Advertisement

Folia Microbiologica

, Volume 57, Issue 6, pp 501–508 | Cite as

Structural characterization and surface activities of biogenic rhamnolipid surfactants from Pseudomonas aeruginosa isolate MN1 and synergistic effects against methicillin-resistant Staphylococcus aureus

  • Nasrin SamadiEmail author
  • Neda Abadian
  • Reza Ahmadkhaniha
  • Farzaneh Amini
  • Dina Dalili
  • Noushin Rastkari
  • Eliyeh Safaripour
  • Farzaneh Aziz Mohseni
Article

Abstract

The aim of present work was to study chemical structures and biological activities of rhamnolipid biosurfactants produced by Pseudomonas aeruginosa MN1 isolated from oil-contaminated soil. The results of liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis revealed that total rhamnolipids (RLs) contained 16 rhamnolipid homologues. Di-lipid RLs containing C10-C10 moieties were by far the most predominant congeners among mono-rhamnose (53.29 %) and di-rhamnose (23.52 %) homologues. Mono-rhamnolipids form 68.35 % of the total congeners in the RLs. Two major fractions were revealed in the thin layer chromatogram of produced RLs which were then purified by column chromatography. The retardation factors (R f) of the two rhamnolipid purple spots were 0.71 for RL1 and 0.46 for RL2. LC-MS/MS analysis proved that RL1 was composed of mono-RLs and RL2 consisted of di-RLs. RL1 was more surface-active with the critical micelle concentration (CMC) value of 15 mg/L and the surface tension of 25 mN/m at CMC. The results of biological assay showed that RL1 is a more potent antibacterial agent than RL2. All methicillin-resistant Staphylococcus aureus (MRSA) strains were inhibited by RLs that were independent of their antibiotic susceptibility patterns. RLs remarkably enhanced the activity of oxacillin against MRSA strains and lowered the minimum inhibitory concentrations of oxacillin to the range of 3.12–6.25 μg/mL.

Keywords

Critical Micelle Concentration Mineral Salt Medium Oxacillin Surface Tension Measurement Biosurfactant Production 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Abbreviations

CFU

Colony-forming unit

CMC

Critical micelle concentration

FIC

Fractional inhibitory concentration

LC-MS

Liquid chromatography-mass spectrometry

MHB

Mueller–Hinton broth

MIC

Minimum inhibitory concentration

MRSA

Methicillin-resistant Staphylococcus aureus

MSM

Mineral salts medium

PTCC

Persian type culture collection

Rf

Retardation factor

RL

Rhamnolipid

TIC

Total ion chromatogram

TLC

Thin layer chromatography

Notes

Acknowledgements

This study was supported by a grant from Tehran University of Medical Sciences (no. 5664).

References

  1. Abalos A, Pinazo A, Infante MR, Casals M, Garcia F, Manresa A (2001) Physiochemical and antimicrobial properties of new rhamnolipids produced by Pseudomonas aeruginosa AT10 from soybean oil refinery wastes. Langmuir 17:1367–1371CrossRefGoogle Scholar
  2. Abdel-Mawgoud AM, Lépine F, Déziel E (2010) Rhamnolipids: diversity of structures, microbial origins and roles. Appl Microbiol Biotechnol 86:1323–1336PubMedCrossRefGoogle Scholar
  3. Akbarzadeh T, Fallah Tafti A, Samadi N, Foroumadi A, Amanlou M, Faramarzi MA, Shafiee A (2010) Synthesis and cloxacillin antimicrobial enhancement of 2-methylsulfonylimidazolyl-1,4-dihydropyridine derivatives. Daru 18:118–123PubMedGoogle Scholar
  4. Chen SY, Wei YH, Chang JS (2007) Repeated pH-stat fed-batch fermentation for rhamnolipid production with indigenous Pseudomonas aeruginosa S2. Appl Microbiol Biotechnol 76:67–74PubMedCrossRefGoogle Scholar
  5. Cohen R, Exerowa D (2007) Surface forces and properties of foam films from rhamnolipid biosurfactants. Adv Colloid Interface 135:24–34CrossRefGoogle Scholar
  6. Desai JD, Banat IM (1997) Microbial production of surfactant and their commercial potential. Microbiol Mol Biol Rev 61:47–64PubMedGoogle Scholar
  7. Déziel E, Lépine F, Dennie D, Biosmenu D, Mamer OA, Villemur R (1999) Liquid chromatography/mass spectrometry analysis of mixtures of rhamnolipids produced by Pseudomonas aeruginosa strain 57RP grown on mannitol or naphtalene. Biochim Biophys Acta 1440:244–252PubMedCrossRefGoogle Scholar
  8. Déziel E, Lépine F, Milot S, Villemur R (2000) Mass spectrometry monitoring of rhamnolipids from growing culture of Pseudomonas aeruginosa strain from 57RP. Biochim Biophys Acta 1485:145–152PubMedCrossRefGoogle Scholar
  9. Emami S, Foroumadi A, Faramarzi MA, Samadi N (2008) Synthesis and Antibacterial activity of quinolone-based compounds containing a coumarin moiety. Arch Pharm Chem Life Sci 341:42–48CrossRefGoogle Scholar
  10. Emami S, Foroumadi A, Samadi N, Faramarzi MA, Rajabalian S (2009) Conformationally constrained analogs of n-substituted piperazinylquinolones: synthesis and antibacterial activity of N-(2,3-Dihydro-4-hydroxyimino-4 H-1-benzopyran-3-yl)-piperazinylquinolones. Arch Pharm Chem Life Sci 342:405–411CrossRefGoogle Scholar
  11. Glover RE, Smith RR, Jones MV, Jackson SK, Rowlands CC (1999) An EPR investigation of surfactant action on bacterial membranes. FEMS Microbiol Lett 177:57–62PubMedCrossRefGoogle Scholar
  12. Haba E, Pinazo A, Jauregui O, Espuny MJ, Infante MR, Manresa A (2003) Physicochemical characterization and antimicrobial properties of rhamnolipids produced by Pseudomonas aeruginosa 47 T2 NCBIM 40044. Biotechnol Bioeng 81:316–322PubMedCrossRefGoogle Scholar
  13. Jarvis FG, Johnson MJ (1949) A glyco-lipid produced by Pseudomonas aeruginosa. J Am Chem Soc 71:4124–4126CrossRefGoogle Scholar
  14. Khalaj A, Nakhjiri M, Negahbani AS, Samadizadeh M, Firoozpour L, Rajabalian S, Samadi N, Faramarzi MA, Adibpour N, Shafiee A, Foroumadi AR (2011) Discovery of a novel nitroimidazolyleoxazolidinone hybrid with potent anti gram-positive activity: synthesis and antibacterial evaluation. Eur J Med Chem 46:65–70PubMedCrossRefGoogle Scholar
  15. Krieg NR, Holt JG (2001) Bergey's manual of systematic bacteriology. Williams and Wilkins, BaltimoreGoogle Scholar
  16. Kumar KA, Mazumdar K, Dutta NK, Karak P, Dastidar SG, Ray R (2004) Evaluation of synergism between the aminoglycoside antibiotic streptomycin and the cardiovascular agent amlodipine. Biol Pharm Bull 27:1116–1120CrossRefGoogle Scholar
  17. Kurup TR, Wan LS, Chan LW (1991) Effect of surfactants on the antibacterial activity of preservatives. Pharm Acta Helv 66:274–280PubMedGoogle Scholar
  18. Lang S, Wullbrandt D (1999) Rhamnose lipids-biosynthesis, microbial production and application potential. Appl Microbiol Biotechnol 51:22–23PubMedCrossRefGoogle Scholar
  19. Lee J, Choi Y, Woo ER, Lee DG (2009) Antibacterial and synergistic activity of isocryptomerin isolated from Selaginella tamariscina. J Microbiol Biotechnol 19:204–207PubMedCrossRefGoogle Scholar
  20. Lindhardt TJ, Bakhit R, Danies L, Mayerl R, Pickenhagen W (1989) Microbially produced rhamnolipid as a source of rhamnose. Biotechnol Bioeng 33:365–368CrossRefGoogle Scholar
  21. Lotfabad TB, Abassi H, Ahmadkhaniha R, Roostaazad R, Masoomi F, Zahiri HS, Ahmadian G, Vali H, Noghabi KA (2010) Structural characterization of a rhamnolipid-type biosurfactant produced by Pseudomonas aeruginosa MR01: enhancement of di-rhamnolipid proportion using gamma irradiation. Colloids Surf B 81:397–405CrossRefGoogle Scholar
  22. Maier RM, Soberon-Chavez G (2000) Pseudomonas aeruginosa rhamnolipids: biosynthesis and potential applications. Appl Microbiol Biotechnol 54:625–633PubMedCrossRefGoogle Scholar
  23. Mata-Sandoval JC, Karns J, Torrents A (1999) High-performance liquid chromatography method for the characterization of rhamnolipid mixtures produce by Pseudomonas aeruginosa UG2 on corn oil. J Chromatogr A 864:211–220PubMedCrossRefGoogle Scholar
  24. Mata-Sandoval JC, Karns J, Torrents A (2000) Effect of nutritional and environmental conditions on the production and composition of rhamnolipids by Pseudomonas aeruginosa UG2. Microbiol Res 155:1–8CrossRefGoogle Scholar
  25. Menichetti F (2005) Current and emerging serious gram-positive infections. Clin Microbiol Infec 11:22–28CrossRefGoogle Scholar
  26. Mulligan CN (2005) Environmental applications for biosurfactants. Environ Pollut 133:183–198PubMedCrossRefGoogle Scholar
  27. Muroi H, Nihei KI, Tsujimoto K, Kubo I (2004) Synergistic effects of anacardic acids and methicillin against methicillin resistant Staphylococcus aureus. Bioorg Med Chem 12:583–587PubMedCrossRefGoogle Scholar
  28. NCCLS (2006) Methods for Dilution antimicrobial susceptibility tests for bacteria that grow aerobically. approved standard M7-A7, 7th edition, Wayne, Pennsylvania.Google Scholar
  29. Nielsen L, Kadavy D, Rajagopal S, Drijber R, Kenneth W (2005) Survey of extreme solvent tolerance in gram-positive cocci: membrane fatty acid changes in Staphylococcus haemolyticus grown in toluene. Appl Environ Microbiol 71:5171–5176PubMedCrossRefGoogle Scholar
  30. Nitschke M, Costa SGVAO, Contiero J (2005) Rhamnolipid surfactants: an update on the general aspects of these remarkable biomolecules. Biotechnol Progr 21:1593–1600CrossRefGoogle Scholar
  31. Özdemir G, Peker S, Helvaci ŞŞ (2004) Effect of pH on the surface and interfacial behavior of rhamnolipids R1 and R2. Colloid Surface Physicochem Eng Aspect 234:135–143CrossRefGoogle Scholar
  32. Pashynska VA (2009) Mass spectrometric study of rhamnolipid biosurfactants and their interactions with cell membrane phospholipids. Biopolym Cell 25:504–508Google Scholar
  33. Patrone V, Campana R, Vittoria E, Baffone W (2010) In vitro synergistic activities of essential oils and surfactants in combination with cosmetic preservatives against Pseudomonas aeruginosa and Staphylococcus aureus. Curr Microbiol 60:237–241PubMedCrossRefGoogle Scholar
  34. Rendell NB, Taylor GW, Somerville M, Todd H, Wilson R, Cole P (1990) Characterization of Pseudomonas rhamnolipids. Biochim Biophys Acta 1045:189–193PubMedCrossRefGoogle Scholar
  35. Rosenberg E, Ron EZ (1999) High and low molecular-mass microbial surfactants. Appl Microbiol Biotechnol 52:154–162PubMedCrossRefGoogle Scholar
  36. Sadowska B, Walencka E, Wieckowska-Szakiel M, Różalska B (2010) Bacteria competing with the adhesion and biofilm formation by Staphylococcus aureus. Folia Microbiol 55:497–501CrossRefGoogle Scholar
  37. Salimnia H, Brown WJ (2005) Detection of oxacillin resistance in Staphylococcus aureus: comparison of phoenix oxacillin and cefoxitin MICs, microscan oxacillin MIC, oxacillin and cefoxitin disk diffusion, and mecA gene detection. ICAAC 24:12–20Google Scholar
  38. Sook KE, Jeong SI, Kim JH, Park C, Kim SM, Kim JK, Lee KM, Lee SH, So H, Park R (2009) Synergistic effects of the combination of 20-hydroxyecdysone with ampicillin and gentamicin against methicillin-resistant Staphylococcus aureus. J Microbiol Biotechnol 19:1576–1581Google Scholar
  39. Sotirova AV, Spasova DI, Galabova DN, Karpenko E, Shulga A (2008) Rhamnolipid biosurfactant permeabilizing effects on gram-positive and gram-negative bacterial strains. Curr Microbiol 56:639–644PubMedCrossRefGoogle Scholar
  40. Velho RV, Medina LFC, Segalin J, Brandelli A (2011) Production of lipopeptides among Bacillus strains showing growth inhibition of phytopathogenic fungi. Folia Microbiol 56:297–303CrossRefGoogle Scholar
  41. Youssef NH, Dunacn KE, Nagle DP, Savage KN, Knapp RM, McInerney MJ (2004) Comparison of methods to detect biosurfactant production by diverse microorganism. J Microbiol Meth 56:339–47CrossRefGoogle Scholar

Copyright information

© Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i. 2012

Authors and Affiliations

  • Nasrin Samadi
    • 1
    Email author
  • Neda Abadian
    • 1
  • Reza Ahmadkhaniha
    • 2
  • Farzaneh Amini
    • 1
  • Dina Dalili
    • 1
  • Noushin Rastkari
    • 3
  • Eliyeh Safaripour
    • 4
  • Farzaneh Aziz Mohseni
    • 5
  1. 1.Department of Drug and Food Control and Biotechnology Research Center, Faculty of PharmacyTehran University of Medical SciencesTehranIran
  2. 2.Pharmaceutical Sciences Research CenterTehran University of Medical SciencesTehranIran
  3. 3.Institute for Environmental ResearchTehran University of Medical SciencesTehranIran
  4. 4.Pharmaceutical Quality Assurance Research Center, Faculty of PharmacyTehran University of Medical SciencesTehranIran
  5. 5.Biotechnology DepartmentIranian Research Organization for Science and Technology (IROST)TehranIran

Personalised recommendations