Journal of Polymers and the Environment

, Volume 18, Issue 2, pp 141–147 | Cite as

Purification and Properties of Extracellular Poly(3-hydroxybutyrate) Depolymerase Produced by Aspergillus fumigatus 202

  • Rachana Bhatt
  • K. C. Patel
  • Ujjval Trivedi
Original Paper


An extracellular poly(3-hydroxybutyrate) (PHB) depolymerase produced by a thermotolerant fungal soil isolate, Aspergillus fumigatus 202, was purified and characterized. Maximum PHB depolymerase production was obtained at the end of 48 h with initial medium pH 7.0 and 45 °C in Bushnell Haas Minerals medium containing PHB as sole source of carbon. The PHB depolymerase was purified using size exclusion chromatography to a fold purification of 20.62 and 61.62% yield. SDS-PAGE and isoelectric focusing revealed the molecular weight and pI of the purified enzyme as 63,744 Da and 4.2, respectively. N-terminal amino acid sequence of purified enzyme was HAXDAYLVK. This non-glycosylated enzyme was most active at pH 9.0 and 45 °C. Purified enzyme was inactivated by N-bromosuccinimide and dithiothreitol suggesting the involvement of tryptophan residues and disulfide bonds at its active site. Nonionic detergents like Tween 20, Tween 80 and Triton X-100 inhibited the enzyme activity. Ions like Ca+2 and Mg+2 (5 mM) increased the enzyme activity 1.5 times. Fe+2 effectively inhibited the enzyme activity to 88% whereas Hg+2 completely inhibited the enzyme.


Poly(3-hydroxybutyrate) PHB depolymerase Aspergillus fumigatus 202 Thermotolerant fungus Purification 


  1. 1.
    Hazer B, Steinbuchel A (2007) Appl Microbiol Biotechnol 74:1–12CrossRefGoogle Scholar
  2. 2.
    Bhatt R, Shah D, Patel K, Trivedi U (2008) Biores Technol 99:4615–4620CrossRefGoogle Scholar
  3. 3.
    Asrar J, Gruys K (2002) In Doi Y, Steinbuchel A (eds) Biopolymers, vol 4. Wiley-VCH, Weinheim, pp 53–84Google Scholar
  4. 4.
    Jendrossek D, Handrick R (2002) Annu Rev Microbiol 56:403–432CrossRefGoogle Scholar
  5. 5.
    Brucato C, Wong S (1991) Arch Biochem Biophys 290:497–502CrossRefGoogle Scholar
  6. 6.
    Iyer S, Shah R, Sharma A, Jendrossek D, Desai A (2000) J Polym Environ 8:197–203CrossRefGoogle Scholar
  7. 7.
    Oda Y, Osaka H, Urakami T, Tonomura K (1997) Curr Microbiol 34:230–232CrossRefGoogle Scholar
  8. 8.
    Liu H, Zhang H, Chen S, Liu D, Xia H (2006) J Polym Environ 14:419–426CrossRefGoogle Scholar
  9. 9.
    Jendrossek D, Muller B, Schelgel H (1993) Eur J Biochem 218:701–710CrossRefGoogle Scholar
  10. 10.
    Kita K, Ishimaru K, Teraoka M, Yanase H, Kato N (1995) Appl Environ Microbiol 61:1727–1730Google Scholar
  11. 11.
    Laemmli U (1970) Nature 227:680–685CrossRefGoogle Scholar
  12. 12.
    Doerner K, White B (1990) Anal Biochem 187:147–150CrossRefGoogle Scholar
  13. 13.
    Mergaert J, Wouters A, Anderson C, Swing J (1995) Can J Microbiol 41:154–159CrossRefGoogle Scholar
  14. 14.
    Han J, Kim M (2002) J Microbiol 40:20–25Google Scholar
  15. 15.
    Han J, Son Y, Chang C, Kim M (1998) J Microbiol 36:67–73Google Scholar
  16. 16.
    Kim D, Yun J, Kim H, Bae K, Rhee Y (2002) J Microbiol 40:129–133Google Scholar
  17. 17.
    Scherer T, Fuller R, Lenz R, Goodwin S (1999) J Environ Polym Degrad 7:117–125CrossRefGoogle Scholar
  18. 18.
    Su-Qin C, Shan C, Liu D, Xia H (2006) World J Microbiol Biotechnol 22:729–735CrossRefGoogle Scholar
  19. 19.
    Sadocco P, Nacerino S, Paglia E, Seves A, Elegir G (1997) J Environ Polym Degrad 5:57–65Google Scholar
  20. 20.
    Kasuya K, Inoune Y, Tanaka T, Akehata T, Iwata T (1997) Appl Environ Microbiol 63:4844–4852Google Scholar
  21. 21.
    Spande T, Green N, Witkop B (1966) Biochemistry 5:1926–1933CrossRefGoogle Scholar
  22. 22.
    Matovulj M, Molitoris H (1992) FEMS Microbiol Rev 103:323–332CrossRefGoogle Scholar
  23. 23.
    Mergaert J, Webb A, Anderson C, Wouters A, Swing J (1993) Appl Environ Microbiol 59:3233–3238Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.BRD School of BiosciencesSardar Patel UniversityVallabh VidyanagarIndia

Personalised recommendations