Abstract
Staphylococcus aureus biofilms are the pathogenic factor in the spread of infection and are more pronounced in multidrug-resistant strains of S. aureus, where high expression of proteases is observed. Among various proteases, Serine protease (SspA) and cysteine protease Staphopain B (SspB) are known to play a key role in the biofilm formation and removal of biofilms. In earlier studies, we have reported Dibenzyl (benzo [d] thiazol-2-yl (hydroxy) methyl) phosphonate (DBTMP) exhibits anti-S. aureus and anti-biofilm properties by elevating the expression of the protease. In this study, the effect of DBTMP on the activities of SspA, and SspB of S. aureus was evaluated. The SspA and SspB genes of S. aureus ATCC12600 were sequenced (Genbank accession numbers: MZ456982 and MW574006). In S. aureus active SspA is formed by proteolytic cleavage of immature SspA, to get this mature SspA (mSspA), we have PCR amplified the mSspA sequence from the SspA gene. The mSspA and SspB genes were cloned, expressed, and characterized. The pure recombinant proteins rSspB and rmSspA exhibited a single band in SDS–PAGE with a molecular weight of 40 and 30 KD, respectively. The activities of rmSspA and rSspB are 32.33 and 35.45 Units/mL correspondingly. DBTMP elevated the activities of rmSspA and rSspB by docking with respective enzymes. This compound disrupted the biofilms formed by the multidrug-resistant strains of S. aureus and further prevented biofilm formation. These findings explain that DBTMP possesses anti-S. aureus and anti-biofilm features.
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Abbreviations
- SspA:
-
Staphylococcal serine protease A
- SspB:
-
Staphylococcal staphopain B
- DBTMP:
-
Dibenzyl (benzo [d] thiazol-2-yl (hydroxy) methyl) phosphonate
- mSspA:
-
Mature SspA
- ATCC:
-
American type cell culture
- PCR:
-
Polymerase chain reaction
- SDS–PAGE:
-
Sodium dodecyl sulfate–polyacrylamide gel electrophoresis
- KD:
-
Kilodalton
- EPS:
-
Extracellular polymeric substances
- MSCRAMMs:
-
Microbial surface components recognizing adhesive matrix molecules
- icaADBC:
-
Intercellular adhesion ADBC
- PIA:
-
Polysaccharide intercellular adhesin
- Fnbp:
-
Fibronectin-binding protein
- SpA:
-
Surface protein A
- Agr:
-
Accessory gene regulator
- SarA:
-
Staphylococcal accessory regulator
- LMV:
-
Local milk vendor
- D:
-
Dairy herds
- LB:
-
Luria–Bertani
- BHI:
-
Brain–Heart infusion
- TLC:
-
Thin layer chromatography
- SMA:
-
Skim milk agar
- CFU:
-
Colony forming units
- NCBI:
-
National Center for Biotechnology Information
- IPTG:
-
Isopropyl ß-d-1-thiogalactopyranoside
- MRSA:
-
Methicillin-resistant Staphylococcus aureus
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Acknowledgements
This study was supported by the Sri Venkateswara Institute of Medical Sciences and University, under the SBAVP scheme for providing funds and facilities to carry out this research work. This paper forms a part of my Ph.D. thesis work to be submitted to SVIMS University, Tirupati, Andhra Pradesh, India.
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This work was funded by Sri Balaji Arogya Vara Prasadini Scheme (SBAVP).
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PVGKS, AC, and GD: Designed the work. SSR and GD conducted the experiments, PVGKS, AC, and GD analyzed the results, and prepared and edited the manuscript. All the authors concurred with the manuscript.
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Deepika, G., Subbarayadu, S., Chaudhary, A. et al. Dibenzyl (benzo [d] thiazol-2-yl (hydroxy) methyl) phosphonate (DBTMP) showing anti-S. aureus and anti-biofilm properties by elevating activities of serine protease (SspA) and cysteine protease staphopain B (SspB). Arch Microbiol 204, 397 (2022). https://doi.org/10.1007/s00203-022-02974-y
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DOI: https://doi.org/10.1007/s00203-022-02974-y