Abstract
Plant extracts have been used to treat microbiological diseases for centuries. This study examined plant triterpenoids tormentic acid (TA) and 23-hydroxycorosolic acid (HCA) for their antibiofilm effects on Staphylococcus aureus strains (MTCC-96 and MTCC-7405). Biofilms are bacterial colonies bound by a matrix of polysaccharides, proteins, and DNA, primarily impacting healthcare. As a result, ongoing research is being conducted worldwide to control and prevent biofilm formation. Our research showed that TA and HCA inhibit S. aureus planktonic growth by depolarizing the bacterial membrane. In addition, zone of inhibition studies confirmed their effectiveness, and crystal violet staining and biofilm protein quantification confirmed their ability to prevent biofilm formation. TA and HCA exhibited substantial reductions in biofilm formation for S. aureus (MTCC-96) by 54.85% and 48.6% and for S. aureus (MTCC-7405) by 47.07% and 56.01%, respectively. Exopolysaccharide levels in S. aureus biofilm reduced significantly by TA (25 μg/mL) and HCA (20 μg/mL). Microscopy, bacterial motility, and protease quantification studies revealed their ability to reduce motility and pathogenicity. Furthermore, TA and HCA treatment reduced the mRNA expression of S. aureus virulence genes. In silico analysis depicted a high binding affinity of triterpenoids for biofilm and quorum-sensing associated proteins in S. aureus, with TA having the strongest affinity for TarO (– 7.8 kcal/mol) and HCA for AgrA (– 7.6 kcal/mol). TA and HCA treatment reduced bacterial load in S. aureus-infected peritoneal macrophages and RAW264.7 cells. Our research indicates that TA and HCA can effectively combat S. aureus by inhibiting its growth and suppressing biofilm formation.
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Data availability
The authors have stated that the datasets produced and analyzed in the course of the present study are accessible upon reasonable request.
Abbreviations
- TA:
-
Tormentic acid
- HCA:
-
23-Hydroxycorosolic acid
- S. aureus :
-
Staphylococcus aureus
- LB:
-
Luria broth
- MIC:
-
Minimum inhibitory concentration
- CV:
-
Crystal violet
- Str:
-
Streptomycin
- AO:
-
Acridine orange
- DiSC3(5):
-
3,3′-Dipropylthiadicarbocyanine iodide
- DMEM:
-
Dulbecco’s Modified Eagle Medium
- FBS:
-
Fetal bovine serum
- PCR:
-
Polymerase chain reaction
- RNA:
-
Ribonucleic acid
- DNA:
-
Deoxyribonucleic acid
- cDNA:
-
Complementary DNA
- mRNA:
-
Messenger RNA
- QS:
-
Quorum sensing
- CFU:
-
Colony-forming unit
- EPS:
-
Extra polymeric substances
- ADT:
-
AutoDock Tools
- MOI:
-
Multiplicity of infection
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Acknowledgements
The authors express their sincere gratitude to the State Biotech Hub at Tripura University and Dr. Syed Arshad Hussain from the Department of Physics for generously providing access to the instrumental facility. The research conducted in Dr. Bhattacharjee's laboratory was supported by grants received from the Science and Engineering Research Board (DST-SERB) India, the Department of Biotechnology (DBT) India, and the Indian Council of Medical Research (ICMR) India.
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CG—Designed and performed experiments analyzed data and co- wrote the paper; SB, MCD, SA, JB, and RG—Performed experiments; PS, MK and YA—Performed bioinformatics analysis; BB—performed microscopy analysis; UCD—Designed experiments; and SB—Designed experiments, co-ordinate and supervised overall research and co-wrote the paper. All authors reviewed the manuscript.
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Ghosh, C., Das, M.C., Acharjee, S. et al. Combating Staphylococcus aureus biofilm formation: the inhibitory potential of tormentic acid and 23-hydroxycorosolic acid. Arch Microbiol 206, 25 (2024). https://doi.org/10.1007/s00203-023-03762-y
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DOI: https://doi.org/10.1007/s00203-023-03762-y