Folia Microbiologica

, 46:315 | Cite as

Poly-3-hydroxybutyrate production byAzotobacter chroococcum

  • J. Parshad
  • S. Suneja
  • K. Kukreja
  • K. Lakshminarayana


Thirty-seven soil isolates and mutants ofAzotobacter chroococcum tested for poly-3-hydroxybutyrate (PHB) production using Sudan black B staining method were found to be positive. One mutant showed a higher number of PHB-producing cells and maximum number of granules per cell. Using 2% glucose and 15 mmol/L ammonium acetate, PHB production was found to be maximum at 36 and 48 h of growth under submerged cultivation and under stationary cultivation, respectively. PHB production was found to be higher on sucrose and commercial sugar (as carbon sources) as compared to glucose and mannitol. As commercial sugar is cheaper than sucrose it was selected as carbon source for PHB production, that being found to be maximum at 1% concentration. Inorganic nitrogen sources seemed to have no stimulatory effect on the production of PHB. However, ammonium acetate (15 mmol/L) was found to be best for PHB production. Peptone (0.2 %) gave a better yield of PHB under both growth conditions. Using all optimized conditions, PHB production was studied in ten selected strains. Two of them were found to be best PHB producers under both growth conditions, one producing 621 and 740 μg/g dry mass under submerged cultivation and under stationary cultivation, respectively, while the second one produced 589 and 733 μg/g.


Ammonium Acetate Inorganic Nitrogen Source Azotobacter Chroococcum Complex Nitrogen Source Peptone Concentration 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. Byrom I.: Polymer synthesis by microorganisms: technology and economics.Tibtech. 5, 246–250 (1987).Google Scholar
  2. Chen Q.G., Page W.J.: Production of poly-β-hydroxybutyrate byAzotobacter vinelandii in a two stage fermentation process.Biotechnol. Lett. 11, 347–350 (1997).Google Scholar
  3. Dawes E.A., Senior P.J.: The role and regulation of energy reserve polymers in microorganisms.Adv. Microbiol. Physiol. 10, 135–266 (1973).CrossRefGoogle Scholar
  4. Jensen V.: Notes on the biology ofAzotobacter.Proc. Soc. Appl. Bacteriol. 74, 89–93 (1951).Google Scholar
  5. Law J.H., Slepecky R.A.: Assay of poly-(β-hydroxybutyric acid.J. Bacteriol. 82, 33–36 (1961).PubMedGoogle Scholar
  6. Newton J.W., Wilson P.W., Burris R.H.: Direct demonstration of ammonia as an intermediate in nitrogen fixation byAzotobacter.J. Biol. Chem. 204, 445–451 (1953).PubMedGoogle Scholar
  7. Page W.J.: Production of poly-β-hydroxybutyrate byAzotobacter vinelandii UWD containing sugars and complex nitrogen sources.Appl. Microbiol. Biotechnol. 31, 329–333 (1992).Google Scholar
  8. Page W.J., Cornish A.: Growth ofAzotobacter vinelandii UWD in fish peptone medium and simplified extraction of poly-β-hydroxybutyrate.Appl. Environ. Microbiol. 59, 4236–4244 (1993).PubMedGoogle Scholar
  9. Page W.J., Knosp O.: Hyper-production of poly-β-hydroxybutyrate during exponential growth ofAzotobacter vinelandii UWD.Appl. Environ. Microbiol. 55, 1334–1339 (1989).PubMedGoogle Scholar
  10. Page W.J., Spherburne R., Elia L.D., Graham L.L.: Poly-β-hydroxybutyrate extrusion from pleomorphic cells ofAzotobacter vinelandii UWD.Can. J. Microbiol. 41, 22–31 (1995).CrossRefGoogle Scholar
  11. Pal S., Manna A., Paul A.K.: Nutritional and cultural conditions for production of poly-3-hydroxybutyric acid byAzotobacter chroococcum.Folia Microbiol. 43, 177–181 (1998).Google Scholar
  12. Schlegel H.G., Latterty R., Kraus I.: The isolation of mutants not accumulating poly-β-hydroxybutyric acid.Arch. Mikrobiol. 71, 283–294 (1970).PubMedCrossRefGoogle Scholar
  13. Senior P.J., Beech G.A., Ritchie G.A.F., Dawes E.A.: The role of oxygen limitation in the formation of poly-β-hydroxybutyrate during batch and continuous culture ofAzotobacter beijernckii.Biochem. J. 128, 1193–1201 (1972).PubMedGoogle Scholar
  14. Suguna Rani K.: Influence of plant growth promoting rhizobacteria on the population dynamics of the rhizosphere microflora, growth and development in cotton (Gossypium hirsutum).PhD Thesis. Tamil Nadu Agricultural University, Coimbatore (India) 2000.Google Scholar
  15. Thompson J.P., Skerman V.B.D.:Azotobacteriaceae: the Taxonomy and Ecology of the Aerobic Nitrogen-Fixing Bacteria. Academic Press, New York 1979.Google Scholar

Copyright information

© Folia Microbiologica 2001

Authors and Affiliations

  • J. Parshad
    • 1
  • S. Suneja
    • 1
  • K. Kukreja
    • 1
  • K. Lakshminarayana
    • 1
  1. 1.Department of MicrobiologyChaudhary Charan Singh Haryana Agricultural UniversityHisarIndia

Personalised recommendations