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The Stability Enhancement of Nitrile Hydratase from Bordetella petrii by Swapping the C-terminal Domain of β subunit

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Abstract

The thermal stability of most nitrile hydratases (NHase) is poor, which has been enhanced to some extent by molecular modifications in several specific regions of the C-terminal domain (C-domain) of β subunit of NHase. Since the C-domain could be present as a naturally separate domain in a few NHases, the whole C-domain is proposed to be related to the NHase stability. The chimeric NHase (SBpNHase) from the thermal-sensitive BpNHase (NHase from Bordetella petrii) and the relatively thermal-stable PtNHase (NHase from Pseudonocardia thermophila) was constructed by swapping the corresponding C-domains. After 30 min incubation at 50 °C, the original BpNHase nearly lost its activity, while the SBpNHase retained 50 % residual activity, compared with the melting temperature (Tm) (50 °C) of the original BpNHase, that of the SBpNHase was 55 °C. The SBpNHase with higher thermal stability would be useful for the thermal stability enhancement of NHase and for the understanding of the relationship between the stability of NHase and its structure.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (21506172), the Spring Sunshine Plan of the Ministry of Education of China (Z2015121), the Key Project of Xihua University of China (z1420524) and (z1420525), the Department of Education of Sichuan Province of China (15205445), and the Key Laboratory program of Food and Biotechnology of Xihua University of China (szjj2015-013) and (szjj2015-014).

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    Authors and Affiliations

    1. Key Laboratory of Food and Biotechnology, School of Food and Biotechnology, Xihua University, Chengdu, Sichuan, 610039, People’s Republic of China

      Weifeng Sun, Xianggui Chen, Lunjie Wu & Yi Liu

    2. School of Biochemical Engineering, Anhui Polytechnic University, Anhui, 241000, China

      Longbao Zhu

    3. Key Laboratory of Industrial Biotechnology, School of Biotechnology, Jiangnan University, Wuxi, 214122, China

      Zhemin Zhou

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    1. Weifeng Sun
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    2. Longbao Zhu
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    3. Xianggui Chen
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    4. Lunjie Wu
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    5. Zhemin Zhou
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    6. Yi Liu
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    Corresponding author

    Correspondence to Yi Liu.

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    Weifeng Sun and Longbao Zhu contributed equally to this work.

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    Sun, W., Zhu, L., Chen, X. et al. The Stability Enhancement of Nitrile Hydratase from Bordetella petrii by Swapping the C-terminal Domain of β subunit. Appl Biochem Biotechnol 178, 1481–1487 (2016). https://doi.org/10.1007/s12010-015-1961-z

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    • DOI: https://doi.org/10.1007/s12010-015-1961-z

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