Biotechnology Techniques

, Volume 8, Issue 5, pp 345–350 | Cite as

Staining method of poly(3-hydroxyalkanoic acids) producing bacteria by nile blue

  • Shiro Kitamura
  • Yoshihara Doi


Using an ethanol solution of nile blue, we have developed an efficient method to detect the colonies of poly(3-hydroxyalkanoic acids) (PHA) producing bacteria on the agar plate. When the bacterial colonies with PHA granules were stained with nile blue, the stained colonies fluoresced bright orange on the irradiation of UV light. In the fluoresce emission spectra, fluorescence intensity increased with an increase in the PHA content of bacterial cells.Alcaligenes eutrophus andA.latus colonies with poly(3-hydroxybutyric acid) (PHB) homopolymer exhibited an emission maximum at 580nm on the excitation at 490nm. On the other hand,Pseudomonas oleovorans andP.putida with medium-chain-length (mcl-) PHA copolymers of C6, C8 and C10 units exhibited an emission maximum at 570nm.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Anderson, A. J. and Dawes, E.A. (1990).Microbiol. Rev., 54, 450–472.Google Scholar
  2. Doi, Y. (1990). Microbial polyesters, VHC, New York.Google Scholar
  3. Doi, Y., Tamaki, A., Kunioka, M., and Soga, K. (1987).J. Chem. Soc., Chem. Commun., 1635–1636.Google Scholar
  4. Haywood, G.W., Anderson, A.J., Ewing, D.F., and Dawes, E.A. (1990).Appl. Environ. Microbiol., 56, 3354–3359.Google Scholar
  5. Lageveen, R.G., Huisman, G.W., Preusting, H., Ketelaar, P., Eggink, G., and Witholt, B. (1988).Appl. Environ. Microbiol., 54, 2924–2932.Google Scholar
  6. Ostle, A. G. and Holt, J. G. (1982).Appl. Environ. Microbiol., 44, 238–241.Google Scholar
  7. Ramsay, B.A., Lomaliza, K., Chavarie, C., Dube, B., Bataille, P., and Ramsay, J.A. (1990).Appl Environ. Microbiol., 56, 2093–2098.Google Scholar
  8. Repaske, R. and Repaske, A. C. (1976).Appl. Environ. Microbiol., 32,. 585–591.Google Scholar
  9. Schlegel, H.G., Lafferty, R., and Krauss, I. (1970).Arch. Microbiol., 71, 283–294.Google Scholar
  10. Steinbüchel, A. and Schlegel, H.G. (1989).Appl. Microbiol. Biotechnol., 31, 168–175.CrossRefGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1994

Authors and Affiliations

  • Shiro Kitamura
    • 1
  • Yoshihara Doi
    • 2
  1. 1.Research Institute of Innovative Technology for the Earth (RITE)SaitamaJapan
  2. 2.Polymer Chemistry Laboratory, The Institute of Physical and Chemical Research (RIKEN)SaitamaJapan

Personalised recommendations