Current Microbiology

, Volume 72, Issue 5, pp 583–588 | Cite as

Evaluation of Stress-Induced Microbial Siderophore from Pseudomonas aeruginosa Strain S1 as a Potential Matrix Metalloproteinase Inhibitor in Wound Healing Applications

  • Sita lakshmi Thyagarajan
  • S. Kandhasamy
  • Giriprasath Ramanathan
  • Uma Tiruchirapalli SivagnanamEmail author
  • P. T. PerumalEmail author


Matrix metalloproteinases (MMPs) are zinc-dependent proteolytic enzymes capable of causing various inflammatory and various degenerative diseases if over-expressed. The active site of these enzymes is a zinc binding motif which binds to the specific site on the substrate and induce degradation. Hence an inhibitor is required to form a complex with zinc motif which hampers the binding ability of MMPs. To obtain novel MMPs inhibitor for wound healing, the chelating activity of siderophore from the microbial source was focused. During screening for siderophore production, strain S1 produced the highest amount of siderophore in the minimal salts medium. The isolate was confirmed as Pseudomonas aeruginosa strain S1 based on 16S rRNA gene sequencing and phylogenetic analysis. The activity of the siderophore was assayed using chrome azurol sulphonate and purified by the chromatographic techniques. The structural evidence through Fourier transform infrared and nuclear magnetic resonance spectra revealed that the isolated siderophore is a catecholate type with the distinctive characters. The positive results of calcein and fluozin-3 assays indicate that siderophore could bind to divalent metal ions, namely Fe2+ and Zn2+. As the siderophore compound focused on wound healing property, the in vitro studies revealed the viability of NH3T3 fibroblast cells and its efficiency in matrix modulating was confirmed through gelatin zymogram.


Calcein Siderophore Production Minimal Salt Medium Metal Binding Ability Gelatin Zymogram 
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The author gratefully acknowledges financial support for this work through the grants awarded by the Department of Science and Technology New Delhi (SR/WOS-A/LS-375).

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

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Supplementary material 1 (DOC 1231 kb)


  1. 1.
    Albercht-Gary AM, Blane S, Rochel N, Ocaktan AZ, Abdallah MA (1994) Bacterial iron transport: coordination properties of pyoverdin PaA, a peptidic siderophore of Pseudomonas aeruginosa. Inorg Chem 33:6391–6402CrossRefGoogle Scholar
  2. 2.
    Altchul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSIBLAST: a new generation of protein database search programs. Nucleic Acids 25:3389–3402CrossRefGoogle Scholar
  3. 3.
    Auto Suomalainen K, Sorsa T (1990) Salivary collagenase: origin, characteristics and relationship to periodontal health. J Periodontal Res 25:135–142CrossRefGoogle Scholar
  4. 4.
    Baker AH, Edwards DR, Murphy G (2002) Metalloproteinase Inhibitors: biological actions and therapeutic opportunities. J Cell Sci 115:3719–3727CrossRefPubMedGoogle Scholar
  5. 5.
    Bode W, Maskos K (2001) Structural studies on MMPs and TIMPs. Methods Mol Biol 151:45–77PubMedGoogle Scholar
  6. 6.
    Bode W, Maskos K (2003) Structural basis of matrix metalloproteinases and their physiological inhibitors, the tissue inhibitors of metalloproteinases. Biol Chem 384:863–872CrossRefPubMedGoogle Scholar
  7. 7.
    Brosius J (1978) Complete nucleotide sequence of a 16S ribosomal gene of Escherichia coli. Proc Natl Acad Sci 75:4801–4805CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Brown PD (1998) Matrix metalloproteinase inhibitors. Breast Cancer Res Treat 52:125–136CrossRefPubMedGoogle Scholar
  9. 9.
    Drechsel H, Jung G (1998) Peptide siderophores. J Peptide Sci 4:147–181CrossRefGoogle Scholar
  10. 10.
    Gee KR, Zhou ZL, Qian WJ, Kennedy R (2002) Detection and imaging of zinc secretion from pancreatic β-cells using a new fluorescent zinc indicator. J Am Chem Soc 124:776–778CrossRefPubMedGoogle Scholar
  11. 11.
    Gendron R, Grenier D, Sorsa T, Uitto VJ, Mayrand D (1999) Effect of microbial siderophores on matrix metalloproteinase-2 activity. J Periodontal Res 34:50–53CrossRefPubMedGoogle Scholar
  12. 12.
    Haynes WE (1951) Pseudomonas aeruginosa—its characterization and identification. J Gen Microbiol 5:939–950CrossRefPubMedGoogle Scholar
  13. 13.
    Hujanen ES, Vaisanen A, Zheng A, Trygvason K, Turpeenniemi-Hujanen T (1994) Modulation of M (r) 72,000 and M (r) 92,000 type-IV collagenase (gelatinase A and B) gene expression by interferons alpha and gamma in human melanoma. Int J Cancer 58:582–586CrossRefPubMedGoogle Scholar
  14. 14.
    Itoh T, Tanioka M, Yoshida H, Nishimoto H, Itohara S (1998) Reduced angiogenesis and tumour progression in gelatinase A-deficient mice. Cancer Res 58:1048–1051PubMedGoogle Scholar
  15. 15.
    Meyer JM, Abdallah MA (1978) The fluorescent pigment Pseudomonas fluorescens, Biosynthesis, purification and physicochemical properties. J Gen Microbiol 107:319–328CrossRefGoogle Scholar
  16. 16.
    Neilands JB (1995) Siderophores: structure and function of microbial iron transport compounds. J Biol Chem 270:26723–26726CrossRefPubMedGoogle Scholar
  17. 17.
    O’Brien IG, Gibson F (1970) The structure of enterochelin and related 2,3-dihyroxy-N-benzouylserine conjugates from Escherichia coli. BioChem Biophys Acta 215:393–402CrossRefPubMedGoogle Scholar
  18. 18.
    Patel AK, Deshattiwar MK, Chaudhari BL, Chincholkar SB (2009) Production, purification and chemical characterization of the catecholate siderophore from potent probiotic strains of Bacillus sp. Bioresour Technol 100:368–373CrossRefPubMedGoogle Scholar
  19. 19.
    Payne SM (1994) Detection, isolation and characterization of siderophores. Method Enzymol 235:329–344CrossRefGoogle Scholar
  20. 20.
    Schwyn B, Neilands JB (1987) Universal chemical assay for the detection and determination of siderophores. Anal Biochem 160:47–56. doi: 10.1016/003-2697(87)90612-9 CrossRefPubMedGoogle Scholar
  21. 21.
    Scozzafava A, Supuran CT (2000) Carbonic anhydrase and matrix metalloproteinase inhibitors: sulfonylated amino acid hydroxmates with MMP inhibitory properties act as efficient inhibitors of CA isozymes I, II and IV and N-hydroxysulfonamides inhibit both these zinc enzymes. J Med Chem 43:3677–3687CrossRefPubMedGoogle Scholar
  22. 22.
    Sharman GJ, Williams DH, Ewing DF, Ratledge C (1995) Isolation, purification and tructure of exochelin MS, the extracellular siderophore from Mycobacterium smegmatis. J Biochem 305:187–196CrossRefGoogle Scholar
  23. 23.
    Sternlicht MD, Werb Z (2001) How matrix metalloproteinases regulate cell behavior. Annu Rev Cell Dev Biol 17:463–516CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Steward WP (1999) Marismastat (BB2516). Current status of development. Cancer Chemother Pharmacol 43:S56–S60CrossRefPubMedGoogle Scholar
  25. 25.
    Kandhasamy S, Ramanathan G, Kamalraja J, Balaji R, Mathivanan N, Sivagnanam UT, Perumal PT (2015) Synthesis, characterization and biological evaluation of chromen and pyrano chromen-5-one derivatives impregnated into a novel collagen based scaffold for tissue engineering applications. RSC Adv. doi: 10.1039/C5RA07133J Google Scholar
  26. 26.
    Tabraboletti G, Garofalo A, Belotti D, Drudis T, Borsotti P, Scanziani E, Brown PD, Giavazzi R (1995) Inhibition of angiogenesis and murine hemangioma growth by batismastat, a synthetic inhibitor of matrix metalloproteinases. J Natl Cancer Inst 87:293–298CrossRefGoogle Scholar
  27. 27.
    Utah Tara VJH, Huttunen A, Lindy S, Uitto J (1980) Activation of human leukocytes colonies by compounds reacting with sulfhydryl groups. Biochem Biophy Acta 613:168–177Google Scholar
  28. 28.
    Van Wart HE, Birkedal-Hansen H (1990) The cysteine switch: a principle of regulation of metalloproteinase activity with potential applicability to the entire matrix metalloproteinase gene family. Proc Natl Acad Sci 87:5578–5582CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Visca P (1993) Iron regulated salicylate synthesis by Pseudomonas sp. J Gen Microbiol 139:1995–2000CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Sita lakshmi Thyagarajan
    • 1
  • S. Kandhasamy
    • 2
  • Giriprasath Ramanathan
    • 1
  • Uma Tiruchirapalli Sivagnanam
    • 1
    Email author
  • P. T. Perumal
    • 2
    Email author
  1. 1.Bioproducts LabCSIR-Central Leather Research InstituteChennaiIndia
  2. 2.Organic DivisionCSIR-Central Leather Research InstituteChennaiIndia

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