Startseite Enhanced biofilm disruption in ESKAPE pathogens through synergistic activity of EPS degrading enzymes
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Enhanced biofilm disruption in ESKAPE pathogens through synergistic activity of EPS degrading enzymes

  • Pooja Rao , Jamuna B. Aswathanarayan ORCID logo EMAIL logo , SubbaRao V. Madhunapantula , Ravishankar V. Rai , Ponnadurai Ramasami und Sowmya G. Shivappa
Veröffentlicht/Copyright: 8. April 2025
Pure and Applied Chemistry
Aus der Zeitschrift Pure and Applied Chemistry Band 97 Heft 5

Abstract

Biofilm formation by Klebsiella pneumoniae, Acinetobacter baumannii, and methicillin-resistant Staphylococcus aureus (MRSA) significantly contributes to antimicrobial resistance (AMR), complicating infections associated with medical devices. This study investigates the potential of α-Amylase, DNase I, and Proteinase K in disrupting extracellular polymeric substances (EPS) within biofilms to enhance biofilm inhibition. Molecular docking studies revealed a strong interaction between α-Amylase and cellulose (−3.58 kcal/mol), suggesting effective targeting of biofilm polysaccharides. Biofilm inhibition was quantified using the crystal violet microtiter plate assay, and structural changes were visualized through confocal laser scanning microscopy (CLSM). The checkerboard synergy assay showed that enzyme combinations achieved up to 90 % biofilm inhibition, significantly outperforming individual enzymes (50–70 %). Notably, α-Amylase + DNase I and α-Amylase + Proteinase K exhibited synergy (FICI ≤ 0.5) in MRSA and A. baumannii, while DNase I + Proteinase K showed limited activity in A. baumannii (FICI = 2.0), suggesting biofilm composition differences influence enzymatic activity. Confocal microscopy analysis revealed that biofilm thickness was reduced by 50 % in K. pneumoniae and by 60 % in A. baumannii, further supporting enzyme-mediated biofilm disruption. These findings highlight the potential clinical applications of enzymatic therapy, particularly in preventing ventilator-associated pneumonia (VAP) by inhibiting biofilm formation and disrupting preformed biofilms in endotracheal tubes and improving antimicrobial efficacy. This study establishes α-Amylase as a potent biofilm-disrupting agent, with synergistic enzyme therapy offering a viable strategy to combat biofilm-associated infections. Currently, we are focusing on optimizing enzyme formulations for clinical application and evaluating their combination with antibiotics to enhance therapeutic outcomes.

Keywords: α-amylase; biofilm inhibition; ESKAPE pathogens; extracellular polymeric substances (EPS); molecular docking; synergistic activity; VCCA-2024
Corresponding author: Jamuna B. Aswathanarayan, Department of Microbiology, JSS Academy of Higher Education and Research, Mysuru, India, e-mail:
Article note: A collection of invited papers based on presentations at the Virtual Conference on Chemistry and its Applications held on 12–16 August 2024.

Funding source: UGC-BSR Research Start-Up Grant

Award Identifier / Grant number: No.F.30-601/2021 (BSR) dated 15th March 2023

Acknowledgments

We extend our sincere thanks to the Center of Excellence in Molecular Biology and Regenerative Medicine (CEMR) laboratory for their assistance with the confocal microscopy studies (Leica Stellaris-5 Confocal Laser Scanning Microscope- Funded by DST PURSE project-SR/PURSE/2021/81-(C) Year: 2022). We are grateful to Special Interest group - Biofilms in Clinical Settings and their Control, JSS AHER and UGC-BSR Research Start-up Grant for funding this study

  1. Research ethics: Not applicable.

  2. Informed consent: Not applicable.

  3. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  4. Use of Large Language Models, AI and Machine Learning Tools: None declared.

  5. Conflict of interest: The authors state no conflict of interest.

  6. Research funding: “Special Interest group- Biofilms in Clinical Settings and their Control. No: REG/FLS/GEN/2020-21/155 dated 14/07/2020” and “UGC-BSR Research Start-up Grant (No.F.30-601/2021 (BSR) dated 15th March 2023)”.

  7. Data availability: Not applicable.

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Published Online: 2025-04-08
Published in Print: 2025-05-26

© 2025 IUPAC & De Gruyter

Artikel in diesem Heft

  1. Frontmatter
  2. In this issue
  3. Research Articles
  4. Efficient degradation of 1,2-dichlorobenzene using heterogeneous catalytic ozonation over metal loaded gamma alumina catalysts
  5. Effect of chemical modification using glyoxylic acid on the stability of α-amylase from Aspergillus fumigatus
  6. A new, innovative, simple method to determine the concentration of phosphate and sulphate ions in an aqueous extract of plants using conductometric titration
  7. Metalloporphyrin-mediated oxidative degradation of risperidone under mild conditions: an LC-MS/MS study
  8. Synthesis of novel ternary herbicide-layered double hydroxide hybrids via the ion exchange method
  9. Water Quality Index and the quality of freshwater resource uMhlathuze river, Kwazulu-Natal, South Africa: A Review
  10. Experimental ‘in-Vitro’ investigation on bio-chemical constituents, radical scavenging activity, and reducing power assay of cow urine
  11. Enhanced biofilm disruption in ESKAPE pathogens through synergistic activity of EPS degrading enzymes
  12. Green synthesis of silver nanoparticles using a bioflocculant produced by a kombucha tea yeast isolate for antimicrobial and biosafety testing
  13. Characterization of metabolite compounds from endophytic fungi associated with white turi plant (Sesbania grandiflora) and their antibacterial activity
  14. Enhanced photocatalytic degradation of methylene blue dye using TiO2 nanoparticles obtained via chemical and green synthesis: a comparative analysis
  15. Characterization of electrocatalysts for the oxygen evolution reaction OER: a bi-metal study IrM oxides (Ru, and Au)
  16. Development of a second harmonic generation microscope optimized for biomaterial studies
https://doi.org/10.1515/pac-2024-0399
Schlagwörter für diesen Artikel
α-amylase; biofilm inhibition; ESKAPE pathogens; extracellular polymeric substances (EPS); molecular docking; synergistic activity; VCCA-2024

Artikel in diesem Heft

  1. Frontmatter
  2. In this issue
  3. Research Articles
  4. Efficient degradation of 1,2-dichlorobenzene using heterogeneous catalytic ozonation over metal loaded gamma alumina catalysts
  5. Effect of chemical modification using glyoxylic acid on the stability of α-amylase from Aspergillus fumigatus
  6. A new, innovative, simple method to determine the concentration of phosphate and sulphate ions in an aqueous extract of plants using conductometric titration
  7. Metalloporphyrin-mediated oxidative degradation of risperidone under mild conditions: an LC-MS/MS study
  8. Synthesis of novel ternary herbicide-layered double hydroxide hybrids via the ion exchange method
  9. Water Quality Index and the quality of freshwater resource uMhlathuze river, Kwazulu-Natal, South Africa: A Review
  10. Experimental ‘in-Vitro’ investigation on bio-chemical constituents, radical scavenging activity, and reducing power assay of cow urine
  11. Enhanced biofilm disruption in ESKAPE pathogens through synergistic activity of EPS degrading enzymes
  12. Green synthesis of silver nanoparticles using a bioflocculant produced by a kombucha tea yeast isolate for antimicrobial and biosafety testing
  13. Characterization of metabolite compounds from endophytic fungi associated with white turi plant (Sesbania grandiflora) and their antibacterial activity
  14. Enhanced photocatalytic degradation of methylene blue dye using TiO2 nanoparticles obtained via chemical and green synthesis: a comparative analysis
  15. Characterization of electrocatalysts for the oxygen evolution reaction OER: a bi-metal study IrM oxides (Ru, and Au)
  16. Development of a second harmonic generation microscope optimized for biomaterial studies
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