• Title/Summary/Keyword: GacS

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The Sensor Kinase GacS Negatively Regulates Flagellar Formation and Motility in a Biocontrol Bacterium, Pseudomonas chlororaphis O6

  • Kim, Ji Soo;Kim, Yong Hwan;Anderson, Anne J.;Kim, Young Cheol
    • The Plant Pathology Journal
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    • v.30 no.2
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    • pp.215-219
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    • 2014
  • The GacS/GacA two component system regulates various traits related to the biocontrol potential of plant-associated pseudomonads. The role of the sensor kinase, GacS, differs between strains in regulation of motility. In this study, we determined how a gacS mutation changed cell morphology and motility in Pseudomonas chlororaphis O6. The gacS mutant cells were elongated in stationary-phase compared to the wild type and the complemented gacS mutant, but cells did not differ in length in logarithmic phase. The gacS mutant had a two-fold increase in the number of flagella compared with the wild type strain; flagella number was restored to that of the wild type in the complemented gacS mutant. The more highly flagellated gacS mutant cells had greater swimming motilities than that of the wild type strain. Enhanced flagella formation in the gacS mutant correlated with increased expression of three genes, fleQ, fliQ and flhF, involved in flagellar formation. Expression of these genes in the complemented gacS mutant was similar to that of the wild type. These findings show that this root-colonizing pseudomonad adjusts flagella formation and cell morphology in stationary-phase using GacS as a major regulator.

Effect of gacS and gacA Mutations on Colony Architecture, Surface Motility, Biofilm Formation and Chemical Toxicity in Pseudomonas sp. KL28

  • Choi, Kyung-Soon;Veeraragouda, Yaligara;Cho, Kyoung-Mi;Lee, Soo-O;Jo, Geuk-Rae;Cho, Kyung-Yun;Lee, Kyoung
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.492-498
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    • 2007
  • GacS and GacA proteins form a two component signal transduction system in bacteria. Here, Tn5 transposon gacS and gacA (Gac) mutants of Pseudomonas sp. KL28, an alkylphenol degrader, were isolated by selecting for smooth colonies of strain KL28. The mutants exhibited reduced ability to migrate on a solid surface. This surface motility does not require the action of flagella unlike the well-studied swarming motility of other Pseudomonas sp. The Gac mutants also showed reduced levels of biofilm and pellicle formation in liquid culture. In addition, compared to the wild type KL28 strain, these mutants were more resistant to high concentrations of m-cresol but were more sensitive to $H_2O_2$, which are characteristics that they share with an rpoS mutant. These results indicate that the Gac regulatory cascade in strain KL28 positively controls wrinkling morphology, biofilm formation, surface translocation and $H_2O_2$ resistance, which are important traits for its capacity to survive in particular niches.

Proteomic Analysis of the GacA Response Regulator in Pseudomonas chlororaphis O6

  • Anderson, Anne J.;Kim, Young Cheol
    • Research in Plant Disease
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    • v.24 no.2
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    • pp.162-169
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    • 2018
  • The GacS/GacA system in the root colonizer Pseudomonas chlororaphis O6 is a key regulatory system of many traits relevant to the plant probiotic nature of this bacterium. The work in this paper elucidates proteins using proteomics approach in P. chlororaphis O6 under the control of the cytoplasmic regulatory protein, GacA. A gacA mutant of P. chlororaphis O6 showed loss in production of phenazines, acyl homoserine lactones, hydrogen cyanide, and protease, changes that were associated with reduced in vitro antifungal activity against plant fungal pathogens. Production of iron-chelating siderophore was significantly enhanced in the gacA mutant, also paralleling changes in a gacS mutant. However, proteomic analysis revealed proteins (13 downregulated and 7 upregulated proteins in the mutant compared to parental strain) under GacA control that were not apparent by a proteomic study of a gacS mutant. The putative identity of the downregulated proteins suggested that a gacA mutant would have altered transport potentials. Notable would be a predicted loss of type-VI secretion and PEP-dependent transport. Study of mutants of these GacA-regulated proteins will indicate further the features required for probiotic potential in this rhizobacterium.

Utilization of Element-doping Titania-impregnated Granular Activated Carbon in a Plug-flow System for Removal of BTEX

  • Jo, Wan-Kuen;Shin, Seung-Ho;Hwang, Eun-Song;Yang, Sung-Bong
    • Asian Journal of Atmospheric Environment
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    • v.4 no.3
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    • pp.177-188
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    • 2010
  • The use of an activated carbon (AC) system alone has the limitation that the pollutants are not eliminated but only transferred to another phase with the consumed AC becoming hazardous waste itself. Therefore, the present study investigated the feasibility of using a combined system of granular AC (GAC) with S-doped visible-light-induced $TiO_2$ (GAC/S-doped $TiO_2$) to clean monocyclic aromatic hydrocarbons (MAHs) with concentrations at $\leq$ 3 mg $m^{-3}$, using a continuous air-flow reactor. This study conducted three different experiments: an adsorption test of pure GAC and GAC/S-doped $TiO_2$; a long-term adsorptional photocatalytic (AP) activity test of GAC/S-doped $TiO_2$; and an AP activity test of GAC/S-doped $TiO_2$ under different conditions. For the AP activity test, three parameters were evaluated: various weights of GAC/S-doped $TiO_2$ (0.9, 4.4, and 8.9 g); various flow rates (FRs) (0.5, 1 and 2 L $min^{-1}$); and various input concentrations (ICs) of the target MAHs (0.1, 1, 2 and 3 mg $m^{-3}$). The adsorption efficiencies were similar for the pure GAC and GAC/S-doped $TiO_2$ reactors, suggesting that S-doped $TiO_2$ particles on GAC surfaces do not significantly interfere with the adsorption capacity of GAC. Benzene exhibited a clear AP activity, whereas no other target MAHs did. In most cases, the AP efficiencies for the target MAHs did not significantly vary with an increase in weight, thereby suggesting that, under the weight range tested in this study, the weights or FRs are not important parameters for AP efficiency. However, ICs did influence the AP efficiencies.

Analysis of Enterococcus faecalis Attachment to Granular Activated Carbon with a Column Experiment (칼럼실험에 의한 입상활성탄에서 Enterococcus faecalis의 부착 연구)

  • Kim, Hyon-Chong;Park, Seong-Jik;Lee, Chang-Gu;Han, Yong-Un;Kim, Song-Bae
    • Journal of Korean Society of Environmental Engineers
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    • v.31 no.2
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    • pp.119-124
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    • 2009
  • The aim of this study was to investigate the influence of ionic strength and iron impregnation on the attachment of Enterococcus faecalis to granular activated carbon (GAC). Column experiments were performed to examine bacterial adhesion to coconutbased GAC (c-GAC), iron-impregnated c-GAC (fc-GAC), acid-washed c-GAC (a-GAC) and iron-impregnated a-GAC (fa-GAC) under two different solution (NaCl 1, 10 mM) conditions. Results showed that bacterial mass recovery in c-GAC decreased from 77.3 to 61.6% while in a-GAC it decreased from 71.6 to 32.3% with increasing ionic strength from 1 to 10 mM. This indicates that bacterial attachment to GAC can be enhanced with increasing ionic strength. Results also showed that the mass recoveries in fc-GAC were 62.6% (1 mM) and 53.3% (10 mM) while they were 50.8% (1 mM) and 16.9%(10 mM) in fa-GAC, which were lower than those in c-GAC and a-GAC. This demonstrates that bacterial adhesion to GAC can be enhanced through iron impregnation. This study provides information regarding the effects of ionic strength and iron impregnation on bacterial attachment to GAC. Furthermore, this study will advance our knowledge of bacterial removal in surface-modified granular media.

Proteomic Analysis of a Global Regulator GacS Sensor Kinase in the Rhizobacterium, Pseudomonas chlororaphis O6

  • Kim, Chul Hong;Kim, Yong Hwan;Anderson, Anne J.;Kim, Young Cheol
    • The Plant Pathology Journal
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    • v.30 no.2
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    • pp.220-227
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    • 2014
  • The GacS/GacA system in the root colonizer Pseudomonas chlororaphis O6 is a key regulator of many traits relevant to the biocontrol function of this bacterium. Proteomic analysis revealed 12 proteins were down-regulated in a gacS mutant of P. chlororaphis O6. These GacS-regulated proteins functioned in combating oxidative stress, cell signaling, biosynthesis of secondary metabolism, and secretion. The extent of regulation was shown by real-time RT-PCR to vary between the genes. Mutants of P. chlororaphis O6 were generated in two GacS-regulated genes, trpE, encoding a protein involved in tryptophan synthesis, and prnA, required for conversion of tryptophan to the antimicrobial compound, pyrrolitrin. Failure of the trpE mutant to induce systemic resistance in tobacco against a foliar pathogen causing soft rot, Pectobacterium carotovorum SCCI, correlated with reduced colonization of root surfaces implying an inadequate supply of tryptophan to support growth. Although colonization was not affected by mutation in the prnA gene, induction of systemic resistance was reduced, suggesting that pyrrolnitrin was an activator of plant resistance as well as an antifungal agent. Study of mutants in the other GacS-regulated proteins will indicate further the features required for biocontrol-activity in this rhizobacterium.

A Study of the Regeneration of Spent GAC using an Electrochemical Method (전기화학적 방법을 이용한 Spent Granular Activated Carbon (GAC)의 재생 연구)

  • Lee, Sangmin;Joo, Soobin;Jo, Youngsoo;Oh, Yeji;Kim, Hyungjun;Shim, Intae
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.42 no.4
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    • pp.481-491
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    • 2022
  • This study investigates the characteristics of the GAC adsorption behavior during the operation of a multi-stage cross-flow filtration and GAC adsorption process for the purpose of devising an advanced treatment of combined sewer overflows (CSOs) and evaluates the regeneration efficiency of spent GAC that has reached the design breakpoint. During the filtration process, suspended substances are easily removed, but dissolved organic substances are not removed, necessitating a process capable of removing dissolved organic substances for the advanced treatment of CSOs. In general, GAC adsorption has been applied under low-concentration organic conditions, such as for water purification and tertiary treatments of sewage, and has rarely been applied under conditions with high organic concentrations, such as with sewage or CSOs. Accordingly, this study will provide a new and interesting experience. Also in this study, the continuous operation and breakthrough characteristics of GAC according to the strength of the inflow organic matter were investigated, electrochemical regeneration was applied to the used GAC, and the regeneration efficiency was evaluated through desorption and re-adsorption tests. The results showed that the breakthrough period was 21 days under high concentration conditions, 28 days at medium concentrations, and 32 days under low concentration conditions. The desorption of adsorbed organic matter through electrolysis occurred in the range of 188 to 609 mgCOD/L depending on the electrolysis conditions, and the effect of the electrolyte type led to the finding that NaOH was slightly higher than H2O2.

The global regulator GacS of a biological bacterium Pseudomonas chlororaphis O6 regulates expression of the stationary-phase sigma factor rpoS and reduces survival in oxidative stress.

  • Kang, Beom-Ryong;Cho, Baik-Ho;Kim, Young-Cheol
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.100.2-101
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    • 2003
  • The global regulator, GacS (global antibiotic and cyanide sensor kinase), was required for the increased resistance to hydrogen peroxide occurring as cultures of the rhizobacterium, P. chlororaphis O6, matured. Specific stationary-phase peroxidase and catalase isozymes were absent in the GacS mutant, whereas a manganese-superoxide dismutase isozyme was expressed earlier and to a great extent than wild type. In the wild type cell, transcript accumulation of rpoS was higher in late logarithmic-phase cells than cells from mid logarithmic- or stationary-phase. Transcripts from rpoS in the GacS mutant were reduced in each of these growth phases compared to the wild type expression. The down stream sequence from rpoS lacked sequences encoding a small RNA, rsmZ, found in other pseudomonads and implicated in control of genes activated by the GacS system. These findings suggest that GacS-mediated regulation of RpoS plays role in control of oxidative stress in P. chlororaphis O6 by as yet an unknown mechanism.

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A two-component sensor kinase (GacS) mediated signal transduction pathway involved in production of antifungal compounds in Pseudomonas chlororaphis O6.

  • Kang, Beom-Ryong;Lee, Jung-Hoon;Kim, Hyun-Jung;Cho, Baik-Ho;Kim, Young-Cheol
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.99.1-100
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    • 2003
  • E. intermedium Blocontrol activity of a P. chlororaphis rhizobacteium O6, depends to the synthesis of extracellular secondary metabolites and exoenzymes, thought to antagonize the pathogenicity of a variety of phytopathogenic fungi. The production of secondary metabolites and exoenzymes in O6, depends essentially on the GacS-mediated signal transduction pathway, which activates largely unknown signal transduction pathway. To exploit the GacS-mediated signal transdcution pathway involved in activation of ph genes that are necessary for biosynthesis of phenazine from P. chlororaphis O6, we cloned and sequenced the phz operon, rpoS gene encoding stationary specific sigma factor, ppx gene encoding polyphosphatase, and lon gene encoding ion protease. Expression of each gene in wild type and GacS mutant were analyzed by RT-PCR. Transcripts from rpoS, phzI enconing acylhomoserine lactone (AHL) synthase, and ph structural genes in the GacS mutant were reduced in each of these growth phases compared to the wild type. The GacS or Lon mutant was found to be deficient in the production of phenzines, exoenzymes, and the acylhomoserine lactone. These mutants were not complemented by ph operon and addition of exogenous AHL. These results indicate that the GacS global regulatory systems controls phenazine production at multiple levels. Future research will focus to identifying the GacS-mediated regulatory cascade involving in production of phenazine in P. chlororaphis.

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Effect of Ozonation on Removal of Dissolved Organic Matter by Granular Activated Carbon Process (오존공정이 입상활성탄공정에서 용존유기물질의 제거에 미치는 영향)

  • Ahn, Hyo-Won;Chae, Seon-Ha;Wang, Chang-Keun;Lim, Jae-Lim
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.6
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    • pp.601-608
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    • 2008
  • The objective of this study was to evaluate the effect of ozonation as pretreatment on the removal of dissolved or biodegradable organic matter(DOM or BOM), the variance of DOM fractionation, and microbial regrowth by pilot-scale granular activated carbon processes in which adsorption and biodegradability was proceeding due to long time operation. Regardless of point of ozonation applied, GAC processes with ozonation(i.e., Ozonation combined with GAC Filter-adsorber; Pre O$_3$ + F/A, Ozonation combined with GAC adsorber; Post O$_3$ + GAC) compared with GAC processes without ozonation(i.e., GAC Filter-adsorber; F/A, GAC adsorber; GAC) removed approximately 10 to 20% more of DOC, hydrophilic DOM(HPI), BDOC and AOC after long period of operation that biological activity was assumed to happen. Ozonation was not found to have a significant effect on the removal of DOC, but caused the decrease of AOC by approximately 20%. It was found that the fixed bacterial biomass on GAC media did not show a significant difference between the GAC with ozonation and GAC without ozonation as pre-treatment, whereas the HPC of column effluent was more biostable at Post O$_3$ + GAC compared with F/A or GAC.