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Extremophiles

, Volume 22, Issue 1, pp 131–140 | Cite as

Biochemical characterization of a thermostable endomannanase/endoglucanase from Dictyoglomus turgidum

  • Francesca Anna Fusco
  • Raffaele Ronca
  • Gabriella Fiorentino
  • Emilia Pedone
  • Patrizia Contursi
  • Simonetta Bartolucci
  • Danila Limauro
Original Paper

Abstract

Dictyoglomus turgidum is a hyperthermophilic, anaerobic, gram-negative bacterium that shows an array of putative glycoside hydrolases (GHs) encoded by its genome, a feature that makes this microorganism very interesting for biotechnological applications. The aim of this work is the characterization of a hyperthermophilic GH5, Dtur_0671, of D. turgidum, annotated as endoglucanase and herein named DturCelB in agreement to DturCelA, which was previously characterized. The synthetic gene was expressed in Escherichia coli. The purified recombinant enzyme is active as a monomer (40 kDa) and CD structural studies showed a conserved α/β structure at different temperatures (25 and 70 °C) and high thermoresistance (Tm of 88 °C). Interestingly, the enzyme showed high endo-β-1,4-mannanase activity vs various mannans, but low endo-β-1,4 glucanase activity towards carboxymethylcellulose. The K M and V max of DturCelB were determined for both glucomannan and CMC: they were 4.70 mg/ml and 473.1 μmol/min mg and 1.83 mg/ml and 1.349 μmol/min mg, respectively. Its optimal activity towards temperature and pH resulted to be 70 °C and pH 5.4, respectively. Further characterization highlighted good thermal stability (~ 50% of enzymatic activity after 2 h at 70 °C) and pH stability over a broad range (> 90% of activity after 1 h in buffer, ranging pH 5–9); resistance to chemicals was also observed.

Keywords

Endomannanase Endoglucanase Thermophilic enzyme Dictyoglomus turgidum 

Notes

Acknowledgements

This work was supported by BIOPOLIS: PON03PE_00107_1 CUP: E48C14000030005.

Supplementary material

792_2017_983_MOESM1_ESM.pdf (723 kb)
Supplementary material 1 (PDF 722 kb)

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Copyright information

© Springer Japan KK, part of Springer Nature 2017

Authors and Affiliations

  • Francesca Anna Fusco
    • 1
  • Raffaele Ronca
    • 1
  • Gabriella Fiorentino
    • 1
  • Emilia Pedone
    • 2
  • Patrizia Contursi
    • 1
  • Simonetta Bartolucci
    • 1
  • Danila Limauro
    • 1
  1. 1.Dipartimento di BiologiaUniversity of Naples Federico II, Complesso universitario di Monte S. AngeloNaplesItaly
  2. 2.Istituto di Biostrutture e Bioimmagini, CNRNaplesItaly

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