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Rational protein design for thermostabilization of glycoside hydrolases based on structural analysis

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Abstract

Glycosidases are used in the food, chemical, and energy industries. These proteins are some of the most frequently used such enzymes, and their thermostability is essential for long-term and/or repeated use. In addition to thermostability, modification of the substrate selectivity and improvement of the glycosidase activities are also important. Thermostabilization of enzymes can be performed by directed evolution via random mutagenesis or by rational design via site-directed mutagenesis; each approach has advantages and disadvantages. In this paper, we introduce thermostabilization of glycoside hydrolases by rational protein design using site-directed mutagenesis along with X-ray crystallography and simulation modeling. We focus on the methods of thermostabilization of glycoside hydrolases by linking the N- and C-terminal ends, introducing disulfide bridges, and optimizing β-turn structures to promote hydrophobic interactions.

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Funding

This study was supported by JSPS KAKENHI (Grant-in-Aid for Young Scientists B, Grant No. 26850067).

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Author notes

  1. Masahiro Watanabe and Tomohiko Matsuzawa contributed equally to this work.

Authors and Affiliations

  1. Research Institute for Sustainable Chemistry, National Institute of Advanced Industrial Science and Technology (AIST), 3-11-32, Kagamiyama, Higashihiroshima, Hiroshima, 739-0046, Japan

    Masahiro Watanabe

  2. Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan

    Tomohiko Matsuzawa & Katsuro Yaoi

Authors
  1. Masahiro Watanabe
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  2. Tomohiko Matsuzawa
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  3. Katsuro Yaoi
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Correspondence to Katsuro Yaoi.

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Watanabe, M., Matsuzawa, T. & Yaoi, K. Rational protein design for thermostabilization of glycoside hydrolases based on structural analysis. Appl Microbiol Biotechnol 102, 8677–8684 (2018). https://doi.org/10.1007/s00253-018-9288-7

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  • DOI: https://doi.org/10.1007/s00253-018-9288-7

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