Skip to main content
SpringerLink
Log in

Analysis of Pseudomonas aeruginosa PAO1 Biofilm Protein Profile After Exposure to n-Butanolic Cyclamen coum Extract Alone and in Combination with Ciprofloxacin

  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

Pseudomonas aeruginosa biofilm-related infections are the major cause of premature death in cystic fibrosis patients. Strategies to induce biofilm dispersal are of interest, because of their potential in preventing biofilm-related infections. Our previous work demonstrated that n-butanolic Cyclamen coum extract with ciprofloxacin could eliminate 1- and 3-day-old P. aeruginosa PAO1 biofilms. To gain new insights into the role of C. coum extract and its synergistic effect with ciprofloxacin in eliminating P. aeruginosa PAO1 biofilms, two-dimensional gel electrophoresis (2-DE) in combination with mass spectrometry-based protein identification were used. Changes in the bacterial protein expression were analyzed when 3-day-old biofilm cells were exposed to the C. coum extract alone and in combination with ciprofloxacin. Proteins involved in alginate biosynthesis, quorum sensing, adaptation/protection, carbohydrate and amino acid metabolism showed a weaker expression in the C. coum extract-ciprofloxacin-treated biofilm cells compared to those in the untreated cells. Interestingly, the proteome of C. coum extract-ciprofloxacin-treated biofilm revealed more resemblance to the planktonic phenotype than to the biofilm phenotype. It appears that saponin extract in combination with ciprofloxacin causes biofilm disruption due to several mechanisms such as motility induction, cell envelope integrity perturbation, stress protein expression reduction, and more importantly, signal transduction perturbation. In conclusion, exposure to a combination of biofilm dispersal such as saponin extract and antimicrobial agents may offer a novel strategy to control preestablished, persistent P. aeruginosa biofilms and biofilm-related infections.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

Abbreviations

2-DE:

Two-dimensional gel electrophoresis

TSA:

Tryptic soy agar

TSB:

Trypticase soy broth

IPG:

Immobilized pH gradient

PMSF:

Ammonium persulfate and phenylmethylsulfonylfluoride

THU:

Tehran Herbarium University

DMSO:

Dimethyl-sulphoxide

FBECs:

Fractional biofilm eliminating concentration

MBEC:

Minimum biofilm eliminating concentration

BSA:

Bovine serum albumin

ACN:

Acetonitrile

CAM:

Carbamidomethyl

Me:

Methionine

FA:

Formic acid

SD:

Standard deviations

Fgt:

Flagellar glycosyl transferase

Ndk:

Nucleoside diphosphate kinase

AHL:

Acyl-homoserine lactone

References

  1. Høiby, N., Ciofu, O., & Bjarnsholt, T. (2010). Pseudomonas aeruginosa biofilms in cystic fibrosis. Future Microbiology, 5, 1663–1674.

    Article  Google Scholar 

  2. Davies, D. (2003). Understanding biofilm resistance to antibacterial agents. Nature Reviews of Drug Discovery, 2, 114–122.

    Article  CAS  Google Scholar 

  3. Kaplan, J. B. (2010). Biofilm dispersal: mechanisms, clinical implications, and potential therapeutic uses. Journal of Dental Research, 89, 205–218.

    Article  CAS  Google Scholar 

  4. Nyakudya, E., Jeong, J. H., Lee, N. K., & Jeong, Y. S. (2014). Platycosides from the roots of Platycodon grandiflorum and their health benefits. Preventive Nutrition and Food Science, 19, 59–68.

    Article  CAS  Google Scholar 

  5. Shafiei, M., Abdi-Ali, A., Shahcheraghi, F., Saboora, A., & Akbari-Noghabi, K. (2014). Eradication of Pseudomonas aeruginosa biofilms using the combination of n-butanolic Cyclamen coum extract and ciprofloxacin. Jundishapur Journal of Microbiology, 7, e17430.

    Article  Google Scholar 

  6. Ahmadbeigi, Z., & Saboora, A. (2009). Comparison of three methods for saponin extraction from tuber of Cyclamen coum Miller. Journal of Science, 21, 27–38.

    Google Scholar 

  7. Hassan, S. M., Al- Aqil, A. A., & Attimarad, M. (2013). Determination of crude saponin and total flavonoids content in guar meal. Advancement in Medicinal Plant Research, 1, 24–28.

  8. Pennsylvania, W. (2007). Clinical and Laboratory Standards Institute, CLSI (Clinical and Laboratory Standards Institute). Performance standards for antimicrobial susceptibility testing, approved standard M100-S17. 17th Informational Supplement.

  9. Pitts, B., Hamilton, M. A., Zelver, N., & Stewart, P. S. (2003). A microtiter plate screening method for biofilm disinfection and removal. Journal of Microbiological Methods, 54, 269–276.

    Article  CAS  Google Scholar 

  10. Oosthuizen, M. C., Steyn, B., Theron, J., Cosette, P., Lindsay, D., Holy, A. V., & Brozel, V. S. (2002). Proteomic analysis reveals differential protein expression by Bacillus cereus during biofilm formation. Applied and Environmental Microbiology, 68, 2770–2780.

    Article  CAS  Google Scholar 

  11. Molloy, M.P., Herbert, B.R., Walsh, B.J., Tyler, M.I., Traini, M., Sanchez, J.C., Hochstrasser, D.F., Williams, K.L., Gooley., A.A. (1998). Extraction of membrane proteins by differential solubilization for separation using two- dimensional gel electrophoresis. Electrophoresis, 19(5), 837–44.

  12. Blum, H., Beier, H., & Gross, H. J. (1987). Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis, 8, 93–99.

    Article  CAS  Google Scholar 

  13. Masoudzadeh, N., Alidoust, L., Samie, N., Hajfarajollaha, H., Sharafi, H., Modiria, S., Zahiri, S. H., Vali, H., & Noghabi, K. A. (2014). Distinctive protein expression patterns of the strain Brevundimonas sp. ZF12 isolated from the aqueous zone containing high levels of radiation to cadmium-induced stress. Journal of Biotechnology, 186, 49–57.

    Article  CAS  Google Scholar 

  14. Vyshnava, S. S., Kanderi, D. P., Panjala, S. P., Pandian, P., Bontha, R. R., Goukanapalle, P. K. R., & Banaganapalli, B. (2016). Effect of silver nanoparticles against the formation of biofilm by Pseudomonas aeruginosa an in silico approach. Applied Biochemistry and Biotechnology. doi:10.1007/s12010-016-2107-7.

    Google Scholar 

  15. Izano, E., Amarante, M., Kher, W., & Kaplan, J. (2008). Differential roles of poly-N-acetylglucosamine surface polysaccharide and extracellular DNA in Staphylococcus aureus and Staphylococcus epidermidis biofilms. Applied and Environmental Microbiology, 74, 470–476.

    Article  CAS  Google Scholar 

  16. Goldberg, J., Hatano, K., & Pier, G. B. (1993). Synthesis of lipopolysaccharide-O-side-chains by Pseudomonas aeruginosa PAO1 requires the enzyme phosphomannomutase. Journal of Bacteriology, 175, 1605–1611.

    Article  CAS  Google Scholar 

  17. De Vos, D., Jr, A. L., Pirnay, J. P., Struelens, M., Vandenvelde, C., Duinslaeger, L., Vanderkelen, A., & Cornelis, P. (1997). Direct detection and identification of Pseudomonas aeruginosa in clinical samples such as skin biopsy specimens and expectorations by multiplex PCR based on two outer membrane lipoprotein genes, oprI and oprL. Journal of Clinical Microbiology, 35, 1295–1299.

    CAS  Google Scholar 

  18. Sauer, K., Camper, A., Ehrlich, G., & Costerton, J. D. (2002). Pseudomonas aeruginosa displays multiple phenotypes during development as a biofilm. Journal of Bacteriology, 184, 1140–1154.

    Article  CAS  Google Scholar 

  19. Stoodley, P., Hall-Stoodley, L., Boyle, J. D., Lappin-Scott, H. M., & Costerton, J. W. (2001). Growth and detachment of cell clusters from mature mixed-species biofilms. Applied and Environmental Microbiology, 67, 5608–5613.

    Article  CAS  Google Scholar 

  20. Clarke, M., Hughes, D., Zhu, C., Boedeker, E., & Sperandio, V. (2006). The QseC sensor kinase: a bacterial adrenergic receptor. Proceedings of the National Academy of Sciences, 103, 10420–10425.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The corresponding author would like to thank the National Institute of Genetic Engineering and Biotechnology (NIGEB) and the Vice Chancellor of the Alzahra University for financial support.

Author information

Authors and Affiliations

  1. Department of Microbiology, Faculty of Biological Sciences, Alzahra University, Tehran, Iran

    Morvarid Shafiei & Ahya Abdi-Ali

  2. Department of Bacteriology, Pasteur Institute of Iran, Tehran, Iran

    Morvarid Shafiei & Fereshteh Shahcheraghi

  3. Facility for Electron Microscopy Research, McGill University, 3640 Street, Montreal, Canada

    Hojatollah Vali

  4. Division of Industrial & Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14155-6343, Tehran, Iran

    Hossein Shahbani Zahiri & Kambiz Akbari Noghabi

Authors
  1. Morvarid Shafiei
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ahya Abdi-Ali
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Fereshteh Shahcheraghi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hojatollah Vali
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hossein Shahbani Zahiri
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Kambiz Akbari Noghabi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Kambiz Akbari Noghabi.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Shafiei, M., Abdi-Ali, A., Shahcheraghi, F. et al. Analysis of Pseudomonas aeruginosa PAO1 Biofilm Protein Profile After Exposure to n-Butanolic Cyclamen coum Extract Alone and in Combination with Ciprofloxacin. Appl Biochem Biotechnol 182, 1444–1457 (2017). https://doi.org/10.1007/s12010-017-2409-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-017-2409-4

Keywords

Search

Navigation