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Involvement of quorum sensing genes in biofilm development and degradation of polycyclic aromatic hydrocarbons by a marine bacterium Pseudomonas aeruginosa N6P6

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Abstract

Biofilm-forming and acyl homoserine lactone (AHL) synthase-positive Pseudomonas aeruginosa N6P6 was isolated from seawater after selective enrichment with two polycyclic aromatic hydrocarbons (PAHs), viz. phenanthrene and pyrene. AHL synthesis was detected qualitatively using bioreporter strains. This marine bacterium putatively synthesized N-(3-oxododecanoyl)-l-homoserine lactone and N-butyryl-l-homoserine lactone, which were identified by TLC, GC-MS, and HPLC. Two quorum sensing (QS) genes coding for AHL synthase, i.e., lasI and rhlI, were identified in the bacterium. lasI and rhlI gene expression was studied during biofilm mode of growth at different phases using quantitative real-time PCR (qRT-PCR). The expression of lasI increased with increase in biofilm growth. In contrast, the expression of rhlI decreased during log phase of biofilm growth. The changes in lasI/rhlI expression level had significant effects (P < 0.05) on biofilm architecture and subsequent PAH degradation rate. Degradation of phenanthrene and pyrene by P. aeruginosa N6P6 was affected by biofilm growth and lasI expression. The respective phenanthrene degradation for 15, 24, 48, and 72 h old biofilm after 7 days was 21.5, 54.2, 85.6, and 85.7 %. However, the corresponding pyrene degradation was 15, 18.28, 47.56, and 46.48 %, respectively, after 7 days. A significant positive correlation (P < 0.05) was observed between lasI expression and PAHs degradation. However, in the presence of tannic acid, a QS inhibitor (QSI), PAHs degradation, biofilm formation, and pyocyanin production reduced significantly which confirmed the pivotal role of QS in biodegradation of PAHs. The findings suggest that AHLs play a pivotal role during biofilm development and subsequent bioremediation of PAHs.

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References

  • Akbar S, Sultan S, Kertesz M (2014) Bacterial community analysis in chlorpyrifos enrichment cultures via DGGE and use of bacterial consortium for CP biodegradation. World J Microb Biot 30:2755–2766

    Article  CAS  Google Scholar 

  • Biswa P, Doble M (2013) Production of acylated homoserine lactone by gram-positive bacteria isolated from marine water. FEMS Microbiol Lett 343:34–41

    Article  CAS  PubMed  Google Scholar 

  • Cady NC, McKean KA, Behnke J, Kubec R, Mosier AP, Kasper SH, Burz DS, Musah RA (2012) Inhibition of biofilm formation, quorum sensing and infection in Pseudomonas aeruginosa by natural products-inspired organosulfur compounds. PLoS One 7:8

    Article  Google Scholar 

  • Cataldi TR, Bianco G, Frommberger M, Schmitt-Kopplin P (2004) Direct analysis of selected N-acyl-L-homoserine lactones by gas chromatography/mass spectrometry. Rapid Commun Mass Spectrum 18:1341–1344

    Article  CAS  Google Scholar 

  • Chakraborty J, Das S (2014) Characterization and cadmium-resistant gene expression of biofilm-forming marine bacterium Pseudomonas aeruginosa JP-11. Environ Sci Pollut Res Int 21:14188–141201

    Article  CAS  PubMed  Google Scholar 

  • Chang CY, Krishnan T, Wang H, Chen Y, Yin WF, Chong YM, Chan KG (2014) Non-antibiotic quorum sensing inhibitors acting against N-acyl homoserine lactone synthase as druggable target. Scientific Reports, 4.

  • Chong YM, Yin WF, Ho CY, Mustafa MR, Hadi AHA, Awang K, Chan KG (2011) Malabaricone C from Myristica cinnamomea exhibits anti-quorum sensing activity. J Nat Prod 74(10):2261–2264

    Article  CAS  PubMed  Google Scholar 

  • Chong TM, Koh CL, Sam CK, Choo YM, Yin WF, Chan KG (2012) Characterization of quorum sensing and quorum quenching soil bacteria isolated from Malaysian tropical montane forest. Sensors 12(4):4846–4859

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Christensen GD, Simpson WA, Bisno AL, Beachey EH (1982) Adherence of slime-producing strains of Staphylococcus epidermidis to smooth surfaces. Infect Immun 37(1):318–326

  • Das S (2014) Microbial Biodegradation and Bioremediation. 1st Edition: Elsevier, Germany

  • Das K, Mukherjee AK (2007) Differential utilization of pyrene as the sole source of carbon by Bacillus subtilis and Pseudomonas aeruginosa strains: role of biosurfactants in enhancing bioavailability. J Appl Microbiol 102:195–203

    Article  CAS  PubMed  Google Scholar 

  • Das T, Manefield M (2012) Pyocyanin promotes extracellular DNA release in Pseudomonas aeruginosa. PLoS ONE 7(10)

  • Dasgupta D, Ghosh R, Sengupta TK (2013) Biofilm-mediated enhanced crude oil degradation by newly isolated Pseudomonas species. ISRN Biotechnol 2013: Article ID 250749

  • Dash HR, Das S (2014) Bioremediation potential of mercury by Bacillus species isolated from marine environment and wastes of steel industry. Biorem J 18:204–212

    Article  CAS  Google Scholar 

  • Dash HR, Mangwani N, Chakraborty J, Kumari S, Das S (2013) Marine bacteria: potential candidates for enhanced bioremediation. Appl Microbiol Biotechnol 97(2):561–571

    Article  CAS  PubMed  Google Scholar 

  • Davies DG, Parsek MR, Pearson JP, Iglewski BH, Costerton JW, Greenberg EP (1998) The involvement of cell-to-cell signals in the development of a bacterial biofilm. Science 280:295–298

    Article  CAS  PubMed  Google Scholar 

  • De Kievit T (2009) Quorum sensing in Pseudomonas aeruginosa biofilms. Environ Microbiol 11:279–288

    Article  PubMed  Google Scholar 

  • De Kievit TR, Gillis R, Marx S, Brown C, Iglewski BH (2001) Quorum-sensing genes in Pseudomonas aeruginosa biofilms: their role and expression patterns. Appl Environ Microbiol 67:1865–1873

    Article  PubMed Central  PubMed  Google Scholar 

  • Dong YH, Wang LH, Xu JL, Zhang HB, Zhang XF, Zhang LH (2001) Quenching quorum-sensing-dependent bacterial infection by an N-acyl homoserine lactonase. Nature 411:813–817

    Article  CAS  PubMed  Google Scholar 

  • Dusane DH, Zinjarde SS, Venugopalan VP, McLean RJ, Weber MM, Rahman PK (2010) Quorum sensing: implications on rhamnolipid biosurfactant production. Biotechnol Genet Eng Rev 27:159–184

    Article  CAS  PubMed  Google Scholar 

  • Feng L, Wu Z, Yu X (2013) Quorum sensing in water and wastewater treatment biofilms. J Environ Biol 34:437–444

    CAS  PubMed  Google Scholar 

  • Gao J, Ma A, Zhuang X, Zhuang G (2014) An N-acyl homoserine lactone synthase in the ammonia-oxidizing bacterium Nitrosospira multiformis. Appl Environ Microbiol 80:951–958

    Article  PubMed Central  PubMed  Google Scholar 

  • Gupta R, Schuster M (2012) Quorum sensing modulates colony morphology through alkyl quinolones in Pseudomonas aeruginosa. BMC Microbiol 12:1471–2180

    Article  Google Scholar 

  • Guzzella L, Roscioli C, Vigano L, Saha M, Sarkar SK, Bhattacharya A (2005) Evaluation of the concentration of HCH, DDT, HCB, PCB and PAH in the sediments along the lower stretch of Hugli estuary, West Bengal, northeast India. Environ Int 31:523–534

    Article  CAS  PubMed  Google Scholar 

  • Hogardt M, Roeder M, Schreff AM, Eberl L, Heesemann J (2004) Expression of Pseudomonas aeruginosa exoS is controlled by quorum sensing and RpoS. Microbiology 150:843–851

    Article  CAS  PubMed  Google Scholar 

  • Huang YL, Ki JS, Lee OO, Qian PY (2009) Evidence for the dynamics of acyl homoserine lactone and AHL-producing bacteria during subtidal biofilm formation. ISME J 3:296–304

    Article  CAS  PubMed  Google Scholar 

  • Huang Y, Zeng Y, Yu Z, Zhang J, Feng H, Lin X (2013) In silico and experimental methods revealed highly diverse bacteria with quorum sensing and aromatics biodegradation systems—a potential broad application on bioremediation. Bioresour Technol 148:311–316

    Article  CAS  PubMed  Google Scholar 

  • IARC Monographs on the evaluation of the carcinogenic risk of chemicals to humans (1983) Polynuclear aromatic compounds, Part 1, Chemical, environmental and experimental data 32:1-453

  • Jain K, Parida S, Mangwani N, Dash H, Das S (2013) Isolation and characterization of biofilm-forming bacteria and associated extracellular polymeric substances from oral cavity. Ann Microbiol 63:1553–1562

    Article  CAS  Google Scholar 

  • Jones SM, Dang TT, Martinuzzi R (2009) Use of quorum sensing antagonists to deter the formation of crystalline Proteus mirabilis biofilms. Int J Antimicrob Ag 34(4):360–364

  • Kang YS, Park W (2010) Contribution of quorum-sensing system to hexadecane degradation and biofilm formation in Acinetobacter sp. strain DR1. J Appl Microbiol 109:1650–1659

    CAS  PubMed  Google Scholar 

  • Karamalidis AK, Evangelou AC, Karabika E, Koukkou AI, Drainas C, Voudrias EA (2010) Laboratory scale bioremediation of petroleum-contaminated soil by indigenous microorganisms and added Pseudomonas aeruginosa strain spet. Bioresour Technol 101:6545–6552

    Article  CAS  PubMed  Google Scholar 

  • Li Y, Qu HP, Liu JL, Wan HY (2014) Correlation between group behavior and quorum sensing in Pseudomonas aeruginosa isolated from patients with hospital-acquired pneumonia. J Thorac Dis 6:810–817

    PubMed Central  PubMed  Google Scholar 

  • Liao NQ, Li HM (2013) Conceivable bioremediation techniques based on quorum sensing. Appl Mech Mater 295:39–44

    Article  Google Scholar 

  • Ma Y, Wang L, Shao Z (2006) Pseudomonas, the dominant polycyclic aromatic hydrocarbon degrading bacteria isolated from Antarctic soils and the role of large plasmids in horizontal gene transfer. Environ Microbiol 8:455–465

    Article  CAS  PubMed  Google Scholar 

  • Mangwani N, Dash HR, Chauhan A, Das S (2012) Bacterial quorum sensing: functional features and potential applications in biotechnology. J Mol Microbiol Biotechnol 22:215–227

    Article  CAS  PubMed  Google Scholar 

  • Mangwani N, Shukla SK, Rao TS, Das S (2014a) Calcium-mediated modulation of Pseudomonas mendocina NR802 biofilm influences the phenanthrene degradation. Colloids Surf B Biointerfaces 114:301–309

    Article  CAS  PubMed  Google Scholar 

  • Mangwani N, Shukla SK, Kumari S, Rao TS, Das S (2014b) Characterization of Stenotrophomonas acidaminiphila NCW-702 biofilm for implication in the degradation of polycyclic aromatic hydrocarbons. J Appl Microbiol 18:12602

    Google Scholar 

  • McClean KH, Winson MK, Fish L, Taylor A, Chhabra SR, Camara M, Daykin M, Lamb JH, Swift S, Bycroft BW, Stewart GS, Williams P (1997) Quorum sensing and Chromobacterium violaceum: exploitation of violacein production and inhibition for the detection of N-acylhomoserine lactones. Microbiology 143:3703–3711

    Article  CAS  PubMed  Google Scholar 

  • Morici LA, Carterson AJ, Wagner VE, Frisk A, Schurr JR, Honer zu Bentrup K, Hassett DJ, Iglewski BH, Sauer K, Schurr MJ (2007) Pseudomonas aeruginosa AlgR represses the Rhl quorum-sensing system in a biofilm-specific manner. J Bacteriol 189:7752–7764

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Nasser W, Reverchon S (2007) New insights into the regulatory mechanisms of the LuxR family of quorum sensing regulators. Anal Bioanal Chem 387:381–390

    Article  CAS  PubMed  Google Scholar 

  • Nawaz MS, Chapatwala KD (1991) Simultaneous degradation of acetonitrile and biphenyl by Pseudomonas aeruginosa. Can J Microbiol 37:411–418

    Article  CAS  PubMed  Google Scholar 

  • Nomura M, Gourse R, Baughman G (1984) Regulation of the synthesis of ribosomes and ribosomal components. Annu Rev Biochem 53:75–117

    Article  CAS  PubMed  Google Scholar 

  • Paliwal V, Puranik S, Purohit HJ (2012) Integrated perspective for effective bioremediation. Appl Biochem Biotechnol 166:903–924

    Article  CAS  PubMed  Google Scholar 

  • Pan X, Liu J, Zhang D (2010) Binding of phenanthrene to extracellular polymeric substances (EPS) from aerobic activated sludge: a fluorescence study. Colloids Surf B: Biointerfaces 80:103–106

    Article  CAS  PubMed  Google Scholar 

  • Paraneeiswaran A, Shukla SK, Subba Rao T, Prashanth K (2014) Removal of toxic Co-EDTA complex by a halophilic solar-salt-pan isolate Pseudomonas aeruginosa SPB-1. Chemosphere 95:503–510

    Article  CAS  PubMed  Google Scholar 

  • Pavlović Z, Muck T, Hladnik A, Karlović I (2012) A comparative study of offset plate quality parameters using image processing and analytical methods. Acta Polytech Hung 9:181–193

    Google Scholar 

  • Pérez-Osorio AC, Williamson KS, Franklin MJ (2010) Heterogeneous rpoS and rhlR mRNA levels and 16S rRNA/rDNA (rRNA gene) ratios within Pseudomonas aeruginosa biofilms, sampled by laser capture microdissection. J Bacteriol 192:2991–3000

    Article  PubMed Central  PubMed  Google Scholar 

  • Reis RS, Pereira AG, Neves BC, Freire DMG (2011) Gene regulation of rhamnolipid production in Pseudomonas aeruginosa—a review. Bioresour Technol 102:6377–6384

    Article  CAS  PubMed  Google Scholar 

  • Ron EZ, Rosenberg E (2002) Biosurfactants and oil bioremediation. Curr Opin Biotechnol 13:249–252

    Article  CAS  PubMed  Google Scholar 

  • Roostan Z, Safahieh A, Mojodi F, Zolgharnein H, Ghanemi K, Abiar H (2012) Phenanthrene biodegradation by Pseudomonas aeruginosa and Bacillus subtilis isolated from Persian gulf sediments. Afr J Microbiol Res 6:4585–4591

    CAS  Google Scholar 

  • Sakuragi Y, Kolter R (2007) Quorum-sensing regulation of the biofilm matrix genes (pel) of Pseudomonas aeruginosa. J Bacteriol 189:5383–5386

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Sarabhai S, Sharma P, Capalash N (2013) Ellagic acid derivatives from Terminalia chebula Retz. Downregulate the expression of quorum sensing genes to attenuate Pseudomonas aeruginosa PAO1 virulence. PLoS One 8(1):e53441

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Shaw PD, Ping G, Daly SL, Cha C, Cronan Jr JE, Rinehart KL, Farrand SK (1997) Detecting and characterizing N-acyl-homoserine lactone signal molecules by thin-layer chromatography. Proc Natl Acad Sci U S A 94:6036–6041

  • Shimada K, Itoh Y, Washio K, Morikawa M (2012) Efficacy of forming biofilms by naphthalene degrading Pseudomonas stutzeri T102 toward bioremediation technology and its molecular mechanisms. Chemosphere 87:226–233

    Article  CAS  PubMed  Google Scholar 

  • Shu C, Sun S, Chen J, Chen J, Zhou E (2014) Comparison of different methods for total RNA extraction from sclerotia of Rhizoctonia solani. Electron J Biotechnol 17:50–54

    Article  Google Scholar 

  • Shukla SK, Rao TS (2013) Dispersal of Bap-mediated Staphylococcus aureus biofilm by proteinase K. J Antibiot 66(2):55–60

    Article  Google Scholar 

  • Shukla SK, Mangwani N, Rao TS, Das S (2014) Biofilm-mediated bioremediation of polycyclic aromatic hydrocarbons. In: Das S (ed), Microbial Biodegradation and Bioremediation, 1st edn. Elsevier, pp 203–232

  • Singh R, Paul D, Jain RK (2006) Biofilms: implications in bioremediation. Trends Microbiol 14:389–397

    Article  CAS  PubMed  Google Scholar 

  • Suárez-Moreno ZR, Devescovi G, Myers M, Hallack L, Mendonça-Previato L, Caballero-Mellado J, Venturi V (2010) Commonalities and differences in regulation of N-acyl homoserine lactone quorum sensing in the beneficial plant-associated Burkholderia species cluster. Appl Environ Microbiol 76:4302–4317

    Article  PubMed Central  PubMed  Google Scholar 

  • Venturi V (2006) Regulation of quorum sensing in Pseudomonas. FEMS Microbiol Rev 30:274–291

    Article  CAS  PubMed  Google Scholar 

  • Wagner VE, Bushnell D, Passador L, Brooks AI, Iglewski BH (2003) Microarray analysis of Pseudomonas aeruginosa quorum-sensing regulons: effects of growth phase and environment. J Bacteriol 185:2080–2095

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Watson WT, Minogue TD, Val DL, von Bodman SB, Churchill ME (2002) Structural basis and specificity of acyl-homoserine lactone signal production in bacterial quorum sensing. Mol Cell 9:685–694

    Article  CAS  PubMed  Google Scholar 

  • Yong YC, Zhong JJ (2010) N-acylated homoserine lactone production and involvement in the biodegradation of aromatics by an environmental isolate of Pseudomonas aeruginosa. Process Biochem 45:1944–1948

    Article  CAS  Google Scholar 

  • Yong YC, Zhong JJ (2013) Regulation of aromatics biodegradation by rhl quorum sensing system through induction of catechol meta-cleavage pathway. Bioresour Technol 136:761–765

    Article  CAS  PubMed  Google Scholar 

  • Zuloaga O, Prieto A, Usobiaga A, Sarkar S, Chatterjee M, Bhattacharya B, Bhattacharya A, Alam MA, Satpathy K (2009) Polycyclic aromatic hydrocarbons in intertidal marine bivalves of Sunderban mangrove wetland, India: an approach to bioindicator species. Water Air Soil Pollut 201:305–318

    Article  CAS  Google Scholar 

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Acknowledgments

The authors would like to thank the authorities of NIT, Rourkela for providing facilities. Financial support received from the Department of Biotechnology, Ministry of Science and Technology, Government of India on the research project on bioremediation by biofilm-forming marine bacteria is gratefully acknowledged. They are also thankful to Central Instrument Facility, Bose Institute, Kolkata for providing the GC-MS and HPLC facilities.

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The authors declare that they have no competing interests.

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  1. Laboratory of Environmental Microbiology and Ecology (LEnME), Department of Life Science, National Institute of Technology, Rourkela, Odisha, 769 008, India

    Neelam Mangwani, Supriya Kumari & Surajit Das

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  1. Neelam Mangwani
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Correspondence to Surajit Das.

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Mangwani, N., Kumari, S. & Das, S. Involvement of quorum sensing genes in biofilm development and degradation of polycyclic aromatic hydrocarbons by a marine bacterium Pseudomonas aeruginosa N6P6. Appl Microbiol Biotechnol 99, 10283–10297 (2015). https://doi.org/10.1007/s00253-015-6868-7

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