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Unusual hydrophobic linker region of β-glucosidase (BGLII) from Thermoascus aurantiacus is required for hyper-activation by organic solvents

Abstract

A gene encoding a putative β-glucosidase was isolated from Thermoascus aurantiacus IFO9748 and designated as bgl2. The recombinant enzyme showed β-glucosidase activity when p-nitrophenyl-β-glucose (pNP-Glc) was used as substrate. We also found that the enzyme activity was increased in the presence of organic solvents. An addition of 20 % (v/v) 1-octanol resulted in 54-fold higher activity of pNP-Glc hydrolysis, and transglycosylation activity was also found to be activated. The results of tryptophan fluorescence spectral analysis revealed the changes in the tertiary structure of the enzyme in the presence of 1-hexanol that may cause increased enzyme activity. BGLII has a distinctive hydrophobic linker region between N- and C-terminal domains. A chimeric enzyme in which the linker region was substituted by the corresponding region of another β-glucosidase failed to be activated by organic solvents, suggesting that the hydrophobic linker region may act as a molecular switch in BGLII.

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Acknowledgements

We thank Kenji Yamamoto, Graduate School of Biostudies, Bunzo Mikami and Noboyuki Takahashi, Graduate School of Agriculture, Kyoto University, for their invaluable suggestions. This work was supported by grants-in-aid for scientific research from the Ministry of Education, Science, Sports and Culture of Japan, the New Energy and Industrial Technology Development Organization, Japan Society for the Promotion of Science, and the Research Institute of Innovative Technology for the Earth.

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Correspondence to Hisanori Tamaki.

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Hong, J., Tamaki, H. & Kumagai, H. Unusual hydrophobic linker region of β-glucosidase (BGLII) from Thermoascus aurantiacus is required for hyper-activation by organic solvents. Appl Microbiol Biotechnol 73, 80–88 (2006). https://doi.org/10.1007/s00253-006-0428-0

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Keywords

  • Cellobiose
  • Recombinant Enzyme
  • Tryptophan Fluorescence
  • Transglycosylation Activity
  • Transglycosylation Reaction