Abstract
Biofilm associated infections are the major contributor of mortality, morbidity and financial burden in patients with a bacterial infection. About 65% of all bacterial infections are associated with the information of bacterial biofilms. Bacterial biofilms not only reduce the efficacy of antibacterial treatment but also increases the threat of developing antibacterial resistance. Recently, our group has discovered the antibacterial activity of Fmoc-phenylalanine (Fmoc-F) and other Fmoc-amino acids (Fmoc-AA). Fmoc-F and other Fmoc-AA showed antibacterial activity due to their surfactant properties. Surfactants are known to eradicate biofilm and enhance antimicrobial activity in biofilm. Thus, in the present study, we evaluated the anti-biofilm activity of Fmoc-F against clinically relevant bacteria. We found that Fmoc-F not only inhibits the biofilm formation in Staphylococcus aureus and Pseudomonas aeruginosa, but also eradicates the already formed biofilms over the surface. Further, Fmoc-F coated glass surface resists S. aureus and P. aeruginosa biofilm formation and attachment, when biofilm is grown over the surface. The mechanistic investigation suggests that Fmoc-F reduces the extracellular matrix (ECM) components such as proteins, carbohydrates and eDNA in the biofilm and affect its stability via direct interactions with ECM components and/ or indirectly through reducing bacterial cell population. Finally, we showed that Fmoc-F treatment in combination with vancomycin and ampicillin synergistically inhibit biofilm formation. Overall, the study demonstrates the potential application of Fmoc-F and other Fmoc-AA molecules individually as well as in combination as anti-biofilm coating material for treating biofilm associated infections.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Zhang W, et al. Extracellular matrix-associated proteins form an integral and dynamic system during Pseudomonas aeruginosa biofilm development. Front Cell Infect Microbiol. 2015;5:40.
Berlanga M, Guerrero R. Living together in biofilms: the microbial cell factory and its biotechnological implications. Micro Cell Fact. 2016;15:165.
Jamal M, et al. Bacterial biofilm and associated infections. J Chin Med Assoc. 2018;81:7–11.
Aparna MS, Yadav S. Biofilms: microbes and disease. Braz J Infect Dis. 2008;12:526–30.
Khatoon Z, McTiernan CD, Suuronen EJ, Mah T-F, Alarcon EI. Bacterial biofilm formation on implantable devices and approaches to its treatment and prevention. Heliyon 2018;4:e01067.
Algburi A, Comito N, Kashtanov D, Dicks LMT, Chikindas ML. Control of Biofilm Formation: Antibiotics and Beyond. Appl Environ Micro. 2017;83:e02508–16.
Kvist M, Hancock V, Klemm P. Inactivation of efflux pumps abolishes bacterial biofilm formation. Appl Environ Micro. 2008;74:7376–82.
Zhang L, Mah T-F. Involvement of a novel efflux system in biofilm-specific resistance to antibiotics. J Bacteriol. 2008;190:4447–52.
Gebreyohannes G, Nyerere A, Bii C, Sbhatu DB. Challenges of intervention, treatment, and antibiotic resistance of biofilm-forming microorganisms. Heliyon 2019;5:e02192.
Kalpana BJ, Aarthy S, Pandian SK. Antibiofilm activity of α-amylase from Bacillus subtilis S8-18 against biofilm forming human bacterial pathogens. Appl Biochem Biotechnol. 2012;167:1778–94.
Kalishwaralal K, BarathManiKanth S, Pandian SRK, Deepak V, Gurunathan S. Silver nanoparticles impede the biofilm formation by Pseudomonas aeruginosa and Staphylococcus epidermidis. Colloids Surf B Biointerfaces. 2010;79:340–4.
Kolodkin-Gal I, et al. D-amino acids trigger biofilm disassembly. Science. 2010;328:627–9.
Hochbaum AI, et al. Inhibitory effects of D-amino acids on Staphylococcus aureus biofilm development. J Bacteriol. 2011;193:5616–22.
McCloskey AP, Draper ER, Gilmore BF, Laverty G. Ultrashort self-assembling Fmoc-peptide gelators for anti-infective biomaterial applications. J Pept Sci. 2017;23:131–40.
Gahane AY, et al. Fmoc-phenylalanine displays antibacterial activity against Gram-positive bacteria in gel and solution phases. Soft Matter. 2018;14:2234–44.
Tyldesley HC, Salisbury A, Chen R, Mullin M, Percival SL. Surfactants and their role in biofilm management in chronic wounds. Wounds Int. 2019;10:20–4.
Percival SL, et al. Surfactants: role in biofilm management and cellular behaviour. Int Wound J 2019;16:753–60.
Das Ghatak P, Mathew-Steiner SS, Pandey P, Roy S, Sen CK. A surfactant polymer dressing potentiates antimicrobial efficacy in biofilm disruption. Sci Rep. 2018;8:873.
O’Toole GA. Microtiter dish biofilm formation assay. J Vis Exp. 2011;47:2437.
Chiba A, Sugimoto S, Sato F, Hori S, Mizunoe Y. Extraction of ECM from bacterial biofilms. Micro Biotechnol. 2015;8:392–403.
Mak YM, Ho KK. An improved method for the isolation of chromosomal DNA from various bacteria and cyanobacteria. Nucleic Acids Res. 1992;20:4101–2.
Alhede M, Jensen PØ, Givskov M, Thomas B Biofilm of medical importance, vol. XII. Eolss Publishers, 2009.
Percival SL, Mayer D, Salisbury A-M. Efficacy of a surfactant-based wound dressing on biofilm control. Wound Repair Regen. 2017;25:767–73.
Simões M, Simões LC, Pereira MO, Vieira MJ. Sodium dodecyl sulfate allows the persistence and recovery of biofilms of Pseudomonas fluorescens formed under different hydrodynamic conditions. Biofouling 2008;24:35–44.
Chandra N, Tyagi VK. Synthesis, properties, and applications of amino acids based surfactants: a review. J Dispers Sci Technol. 2013;34:800–8.
Singh V, Snigdha K, Singh C, Sinha N, Thakur AK. Understanding the self-assembly of Fmoc–phenylalanine to hydrogel formation. Soft Matter. 2015;11:5353–64.
Yuan Y, Hays MP, Hardwidge PR, Kim J. Surface characteristics influencing bacterial adhesion to polymeric substrates. RSC Adv. 2017;7:14254–61.
Koo H, Yamada KM. Dynamic cell-matrix interactions modulate microbial biofilm and tissue 3D microenvironments. Curr Opin Cell Biol. 2016;42:102–12.
Steinberg N, Kolodkin-Gal I. The matrix reloaded: how sensing the extracellular matrix synchronizes bacterial communities. J Bacteriol. 2015;197:2092.
Karadenizli A, Kolayli F, Ergen K. A novel application of Fourier-transformed infrared spectroscopy: classification of slime from staphylococci. Biofouling 2007;23:63–71.
Flemming H-C, Wingender J. The biofilm matrix. Nat Rev Microbiol. 2010;8:623–33.
Dueholm MS, et al. Expression of Fap amyloids in Pseudomonas aeruginosa, P. fluorescens, and P. putida results in aggregation and increased biofilm formation. Microbiologyopen 2013;2:365–82.
Zeng G, et al. Functional bacterial amyloid increases Pseudomonas biofilm hydrophobicity and stiffness. Front Microbiol. 2015;6:1099.
Schwartz K, Syed AK, Stephenson RE, Rickard AH, Boles BR. Functional amyloids composed of phenol soluble modulins stabilize Staphylococcus aureus biofilms. PLoS Pathog. 2012;8:e1002744.
Van Gerven N, Van der Verren SE, Reiter DM, Remaut H. The role of functional amyloids in bacterial virulence. J Mol Biol. 2018;430:3657–84.
Yakupova EI, Bobyleva LG, Vikhlyantsev IM, Bobylev AG. Congo red and amyloids: history and relationship. Biosci Rep. 2019;39:BSR20181415.
Xue C, Lin TY, Chang D, Guo Z. Thioflavin T as an amyloid dye: fibril quantification, optimal concentration and effect on aggregation. R Soc Open Sci. 2017;4:160696.
Biancalana M, Koide S. Molecular mechanism of Thioflavin-T binding to amyloid fibrils. Biochim Biophys Acta. 2010;1804:1405–12.
Xu X, et al. Synergistic combination of two antimicrobial agents closing each other’s mutant selection windows to prevent antimicrobial resistance. Sci Rep. 2018;8:7237.
Marquès C, et al. Effects of antibiotics on biofilm and unattached cells of a clinical Staphylococcus aureus isolate from bone and joint infection. J Med Microbiol. 2015;64:1021–6.
Tamma PD, Cosgrove SE, Maragakis LL. Combination therapy for treatment of infections with gram-negative bacteria. Clin Microbiol Rev. 2012;25:450–70.
Acknowledgements
We authors thank IITK, CSIR and MHRD, Government of India for funding the fellowships. This work was supported financially by the Indian Institute of Technology Kanpur, MHRD India (Project No. IITK/BSBE/20100293).
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary information
Rights and permissions
About this article
Cite this article
Singh, H., Gahane, A., Singh, V. et al. Antibiofilm activity of Fmoc-phenylalanine against Gram-positive and Gram-negative bacterial biofilms. J Antibiot 74, 407–416 (2021). https://doi.org/10.1038/s41429-021-00409-2
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/s41429-021-00409-2