emerging pathogen, is capable of degrading the PQS and also HHQ. Here, we show that although M. abscessus subsp. abscessus reduced PQS levels in coculture with P. aeruginosa PAO1, this did not suffice for quenching the production of the virulence factors pyocyanin, pyoverdine, and rhamnolipids. However, the levels of these virulence factors were reduced in cocultures of P. aeruginosa PAO1 with recombinant M. abscessus subsp. massiliense overexpressing the PQS dioxygenase gene aqdC of M. abscessus subsp. abscessus, corroborating the potential of AqdC as a quorum quenching enzyme. When added extracellularly to P. aeruginosa cultures, AqdC quenched alkylquinolone and pyocyanin production but induced an increase in elastase levels. When supplementing P. aeruginosa cultures with QsdA, an enzyme from Rhodococcus erythropolis which inactivates N-acylhomoserine lactone signals, rhamnolipid and elastase levels were quenched, but HHQ and pyocyanin synthesis was promoted. Thus, single quorum quenching enzymes, targeting individual circuits within a complex quorum sensing network, may also elicit undesirable regulatory effects. Supernatants of P. aeruginosa cultures grown in the presence of AqdC, QsdA, or both enzymes were less cytotoxic to human epithelial lung cells than supernatants of untreated cultures. Furthermore, the combination of both aqdC and qsdA in P. aeruginosa resulted in a decline of Caenorhabditis elegans mortality under P. aeruginosa exposure.

scholarly journals Identification of Quorum-Sensing Signal Molecules and a Biosynthetic Gene in Alicycliphilus sp. Isolated from Activated Sludge

Sensors ◽  
2016 ◽  
Vol 16 (8) ◽  
pp. 1218 ◽  
Author(s):  
Tomohiro Morohoshi ◽  
Noriya Okutsu ◽  
Xiaonan Xie ◽  
Tsukasa Ikeda
Keyword(s):  
Quorum Sensing ◽  
Activated Sludge ◽  
Signal Molecules ◽  
Biosynthetic Gene

scholarly journals Advancing the Quorum in Pseudomonas aeruginosa: MvaT and the Regulation of N-Acylhomoserine Lactone Production and Virulence Gene Expression

2002 ◽  
Vol 184 (10) ◽  
pp. 2576-2586 ◽  
Author(s):  
Stephen P. Diggle ◽  
Klaus Winzer ◽  
Andrée Lazdunski ◽  
Paul Williams ◽  
Miguel Cámara
Keyword(s):  
Gene Expression ◽  
Pseudomonas Aeruginosa ◽  
Quorum Sensing ◽  
Growth Phase ◽  
Virulence Gene ◽  
Homoserine Lactone ◽  
Logarithmic Phase ◽  
Signal Molecules ◽  
Virulence Gene Expression ◽  
Acylhomoserine Lactone

ABSTRACT Pseudomonas aeruginosa regulates the production of many exoproteins and secondary metabolites via a hierarchical quorum-sensing cascade through LasR and RhlR and their cognate signal molecules N-(3-oxododecanoyl)-l-homoserine lactone (3O-C12-HSL) and N-(butanoyl)-l-homoserine lactone (C4-HSL). In this study, we found that transcription of the quorum sensing-regulated genes lecA (coding for PA-IL lectin), lasB (coding for elastase), and rpoS appeared to be growth phase dependent and their expression could not be advanced to the logarithmic phase in cells growing in batch culture by the addition of exogenous C4-HSL and 3O-C12-HSL. To identify novel regulators responsible for this growth phase dependency, a P. aeruginosa lecA::lux reporter strain was subjected to random transposon mutagenesis. A number of mutants affected in lecA expression were found that exhibited altered production of multiple quorum sensing-dependent phenotypes. While some mutations were mapped to new loci such as clpA and mvaT and a putative efflux system, a number of mutations were also mapped to known regulators such as lasR, rhlR, and rpoS. MvaT was identified as a novel global regulator of virulence gene expression, as a mutation in mvaT resulted in enhanced lecA expression and pyocyanin production. This mutant also showed altered swarming ability and production of the LasB and LasA proteases, 3O-C12-HSL, and C4-HSL. Furthermore, addition of exogenous 3O-C12-HSL and C4-HSL to the mvaT mutant significantly advanced lecA expression, suggesting that MvaT is involved in the growth phase-dependent regulation of the lecA gene.

N-Acylhomoserine lactone quorum-sensing signalling in antagonistic phenazine-producing Pseudomonas isolates from the red cocoyam rhizosphere

Microbiology ◽  
2011 ◽  
Vol 157 (2) ◽  
pp. 459-472 ◽  
Author(s):  
K. De Maeyer ◽  
J. D'aes ◽  
G. K. H. Hua ◽  
M. Perneel ◽  
L. Vanhaecke ◽  
...  
Keyword(s):  
Quorum Sensing ◽  
Regulatory System ◽  
Homoserine Lactone ◽  
Antagonistic Activity ◽  
Signal Molecules ◽  
Pseudomonas Chlororaphis ◽  
Homoserine Lactones ◽  
Acylhomoserine Lactone ◽  
Phenazine Production ◽  
Phenazine Antibiotics

Forty fluorescent Pseudomonas strains isolated from white and red cocoyam roots were tested for their ability to synthesize N-acyl-l-homoserine lactones (acyl-HSLs). Remarkably, only isolates from the red cocoyam rhizosphere that were antagonistic against the cocoyam root rot pathogen Pythium myriotylum and synthesized phenazine antibiotics produced acyl-HSLs. This supports the assumption that acyl-HSL production is related to the antagonistic activity of the strains. After detection, the signal molecules were identified through TLC-overlay and liquid chromatography-multiple MS (LC-MS/MS) analysis. In our representative strain, Pseudomonas CMR12a, production of the signal molecules could be assigned to two quorum-sensing (QS) systems. The first one is the QS system for phenazine production, PhzI/PhzR, which seemed to be well conserved, since it was genetically organized in the same way as in the well-described phenazine-producing Pseudomonas strains Pseudomonas fluorescens 2-79, Pseudomonas chlororaphis PCL1391 and Pseudomonas aureofaciens 30-84. The newly characterized genes cmrI and cmrR make up the second QS system of CMR12a, under the control of the uncommon N-3-hydroxy-dodecanoyl-homoserine lactone (3-OH-C12-HSL) and with low similarity to other Pseudomonas QS systems. No clear function could yet be assigned to the CmrI/CmrR system, although it contributes to the biocontrol capability of CMR12a. Both the PhzI/PhzR and CmrI/CmrR systems are controlled by the GacS/GacA two-component regulatory system.

Quorum Sensing in Burkholderia cepacia: Identification of the LuxRI Homologs CepRI

1999 ◽  
Vol 181 (3) ◽  
pp. 748-756 ◽  
Author(s):  
Shawn Lewenza ◽  
Barbara Conway ◽  
E. P. Greenberg ◽  
Pamela A. Sokol
Keyword(s):  
Quorum Sensing ◽  
Burkholderia Cepacia ◽  
Brain Heart Infusion ◽  
Homoserine Lactone ◽  
Signal Molecules ◽  
Acylhomoserine Lactone ◽  
Transposon Insertion ◽  
A Cell ◽  
Negative Role ◽  
Insertion Mutants

ABSTRACT Burkholderia cepacia has emerged as an important pathogen in patients with cystic fibrosis. Many gram-negative pathogens regulate the production of extracellular virulence factors by a cell density-dependent mechanism termed quorum sensing, which involves production of diffusible N-acylated homoserine lactone signal molecules, called autoinducers. Transposon insertion mutants of B. cepacia K56-2 which hyperproduced siderophores on chrome azurol S agar were identified. One mutant, K56-R2, contained an insertion in a luxR homolog that was designated cepR. The flanking DNA region was used to clone the wild-type copy of cepR. Sequence analysis revealed the presence of cepI, a luxI homolog, located 727 bp upstream and divergently transcribed from cepR. Alux box-like sequence was identified upstream ofcepI. CepR was 36% identical to Pseudomonas aeruginosa RhlR and 67% identical to SolR of Ralstonia solanacearum. CepI was 38% identical to RhlI and 64% identical to SolI. K56-R2 demonstrated a 67% increase in the production of the siderophore ornibactin, was protease negative on dialyzed brain heart infusion milk agar, and produced 45% less lipase activity in comparison to the parental strain. Complementation of acepR mutation restored parental levels of ornibactin and protease but not lipase. An N-acylhomoserine lactone was purified from culture fluids and identified asN-octanoylhomoserine lactone. K56-I2, a cepImutant, was created and shown not to produceN-octanoylhomoserine lactone. K56-I2 hyperproduced ornibactin and did not produce protease. These data suggest both a positive and negative role for cepIR in the regulation of extracellular virulence factor production by B. cepacia.

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