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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

Abstract

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.

Keywords

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

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.BRD School of BiosciencesSardar Patel UniversityVallabh VidyanagarIndia

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