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Purification and partial characterization οf a new β-xylosidase from Humicola grisea var. thermoidea

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Summary

The thermophilic fungus Humicola grisea var. thermoidea produces a mycelium-associated β-xylosidase activity when grown in liquid-state cultures on media containing oat spelt xylan as the carbon source. The β-xylosidase was purified to apparent homogeneity by gel filtration and anion exchange chromatography. Its molecular weight was 37 and 50 kDa, as determined by MALDI/TOF mass spectrometry and SDS-PAGE, respectively. The purified enzyme exhibited maximum activity at 55 °C and pH 6.5. It was also active at pH 8.8, retaining 60% of its activity after 6 h of incubation at 50 °C. β-xylosidase was strongly inactivated by NBS and slightly activated by DTT and β-mercaptoethanol. The enzyme was highly specific for PNPX as the substrate. The purified β-xylosidase showed K m and V max values of 1.37 mM and 12.98 IU  ml−1, respectively.

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References

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Acknowledgements

E.X.F.F. and T. I. acknowledge the receipt of research fellowship and post-graduate maintenance scholarship from CNPq (Brazil) and CAPES (Brazil), respectively.

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Correspondence to E.X.F. Filho.

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Iembo, T., Azevedo, M., Jr., C.B. et al. Purification and partial characterization οf a new β-xylosidase from Humicola grisea var. thermoidea . World J Microbiol Biotechnol 22, 475–479 (2006). https://doi.org/10.1007/s11274-005-9059-3

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Keywords

  • Characterization
  • chemical modification
  • Humicola grisea
  • purification
  • β-xylosidase