• Title/Summary/Keyword: AHL

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A Liquid Culture Bioassay System for the Detection of Quorum Sensing Signaling AHL Analogues

  • Kim, Young-Hee;Lee, Jae-Geun;Park, Sung-Hoon;Kim, Jung-Sun
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.224.1-224.1
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    • 2003
  • Recent studies have revealed that bacterial biofilm production by the gram-negative bacteria is regulated by the quorum sensing signaling molecules, AHLs (N-acyl homoserine lactones). This suggests that inhibiting the AHLs could enhance the effects of antibacterial agents. Halogenated furanones purified from the red algae Delisea pulchra have been known to decrease quorum sensing responses by competitive inhibition of the AHLs. (omitted)

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Rhizoctonia Blight of Azolla japonica Caused by Rhizoctonia solani (Rhizoctonia solani에 의한 큰물개구리밥(Azolla japonica) 마름병)

  • Lee, Jung-Han;Cha, Jea-Yul;Noh, Gil-Han;Han, Ki-Soo;Bae, Dong-Won;Kwon, Young-Sang;Lim, Chae-Shin;Jeong, Sung-Woo;Kwon, Jin-Hyeuk;Park, Chung-Gyoo;Kwak, Youn-Sig
    • Research in Plant Disease
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    • v.17 no.3
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    • pp.405-409
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    • 2011
  • Azolla Lam. is a small aquatic fern with deeply bilobed leaves, which are consisted of a thick greenish, with chlorophyll, upper (dorsal) lobe and a thinner, translucent lower (ventral) lobe, without chlorophyll, submerged in the water. Azolla blight was observed at a lotus pond. Mycological characteristics of the fungus associated with Azolla blight was immediately determined as Rhizoctonia sp. by the thickness and branching of hypha at right angles at the point toward the distal end of septa, with branching hypha is constricted. The fungus produced brown mycelia and dark brown sclerotia on PDA. The optimum temperature for mycelial growth and sclerotia formation were $25^{\circ}C$ and $30^{\circ}C$, respectively. The optimum temperature for fungal infection was $30^{\circ}C$, when spray inoculated. Phylogenetic analysis of rDNA-ITS revealed that the fungus was identified as Rhizoctonia solani (AG-1 IA) closest to one causing rice sheath blight disease. This is the first report on the blight disease of Azolla caused by R. solani in Korea.

Minority report; Diketopiperazines and Pyocyanin as Quorum Sensing Signals in Pseudomonas aeruginosa (Minority report; Pseudomonas aeruginosa의 정족수 인식(쿼럼 센싱) 신호물질로써의 Diketopiperazines과 Pyocyanin)

  • Lee, Joon-Hee
    • Korean Journal of Microbiology
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    • v.44 no.2
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    • pp.85-92
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    • 2008
  • Pseudomonas aeruginosa is an opportunistic human pathogen, causing a wide variety of infections including cystic fibrosis, microbial keratitis, and burn wound infections. The cell-to-cell signaling mechanism known as quorum sensing (QS) plays a key role in these infections and the QS systems of P. aeruginosa have been most intensively studied. While many literatures that introduce the QS systems of P. aeruginosa have mostly focused on two major acyl-homo serine lactone (acyl-HSL) QS signals, N-3-oxododecanoyl homoserine lactone (3OC12) and N-butanoyl homoserine lactone (C4), several new signal molecules have been discovered and suggested for their significant roles in signaling and virulence of P. aeruginosa. One of them is PQS (Pseudomonas quinolone signal; 2-heptyl-3-hydroxy-4-quinolone), which is now considered as a well-characterized major signal meolecule of P. aeruginosa. In addition, recent researches have also suggested some more putative signal molecules of P. aeruginosa, which are diketopiperazines (DKPs) and pyocyanin. DKPs are cyclic dipeptides and structurally diverse depending on what amino acids are involved in composition. Some DKPs from the culture supernatant of P. aeruginosa are suggested as new diffusible signal molecules, based on their ability to activate Vibrio fischeri LuxR biosensors that are previously considered specific for acyl-HSLs. Pyocyanin (1-hydroxy-5-methyl-phenazine), one of phenazine derivatives produced by P. aeruginosa is a characteristic blue-green pigment and redox-active compound. This has been recently suggested as a terminal signaling factor to upregulate some QS-controlled genes during stationary phase under the mediation of a transcription factor, SoxR. Here, details about these newly emerging signaling molecules of P. aeruginosa are discussed.

Examination of Antimicrobial Activity by Phaeobacter inhibens KJ-2 Isolated from a Marine Organism (해양 생물에서 분리된 Phaeobacter inhibens KJ-2의 항균 활성)

  • Kim, Yun-Beom;Kim, Dong-Hwi;Heo, Moon-Soo
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1161-1167
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    • 2017
  • In order to find a new antimicrobial bacterium, we performed screening for antimicrobial activity of bacteria isolated from the eggs of a sea hare. The newly identified strain was designated as Phaeobacter inhibens KJ-2, based on the biochemical characterization and 16S rRNA gene sequence analysis. A colony of P. inhibens KJ-2 showed a circular and ruler-like smooth form at the edge, and a brown color. However, when maintained with a longer incubation time, its coloring was transformed into dark brown. From the result of SEM, P. inhibens KJ-2 is a bacillus which has a length of $0.8{\sim}1.0{\mu}m$ and a width of $0.4{\sim}0.6{\mu}m$. The optimal growth and antimicrobial activity were observed by shaking the culture for 24 hr at $20^{\circ}C$, which showed potent activity against pathogenic bacteria including Vibrio logei, Vibrio campbellii, Vibrio mimicus, Vibrio vulnificus, and Vibrio salmonicida. The antimicrobial activity was proportional to the amount of produced acylated homoserine lactones (AHLs). Therefore, we suggest that production of antimicrobial materials from P. inhibens KJ-2 is regulated by Quorum sensing (QS).

Construction of an Oscillator Gene Circuit by Negative and Positive Feedbacks

  • Shen, Shihui;Ma, Yushu;Ren, Yuhong;Wei, Dongzhi
    • Journal of Microbiology and Biotechnology
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    • v.26 no.1
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    • pp.139-144
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    • 2016
  • Synthetic oscillators are gene circuits in which the protein expression will change over time. The delay of transcription, translation, and protein folding is used to form this kind of behavior. Here, we tried to design a synthetic oscillator by a negative feedback combined with a positive feedback. With the mutant promoter PLacC repressed by LacIq and PLux activated by AHL-bound LuxR, two gene circuits, Os-LAA and Os-ASV, were constructed and introduced into LacI-deleted E. coli DH5α cells. When glucose was used as the carbon source, a low level of fluorescence was detected in the culture, and the bacteria with Os-ASV showed no oscillation, whereas a small portion of those carrying Os-LAA demonstrated oscillation behavior with a period of about 68.3 ± 20 min. When glycerol was used as the carbon source, bacteria with Os-ASV demonstrated high fluorescence value and oscillation behavior with the period of about 121 ± 21 min.

Production of Acyl-Homoserine Lactone Quorum-Sensing Signals is Wide-Spread in Gram-Negative Methylobacterium

  • Poonguzhall, Poonguzhall;Selvaraj, Selvaraj;Madhaiyan, Munusamy;Sa, Tongmin
    • Journal of Microbiology and Biotechnology
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    • v.17 no.2
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    • pp.226-233
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    • 2007
  • Members of Methylobacterium, referred as pink-pigmented facultative methylotrophic bacteria, are frequently associated with terrestrial and aquatic plants, tending to form aggregates on the phyllosphere. We report here that the production of autoinducer molecules involved in the cell-to-cell signaling process, which is known as quorum sensing, is common among Methylobacterium species. Several strains of Methylobacterium were tested for their ability to produce N-acyl-homoserine lactone (AHL) signal molecules using different indicators. Most strains of Methylobacterium tested could elicit a positive response in Agrobacterium tumefaciens harboring lacZ fused to a gene that is regulated by autoinduction. The synthesis of these compounds was cell-density dependent, and the maximal activity was reached during the late exponential to stationary phases. The bacterial extracts were separated by thin-layer chromatography and bioassayed with A. tumefaciens NTI (traR, tra::lacZ749). They revealed the production of various patterns of the signal molecules, which are strain dependent. At least two signal molecules could be detected in most of the strains tested, and comparison of their relative mobilities suggested that they are homologs of N-octanoyl-$_{DL}$-homoserine lactone ($C_8-HSL$) and N-decanoyl-$_{DL}$-homoserine lactone ($C_{10}-HSL$).

Design of Quorum Quenching Microbial Vessel to Enhance Cell Viability for Biofouling Control in Membrane Bioreactor

  • Cheong, Won-Suk;Kim, Sang-Ryoung;Oh, Hyun-Suk;Lee, Sang H.;Yeon, Kyung-Min;Lee, Chung-Hak;Lee, Jung-Kee
    • Journal of Microbiology and Biotechnology
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    • v.24 no.1
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    • pp.97-105
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    • 2014
  • Quorum quenching (QQ) with a microbial vessel has recently been reported as an economically feasible biofouling control platform in a membrane bioreactor (MBR) for wastewater treatment. In this study, a quorum quenching MBR with a ceramic microbial vessel (CMV) was designed to overcome the extremely low F/M ratio inside a microbial vessel. The CMV was prepared with a monolithic ceramic microporous membrane and AHL-degrading QQ bacteria, Pseudomonas sp. 1A1. The "inner flow feeding mode" was introduced, under which fresh feed was supplied to the MBR only through the center lumen in the CMV. The inner flow feeding mode facilitated nutrient transport to QQ bacteria in the CMV and thus enabled relatively long-term maintenance of cell viability. The quorum quenching effect of the CMV on controlling membrane biofouling in the MBR was more pronounced with the inner flow feeding mode, which was identified by the slower increase in the transmembrane pressure as well as by the visual observation of a biocake that formed on the used membrane surface. In the QQ MBR with the CMV, the concentrations of extracellular polymeric substances were substantially decreased in the biocake on the membrane surface compared with those in the conventional MBR. The CMV also showed its potential with effective biofouling control over long-term operation of the QQ MBR.

Recovery of Poly(3-hydroxybutyrate) from Recombinant Escherichia coli by Autolysis with Bacteriophage Lambda (박테리오파아지의 세포용해작용을 이용한 재조합 대장균으로부터의 Poly(3-hydroxybutyrate) 회수)

  • 정옥희;한세광장용근이상엽
    • KSBB Journal
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    • v.10 no.5
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    • pp.533-539
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    • 1995
  • An autolytic system based on a thermally inducible phage lambda, λHL1, has been applied for the recovery of poly(3-hydroxybutyrate) [PHB] from a recombinant Escherichia coli XL1-Blue, harbouring a plasmid (pSYL105) containing the Alcaligenes eutrophus PHB biosynthesis genes. The lytic capability ofλHL1 was evaluated in flask culture for both lysogens, XL1-Blue (λHL1) and XL1-Blue (λHL1, pSYL105). When the optical density of culture at 600nm(OD600) reached 0.2, cell lysis was induced by increasing the temperature from $30^{\circ}C$ to $42^{\circ}C$. Most cells of XL1-Blue ($\lambda$HL1) were lysed by the autolytic system in an hour after the thermal induction, while the lytic efficiency was slightly lower for XLl-Blue (λHL1, pSYL105). The existence of pSYL105 in cells seemed to inhibit, to some extent, the lytic capability of λHL1 even at low PHB content. The lylic efficiency remarkably decreased as the induction was delayed to allow PHB accumulation. When a chemical induction using 2% (v/v) chloroform was introduced after an hours of thermal induction, we could obtain a good lytic efficiency.

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Detection of a Quorum-Sensing Inhibitor from the Natural Products (천연물로부터 Quorum Sensing 저해제의 탐색)

  • Kim, Tae-Woo;Cha, Ji-Young;Lee, Jun-Seung;Min, Bok-Kee;Baik, Hyung-Suk
    • Journal of Life Science
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    • v.18 no.2
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    • pp.206-212
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    • 2008
  • The quorum sensing (QS) regulatory network has been the subject of extensive studies during recent years and has also attracted a lot of attention because it both positively and negatively regulates various putative virulence factors, although initially considered to be a specialized system of Vibrio fischeri and related species. In this study, to identify the novel materials which inhibit QS system of microorganisms, extracts of eighteen natural products were tested by bioassay using N-(3-oxohexanoyl)-$_L$-homoserine lactone and N-(3-oxooctanoyl)-$_L$-homoserine lactone synthesized in this experiment and an Agrobacterium tumefaciens NT1 biosensor strain containing a traI::lacZ fusion. The result indicated that the extracts of cabbage, leek, and onion exhibited the QS inhibition activity. Thus, materials contained in the extracts were isolated via recycling preparative HPLC and were purified via a JAIGEL-LS255 column. The common fraction corresponding to a peak of the 83 min point of them quenched the quorum sensing of A. tumefaciens NT1 biosensor strain in ABMM containing X-gal and was designated quorum sensing inhibitor-83 min (QSI-83). The QSI-83 exhibited the heat stability and did not inhibit the growth of A. tumefaciens NTl. Furthermore, thin layer chromatography (TLC) results suggested that these novel materials may be antagonists of N-acyl homoserine lactone or may inhibit the QS autoinducer synthesis by Pseudomonas syringae pv. tabaci.

Potential of fascaplysin and palauolide from Fascaplysinopsis cf reticulata to reduce the risk of bacterial infection in fish farming

  • Mai, Tepoerau;Toullec, Jordan;Wynsberge, Simon Van;Besson, Marc;Soulet, Stephanie;Petek, Sylvain;Aliotti, Emmanuelle;Ekins, Merrick;Hall, Kathryn;Erpenbeck, Dirk;Lecchini, David;Beniddir, Mehdi A.;Saulnier, Denis;Debitus, Cecile
    • Fisheries and Aquatic Sciences
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    • v.22 no.12
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    • pp.30.1-30.11
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    • 2019
  • Marine natural products isolated from the sponge Fascaplysinopsis cf reticulata, in French Polynesia, were investigated as an alternative to antibiotics to control pathogens in aquaculture. The overuse of antibiotics in aquaculture is largely considered to be an environmental pollution, because it supports the transfer of antibiotic resistance genes within the aquatic environment. One environmentally friendly alternative to antibiotics is the use of quorum sensing inhibitors (QSIs). Quorum sensing (QS) is a regulatory mechanism in bacteria which control virulence factors through the secretion of autoinducers (AIs), such as acyl-homoserine lactone (AHL) in gram-negative bacteria. Vibrio harveyi QS is controlled through three parallel pathways: HAI-1, AI-2, and CAI-1. Bioassay-guided purification of F. cf reticulata extract was conducted on two bacterial species, i.e., Tenacibaculum maritimum and V. harveyi for antibiotic and QS inhibition bioactivities. Toxicity bioassay of fractions was also evaluated on the freshwater fish Poecilia reticulata and the marine fish Acanthurus triostegus. Cyclohexanic and dichloromethane fractions of F. cf reticulata exhibited QS inhibition on V. harveyi and antibiotic bioactivities on V. harveyi and T. maritimum, respectively. Palauolide (1) and fascaplysin (2) were purified as major molecules from the cyclohexanic and dichloromethane fractions, respectively. Palauolide inhibited QS of V. harveyi through HAI-1 QS pathway at 50 ㎍ ml-1 (26 μM), while fascaplysin affected the bacterial growth of V. harveyi (50 ㎍ ml-1) and T. maritimum (0.25 ㎍). The toxicity of fascaplysin-enriched fraction (FEF) was evaluated and exhibited a toxic effect against fish at 50 ㎍ ml-1. This study demonstrated for the first time the QSI potential of palauolide (1). Future research may assess the toxicity of both the cyclohexanic fraction of the sponge and palauolide (1) on fish, to confirm their potential as alternative to antibiotics in fish farming.