Antifungal and antibacterial activities of indigenous Streptomyces isolates from saline farmlands: prescreening, ribotyping and metabolic diversity

  • Imran Sajid
  • Clarisse Blandine Fotso Fondja Yao
  • Khaled Attia Shaaban
  • Shahida HasnainEmail author
  • Hartmut Laatsch
Original Paper


A culture collection of 110 indigenous Streptomyces strains originally isolated from saline farmlands (Punjab, Pakistan) using stringent methods was screened biologically and chemically to investigate their potential for the production of bioactive secondary metabolites. In a biological screening the crude extracts obtained from the culture broth of selected strains were analysed for their activity against a set of test organisms, including Gram-positive, Gram-negative bacteria, fungi and microalgae using the disk diffusion bioassay method. Additionally a cytotoxicity test was performed by means of the brine shrimp microwell cytotoxicity assay. In a chemical screening each of the crude extracts was analysed by TLC using various staining reagents and by HPLC-MS/MS measurements. The results depicted an impressive chemical diversity of crude extracts produced by these strains. The taxonomic status of the selected strains was confirmed by preliminary physiological testing and 16S rRNA gene sequencing.


Indigenous Streptomyces spp Prescreening Ribotyping Metabolite diversity HPLC-MS/MS 



We are thankful to Dr. H. Frauendorf for mass spectra, and F. Lissy and A. Kohl for technical assistance. A financial support of this work by a grant from Higher Education Commission (HEC) of Pakistan under IRSIP is gratefully acknowledged.


  1. Berdy J (2005) Bioactive microbial metabolites: a personal view. J Antibiot 58(1):1–26CrossRefGoogle Scholar
  2. Broach JR, Thorner J (1996) High throughput screening for drug discovery. Nature 384:14–16. doi: 10.1038/384014a0 CrossRefGoogle Scholar
  3. Burkhardt K, Fiedler HP, Grabley S, Thiericke R, Zeeck A (1996) New cineromycins and muscacins obtained by metabolic pattern analysis of Streptomyces griseoviridis (FH-S 1832) I. Taxonomy, fermentation, isolation, and biological activity. J Antibiot 49:432–437Google Scholar
  4. Edwards U, Rogall T, Bloecker H, Emde M, Boettger EC (1989) Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA. Nucleic Acids Res 17:7843–7853. doi: 10.1093/nar/17.19.7843 CrossRefGoogle Scholar
  5. Felnagle EA, Rondon MR, Berti AD, Crosby HA, Thomas MG (2007) Identification of the biosynthetic gene cluster and an additional gene for resistance to the antituberculosis drug capreomycin. Appl Environ Microbiol 73:4162–4170. doi: 10.1128/AEM.00485-07 CrossRefGoogle Scholar
  6. Fguira LF, Fotso S, Mehdi RA, Mellouli L, Laatsch H (2005) Purification and structure elucidation of antifungal and antibacterial activities of a newly isolated Streptomyces sp strain US80. Res Microbiol 156(3):341–347. doi: 10.1016/j.resmic.2004.10.006 CrossRefGoogle Scholar
  7. Gerhardt P, Murry RGE, Wood WA, Kreig NR (1994) Methods for general and molecular bacteriology. ASM Washington, DCGoogle Scholar
  8. Grabley S, Thiericke R, Zeeck A (1999) The chemical screening approach. In: Grabley S, Thiericke R (eds) Drug discovery from nature. Springer–Verlag, Heidelberg, pp 124–148Google Scholar
  9. Hayakawa M, Yoshida Y, Iimura Y (2004) Selective isolation of bioactive soil actinomycetes belonging to the Streptomyces violaceusniger phenotypic cluster. J Appl Microbiol 96:973–981. doi: 10.1111/j.1365-2672.2004.02230.x CrossRefGoogle Scholar
  10. Kantz T, Bold HC (1969) Morphological and taxonomic investigation of Nostoc Anabena in culture. Physiological studies, University of Texas (publication No. 6924), Austin, Texas, USAGoogle Scholar
  11. Laatsch H (2000) Mikroorganismen als biologische Quelle neuer Wirkstoffe. In: Kayser O, Müller RH (eds) Pharmazeutische Biotechnologie. Wissenschaftliche Verlagsges, Stuttgart, Germany, pp 13–43Google Scholar
  12. Laatsch H (2007) AntiBase 2007, a data base for rapid dereplication and structure determination of microbial natural products. Wiley–VCH: Weinheim, Germany (and annual updates); see http://wwwuser.gwdgde/~ucoc/laatsch/AntiBase.htm
  13. Lazzarini A, Cavaletti L, Toppo G, Marinelli F (2001) Rare genera of actinomycetes as potential sources of new antibiotics. Antonie Van Leeuwenhoek 78:399–405. doi: 10.1023/A:1010287600557 CrossRefGoogle Scholar
  14. Lechevalier HA, Williams ST, Sharpe ME, Holt JG (1989) The Actinomycetes: a practical guide to genetic identification of actinomycetes. In: Bergey’s manual of systematic bacteriology. pp 2344–3330Google Scholar
  15. Locci R (1989) Streptomycetes and related genera. In: Williams ST, Sharpe ME, Holt JG (eds) Bergey’s manual of systematic bacteriology. Williams & Wilkins, Baltimore, pp 2451–2493Google Scholar
  16. Omura S (1992) The search for bioactive compounds from microorganisms, 1st edn. Springer, Berlin, Heidelberg, New YorkGoogle Scholar
  17. Solis PN, Wright CW, Anderson MM, Gupta MP, Phillipson JD (1993) A microwell cytotoxicity assay using Artemia salina (brine shrimp). Planta Med 59:250–252. doi: 10.1055/s-2006-959661 CrossRefGoogle Scholar
  18. Taddei A, Valderrama M, Giarrizzo J, Rey M, Castelli C (2006) Chemical screening: a simple approach to visualizing Streptomyces diversity for drug discovery and further research. Res Microbiol 157:291–297. doi: 10.1016/j.resmic.2005.07.007 CrossRefGoogle Scholar
  19. Takahashi Y, Omura S (2003) Isolation of new actinomycete strains for the screening of new bioactive compounds. J Gen Appl Microbiol 49:141–154. doi: 10.2323/jgam.49.141 CrossRefGoogle Scholar
  20. Watve MG, Tickoo R, Jog MM, Bhole BD (2001) How many antibiotics are produced by the genus Streptomyces? Arch Microbiol 157:386–390. doi: 10.1007/s002030100345 CrossRefGoogle Scholar
  21. Wendisch FK, Kutzner HJ (1991) The family Streptomycetaceae. In: Balows A, Trüper HG, Dworkin M, Harder W, Schleifer KH (eds), The prokaryotes, A handbook on the biology of bacteria: ecophysiology, isolation, identification, applications. Springer–Verlag, Berlin/New York, pp 965–968Google Scholar
  22. Yun BS, Hidaka T, Furihata K, Seto H (1994) Microbial metabolites with tip A promoter inducing activity. J Antibiot 47:969–975Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Imran Sajid
    • 1
    • 2
  • Clarisse Blandine Fotso Fondja Yao
    • 2
  • Khaled Attia Shaaban
    • 2
  • Shahida Hasnain
    • 1
    Email author
  • Hartmut Laatsch
    • 2
  1. 1.Department of Microbiology and Molecular GeneticsUniversity of the PunjabLahorePakistan
  2. 2.Institute of Organic and Biomolecular Chemistry, University of GöttingenGottingenGermany

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