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SpyTag/SpyCatcher cyclization enhances the thermostability and organic solvent tolerance of l-phenylalanine aldolase

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Abstract

Objectives

To improve the thermostability and organic solvent tolerance of L-phenylserine aldolase, the in vivo SpyTag/SpyCatcher cyclization strategy was applied in this work.

Results

The in vivo cyclization of L-phenylserine aldolase was achieved by fusing the tags of SpyCatcher and SpyTag to the N- and C-termini of the enzyme, respectively. The kcat values and the circular dichroism spectra of the linear and cyclized LPAs are very similar, indicating that the cyclized LPA can be folded appropriately like the wild type. The cyclized enzyme has better thermostability and organic solvent tolerance than does the wild type. The half-life of L-phenylserine aldolase after cyclization was increased by 8.3 times at 70 °C, and the T50 also increased from 56.8 to 67.1 °C. The cyclized enzyme showed a remarkably higher tolerance to organic solvents (e.g., methanol, ethanol and acetone).

Conclusions

These results suggest that the in vivo cyclization using SpyTag/SpyCatcher is an effective strategy to improve the stability of enzymes, which potentially could be applied in industrial bioconversion.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (21476025). The authors have no conflicts of interest to declare.

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Author notes

  1. Yida Wang and Junwei Tian have contributed equally to this article.

Authors and Affiliations

  1. Department of Biological Science and Engineering, University of Science and Technology, Beijing, 100083, China

    Yida Wang, Ying Xiao, Hongxu Sun & Hui Luo

  2. Department of Environmental Engineering, University of Science and Technology, Beijing, 100083, China

    Junwei Tian, Yue Wang & Yanhong Chang

  3. Beijing Key Laboratory of Resource-oriented Treatment of Industrial Pollutants, Beijing, 100083, China

    Junwei Tian, Yue Wang & Yanhong Chang

Authors
  1. Yida Wang
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  2. Junwei Tian
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  3. Ying Xiao
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  4. Yue Wang
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  5. Hongxu Sun
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  6. Yanhong Chang
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  7. Hui Luo
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Corresponding authors

Correspondence to Yanhong Chang or Hui Luo.

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Wang, Y., Tian, J., Xiao, Y. et al. SpyTag/SpyCatcher cyclization enhances the thermostability and organic solvent tolerance of l-phenylalanine aldolase. Biotechnol Lett 41, 987–994 (2019). https://doi.org/10.1007/s10529-019-02689-z

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  • DOI: https://doi.org/10.1007/s10529-019-02689-z

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