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Biochemical characterization of a thermostable endomannanase/endoglucanase from Dictyoglomus turgidum

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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.

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Acknowledgements

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

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Correspondence to Danila Limauro.

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Communicated by H. Atomi.

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Fusco, F.A., Ronca, R., Fiorentino, G. et al. Biochemical characterization of a thermostable endomannanase/endoglucanase from Dictyoglomus turgidum . Extremophiles 22, 131–140 (2018). https://doi.org/10.1007/s00792-017-0983-6

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  • DOI: https://doi.org/10.1007/s00792-017-0983-6

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