Isolation of Streptomyces Species from Soil

Madan K. Kharel1, Micah D. Shepherd1, Stephen E. Nybo1, Michael L. Smith1, Mary A. Bosserman1, Jürgen Rohr1

1 Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, Kentucky
Publication Name:  Current Protocols in Microbiology
Unit Number:  Unit 10E.4
DOI:  10.1002/9780471729259.mc10e04s19
Online Posting Date:  November, 2010
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Abstract

This unit describes a general protocol for the isolation of Streptomyces species from soil and fresh water, using a procedure for the selective growth of Streptomyces species. Preparation of the necessary growth medium, recognition of the morphology of the bacteria, and safety considerations are also covered. Curr. Protoc. Microbiol. 19:10E.4.1‐10E.4.5. © 2010 by John Wiley & Sons, Inc.

Keywords: Streptomyces; antibiotics; isolation

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Isolation of Streptomyces Species from Soil and from Fresh Water
  • Support Protocol 1: Preparation of Solid Agar Medium for Isolation of Streptomyces
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Isolation of Streptomyces Species from Soil and from Fresh Water

  Materials
  • Soil
  • Sterile solid media (M2 agar, oatmeal agar, and ISP4 agar plates) supplemented with nalidixic acid and cycloheximide ( protocol 1)
  • 50‐ml tubes, sterile
  • 1.5‐ml microcentrifuge tubes, sterile
  • Vortex
  • Water bath
  • Biological safety cabinet
  • Sterile spreader
  • 28°C incubator
  • Sterile tooth picks
  • Microscope slides
  • Scanning election microscope, optional

Support Protocol 1: Preparation of Solid Agar Medium for Isolation of Streptomyces

  Materials
  • Cycloheximide
  • Nalidixic acid
  • NaOH solution
  • M2 agar, oatmeal agar, or ISP4 agar mixtures (see reciperecipes)
  • 15‐ml sterile tubes (used for preparing the antibiotic solutions)
  • 0.25‐µm syringe filter, sterile
  • Stirring magnetic bar
  • Autoclave
  • 100 × 15–mm Petri dishes
  • Parafilm
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Figures

Videos

Literature Cited

Literature Cited
   Bérdy, J. 1995. Are Actinomycetes exhausted as a source of secondary metabolites? Proc. 9th Symp. Actinomycetes pp. 13‐34.
   Miyadoh, S. 1993. Research on antibiotic screening in Japan over the last decade: A producing microorganisms approach. Actinomycetologica 9:100‐106.
   Nonomura, H. 1974. Key for classification and identification of 458 species of the Streptomycetes included in ISP. J. Ferment. Technol. 52:78‐92.
   Sanglier, J.J., Haag, H., Huck, T.A., and Fehr, T. 1993. Novel bioactive compounds from actinomycetes—a short review (1988‐1992). Res. Microbiol. 144:633‐642.
   Shirling, E.B. and Gottlieb, D. 1966. Method for characterizing Streptomyces species. Int. J. Syst. Bacteriol. 16:313‐340.
   Vining, L.C. 1990. Functions of secondary metabolites. Ann. Rev. Microbiol. 44:395‐427.
   Watve, M.G., Tickoo, R., Jog, M.M., and Bhole, B.D. 2001. How many antibiotics are produced by the genus Streptomyces? Arch. Microbiol. 176:386‐390.
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