Acta Biologica Hungarica

, Volume 59, Issue 4, pp 489–499 | Cite as

A Novel Poly(3-Hydroxybutyrate)-Degrading Streptoverticillium Kashmirense AF1 Isolated from Soil and Purification of Phb-Depolymerase

  • A. A. ShahEmail author
  • Fariha Hasan
  • A. Hameed
  • Safia Ahmed


A new bacterial strain, capable of degrading poly(3-hydroxybutyrate) (PHB) was isolated from soil. This organism, identified as Streptoverticillium kashmirense AF1, secreted PHB depolymerases both on solid as well as in liquid mineral salt medium containing poly(3-hydroxybutyrate) as sole carbon source. The optimum production of PHB depolymerase was observed at pH 8 and 7, at 45 °C, 1% substrate concentration and in the presence of lactose as an additional carbon source. The extracellular PHB depolymerase was purified by gel permeation chromatography using Sephadex G-75. The Streptoverticillium kashmirense AF1 produced two types of PHB depolymerases having molecular weights of about 37 and 45 kDa as determined by SDS-PAGE. The difference in dry cell mass and amount of CO2 evolved in the test and control calculated gravimetrically through Sturm test indicated the degradative capabilities of Streptoverticillium kashmirense AF1.


Biodegradation poly(3-hydroxybutyrate) poly(3-hydroxybutyrate) depolymerase Streptoverticillium kashmirense AF1 purification 


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  1. 1.
    Abou-Zeid, D. M., Müller, R. J., Deckwer, W. D. (2001) Degradation of natural and synthetic polyesters under anaerobic conditions. J. Biotechnol. 86, 113–126.CrossRefGoogle Scholar
  2. 2.
    Briese, B. H., Schmidt, B., Jendrossek, D. (1994) Pseudomonas lemoignei has five poly(hydroxy-alkanoic acid) (PHA) depolymerase genes: a comparative study of bacterial and eukaryotic PHA depolymerases. J. Environ. Polym. Degrad. 2, 75–87.CrossRefGoogle Scholar
  3. 3.
    Calabia, B. P., Tokiwa, Y. (2006) A novel PHB depolymerase from a thermophilic Streptomyces sp. Biotechnol. Lett. 28, 383–388.CrossRefGoogle Scholar
  4. 4.
    Ghanem, N. B., Mabrouk, M. E., Sabry, S. A., El-Badan, D. E. (2005) Degradation of polyesters by a novel marine Nocardiopsis aegyptia sp. nov.: application of Plackett-Burman experimental design for the improvement of PHB depolymerase activity. J. Gen. Appl. Microbiol. 51, 151–158.CrossRefGoogle Scholar
  5. 5.
    Holt, G. (1993) Bergey’s Manual of Determinative Bacteriology, 9th ed., The Williams and Wilkins Company, Baltimore.Google Scholar
  6. 6.
    Imam, S. H., Gordon, S. H., Shogren, R. L., Tosteson, T. R., Govind, N. S., Greene, R. V. (1999) Degradation of starch-poly(β-hydroxybutyrate-co-β-hydroxyvalerate) bioplastic in tropical coastal waters. Appl. Environ. Microbiol. 65, 431–437.PubMedPubMedCentralGoogle Scholar
  7. 7.
    Kim, M. N., Lee, B. Y., Lee, I. M., Lee, H. S., Yoon, J. S. (2001) Toxicity and biodegradation of products from polyester hydrolysis. J. Environ. Sci. Health A. Tox. Hazard. Subst. Environ. Eng. 36, 447–463.CrossRefGoogle Scholar
  8. 8.
    Kobayashi, T, Sugiyama, A., Kawase, Y, Saito, T, Mergaert, J., Swings, J. (1999) Biochemical and genetic characterization of an extracellular poly(3-hydroxybutyrate) depolymerase from Acidovorax sp. strain TP4. J. Environ. Polym. Degrad. 7, 9–17.CrossRefGoogle Scholar
  9. 9.
    Manna, A., Paul, A. K. (2000) Degradation of microbial polyester poly(3-hydroxybutyrate) in environmental samples and in culture. Biodegradation 11, 323–329.CrossRefGoogle Scholar
  10. 10.
    Manna, A., Giri, P., Paul, A. K. (1999) Degradation of poly(3-hydroxybutyrate) by soil strepto-mycetes. World J. Microbiol. Biotechnol. 15, 705–709.CrossRefGoogle Scholar
  11. 11.
    Mergaert, J., Webb, A., Anderson, C., Wouters, A., Swings, J. (1993) Microbial degradation of poly(3-hydroxybutyrate) and poly(3-hydroxybutyrateco-3-hydroxyvalerate) in soils. Appl. Environ. Microbiol. 59, 3233–3238.PubMedPubMedCentralGoogle Scholar
  12. 12.
    Mergaert, J., Wouters, A., Swings, J. (1994) Estimation of the intrinsic biodiversity among poly(3-hydroxyalkanoates) degrading streptomycetes using gas chromatographic analysis of fatty acids. Syst. Appl. Microbiol. 17, 601–612.CrossRefGoogle Scholar
  13. 13.
    Mergaert, J., Wouters, A., Anderson, C., Swings, J. (1995) In situ biodegradation of poly(3-hydroxy-butyrate) and poly(3- hydroxybutyrate-co-3-hydroxyvalerate) in natural waters. Can. J. Microbiol. 41, 154–159.CrossRefGoogle Scholar
  14. 14.
    Mergaert, J., Schirmer, A., Hauben, L., Mau, M., Hoste, B., Kerstens, K., Jendrossek, D., Swings, J. (1996) Isolation and identification of poly(3-hydroxy valerate)-degrading strains of Pseudomonas lemoignei. Int. J. Syst. Bacteriol. 46, 769–773.CrossRefGoogle Scholar
  15. 15.
    Mukai, K., Doi, Y., Sema, Y., Tomita, K. (1993) Substrate specificities in hydrolysis of polyhydroxy-alkanoates by microbial esterases. Biotechnol. Lett. 15, 601–604.CrossRefGoogle Scholar
  16. 16.
    Muller, R. J., Augusta, J., Pantke, M. (1992) An Interlaboratory investigation into biodegradation of plastics; Part 1: A modified Sturm-test. Mat. Organ. 27, 179–189.Google Scholar
  17. 17.
    Nishida, H., Tokiwa, Y (1993) Distribution of poly(P-hydroxybutyrate) and poly(e-caprolactone) aerobic degrading microorganisms in different environments. J. Environ. Polym. Degrad. 1, 227–233.CrossRefGoogle Scholar
  18. 18.
    Nojiri, M., Saito, T. (1997) Structure and function of poly(3-hydroxybutyrate) depolymerase from Alcaligenes faecalis Tl. J. Bacteriol. 179, 6965–6970.CrossRefGoogle Scholar
  19. 19.
    Pagga, U., Beimborn, D. B., Boelens, J., De Wilde, B. (1995) Determination of the aerobic biodegradability of polymeric material in a laboratory controlled composting test. Chemosphere 31, 4475–4487.CrossRefGoogle Scholar
  20. 20.
    Pagga, U., Schafer, A., Muller, R. J., Pantke, M. (2001) Determination of the aerobic biodegradability of polymeric material in aquatic batch tests. Chemosphere 42, 319–331.CrossRefGoogle Scholar
  21. 21.
    Reinhardt, S., Handrick, R., Jendrossek, D. (2002) The “PHB depolymerase inhibitor” of Pauci-monas lemoignei is a PHB depolymerase. Biomacromolecule 3, 823–827.CrossRefGoogle Scholar
  22. 22.
    Romen, F., Reinhardt, S., Jendrossek, D. (2004) Thermotolerant poly(3-hydroxybutyrate)-degrading bacteria from hot compost and characterization of the PHB depolymerase of Schlegelella sp. KBla. Arch. Microbiol. 182: 157–164.CrossRefGoogle Scholar
  23. 23.
    Schirmer, A., Jendrossek, D., Schlegel, H. G. (1993) Degradation of poly(3-hydroxyoctanoic acid) [P(3HO)] by bacteria: purification and properties of a P(3HO) depolymerase from Pseudomonas fluorescens GK13. Appl. Environ. Microbiol. 59, 1220–1227.PubMedPubMedCentralGoogle Scholar
  24. 24.
    Shah, A. A., Hasan, F., Hameed, A., Ahmed, S. (2007) Isolation and characterization of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) degrading bacteria and purification of PHBV depolymerase from newly isolated Bacillus sp. AF3. Int. Biodeterior Biodegrad. 60, 109–115.CrossRefGoogle Scholar
  25. 25.
    Tansengco, M. L., Tokiwa, Y (1998) Thermophilic microbial degradation of polyethylene succinate. World J. Microbiol. Biotechnol. 14, 133–138.CrossRefGoogle Scholar
  26. 26.
    Tokiwa, Y., Calabia, B. P. (2004) Degradation of microbial polyesters. Biotechnol. Lett. 26, 1181–1189.CrossRefGoogle Scholar
  27. 27.
    Whitchurch, G. J., Cooper, T. A. (2006) Thermoplastically-processable biodegradable polyvinyl alcohol biodegradable polyvinyl alcohol-, starch starch- and polylactide and polylactide-based polymer blends. International Degradable Plastics Symposium Chicago IL, USA, 2006.Google Scholar
  28. 28.
    Yamada, K., Mukai, K., Doi, Y. (1993) Enzymatic degradation of poly-(hydroxy-alkanoates) by Pseudomonas pickettii. Int. J. Biol. Macromol. 15, 215–220.CrossRefGoogle Scholar

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© Akadémiai Kiadó, Budapest 2008

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (, which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Authors and Affiliations

  • A. A. Shah
    • 1
    Email author
  • Fariha Hasan
    • 1
  • A. Hameed
    • 1
  • Safia Ahmed
    • 1
  1. 1.Department of MicrobiologyQuaid-i-Azam UniversityIslamabadPakistan

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