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Computer-Assisted Rational Modifications to Improve the Thermostability of β-Glucosidase from Penicillium piceum H16

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Abstract

β-Glucosidase plays key role in the saccharification of cellulosic biomass. The use of β-glucosidase would benefit from improvements in its thermostability in some industrial applications. In this work, two rational computer-aided strategies were devised to enhance the thermostability of a highly active β-glucosidase from Penicillium piceum H16. Single-point mutants were proposed based on the proline theory. The effects of the mutation on protein stability were further predicted using molecular dynamics (MD) simulations. The optimal single mutant, G305P, was found to provide a 20.0 % improvement in thermostability. Then computer-assisted virtual saturated mutation was first used to select the optimal triple mutations which would further increase the thermostability of β-glucosidase by the Calculate Mutation Energy/Stability module of Discovery Studio (DS) 4.0. The triple mutant, S507F/Q512W/S514W, provided a 46.3 % improvement in thermostability. A detailed structural analysis showed that the additional hydrophobic interactions played decisive roles in the improvement in thermostability, and the use of mutants with enhanced stability may benefit formulation of more effective enzyme cocktails for lignocellulosic bioconversion.

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Acknowledgments

This work was supported by grants from the National High Technology Research and Development Program of China (863 Program) (No. 2012AA101807, 2014AA021906) and the Key Technology R&D program of Tianjin Municipal Science and Technology Commission (No. 13ZCZDSY05500), National Natural Science Foundation of China for the Youth (21406260).

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Authors and Affiliations

  1. Tianjin Key Laboratory for Industrial Biological Systems and Bioprocessing Engineering, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China

    Zhiyou Zong, Le Gao, Chao Cui, Shulin Chen & Dongyuan Zhang

  2. College of Chemistry, Nankai University, Tianjin, China

    Wensheng Cai

  3. Department of Biological Systems Engineering, Washington State University, Pullman, WA, 99164, USA

    Liang Yu

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  1. Zhiyou Zong
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  2. Le Gao
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  3. Wensheng Cai
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  4. Liang Yu
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  5. Chao Cui
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  6. Shulin Chen
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  7. Dongyuan Zhang
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Correspondence to Dongyuan Zhang.

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Zong, Z., Gao, L., Cai, W. et al. Computer-Assisted Rational Modifications to Improve the Thermostability of β-Glucosidase from Penicillium piceum H16. Bioenerg. Res. 8, 1384–1390 (2015). https://doi.org/10.1007/s12155-015-9603-4

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