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Discovering and Designing a Chimeric Hyperthermophilic Chitinase for Crystalline Chitin Degradation
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    Research Article

    Discovering and Designing a Chimeric Hyperthermophilic Chitinase for Crystalline Chitin Degradation
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    • Bo Sun
      Bo Sun
      Key Laboratory of Plant Immunity, Joint International Research Laboratory of Soil Health, College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing 210095, People’s Republic of China
      More by Bo Sun
    • Xingchu Zhao
      Xingchu Zhao
      College of Science, Nanjing Agricultural University, Nanjing 210095, People’s Republic of China
      More by Xingchu Zhao
    • Birui Xu
      Birui Xu
      College of Science, Nanjing Agricultural University, Nanjing 210095, People’s Republic of China
      More by Birui Xu
    • Erzheng Su
      Erzheng Su
      College of Light Industry and Food Engineering, Nanjing Forestry University, Nanjing 210037, People’s Republic of China
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    • Andrey Kovalevsky
      Andrey Kovalevsky
      Neutron Scattering Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37831, United States
    • Qirong Shen
      Qirong Shen
      Key Laboratory of Plant Immunity, Joint International Research Laboratory of Soil Health, College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing 210095, People’s Republic of China
      More by Qirong Shen
    • Dongyang Liu
      Dongyang Liu
      Key Laboratory of Plant Immunity, Joint International Research Laboratory of Soil Health, College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing 210095, People’s Republic of China
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    • Qun Wan*
      Qun Wan
      Key Laboratory of Plant Immunity, Joint International Research Laboratory of Soil Health, College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing 210095, People’s Republic of China
      *Email: [email protected]
      More by Qun Wan
    Other Access OptionsSupporting Information (1)

    ACS Sustainable Chemistry & Engineering

    Cite this: ACS Sustainable Chem. Eng. 2023, 11, 12, 4690–4698
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    https://doi.org/10.1021/acssuschemeng.2c07050
    Published March 14, 2023
    Copyright © 2023 American Chemical Society

    Abstract

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    Chitin is one of the most abundant renewable biopolymers on earth. However, it is highly crystalline and recalcitrant to degrade. Here, we report a hyperthermophilic chitinase (ActChi) to directly hydrolyze crystalline chitin at its optimal temperature of 80 °C. It contains a malectin domain, a fibronectin type-III (Fn3) domain, and a catalytic domain (CDchi). Both Fn3 and malectin have the function of chitin binding domain (ChBD) to increase the activity. Fn3 also significantly increases thermostability, but malectin decreases it. To enhance both activity and thermostability, we introduced a heterogeneous and hyperthermophilic ChBD at the N-terminus of CDchi to obtain ChBD-CDchi. The activity of this hybrid enzyme is 201 U/μmol for crystalline chitin, which has increased 400% compared with that of ActChi. In addition, ChBD-CDchi can continuously degrade crystalline chitin for more than 4 days at 70 °C to increase the overall hydrolysis rate. The strategy is a good example of green sustainable degradation for crystalline biopolymer in nature.

    Copyright © 2023 American Chemical Society

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    Supporting Information

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    The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acssuschemeng.2c07050.

    • Ramachandran plot analysis of the homologous model of ActChi and the SDS-PAGE analysis of ActChi and its variants (PDF)

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    Cited By

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    This article is cited by 8 publications.

    1. S. C. Prasanna, M. Balakrishnan. Synthesis and characterization of basalt/bamboo fiber and chitin biopolymer toughened epoxy biocomposite for human prosthetic applications. Polymer Bulletin 2025, 6 https://doi.org/10.1007/s00289-025-05639-0
    2. Jianlin Song, Haipeng Su, Jianan Sun, Xiangzhao Mao. Biodegradation of Crystalline Chitin: A Review of Recent Advancement, Challenges, and Future Study Directions. Journal of Ocean University of China 2024, 23 (5) , 1319-1328. https://doi.org/10.1007/s11802-024-5983-4
    3. Yuan Yu, Sunan Chen, Mingyan Yan, Yinping Li, Min Yang, Xiaofang Liu, Junkui Miao, Xixi Wang, Mengshi Xiao, Haijin Mou, Kailiang Leng. Identification, expression, and characterization of a marine-derived chitinase Ce0303 from Chitiniphilus eburneus YS-30 with exo- and endo-hydrolytic properties. International Journal of Biological Macromolecules 2024, 276 , 133980. https://doi.org/10.1016/j.ijbiomac.2024.133980
    4. Yang Xu, Bo OuYang, Linye Deng, Mei Liao, Tianhui Tang, Dongming Lan, Yonghua Wang. Biochemical characterization of a novel hyperthermophilic chitinase from a deep-sea Thermotogae bacterium. Process Biochemistry 2024, 143 , 60-72. https://doi.org/10.1016/j.procbio.2024.04.031
    5. Shuanghao Yang, Ruirui Feng, Bo Sun, Meizi Lu, Xingchu Zhao, Qirong Shen, Qun Wan. Discovering a thermophilic xylanase and a β-xylosidase for synergistic degradation of corncob. Biomass Conversion and Biorefinery 2024, 311 https://doi.org/10.1007/s13399-024-05822-9
    6. Ziteng Wang, Yijin Cai, Mingxuan Li, Xiaoru Wan, Li Mi, Wenge Yang, Yonghong Hu. Boosting one-step degradation of shrimp shell waste to produce chitin oligosaccharides at smart nanoscale enzyme reactor with liquid-solid system. International Journal of Biological Macromolecules 2024, 268 , 131787. https://doi.org/10.1016/j.ijbiomac.2024.131787
    7. Mohammad K. A. Khan, M. Faisal, V. R. Arun Prakash. Damage investigations on natural fiber-epoxy human prosthetic composites toughened using echinoidea spike β-chitin biopolymer. Biomass Conversion and Biorefinery 2024, 14 https://doi.org/10.1007/s13399-024-05421-8
    8. Susu Han, Wenjiao Zhang, Qiaojuan Yan, Zhengqiang Jiang, Shaoqing Yang. High level production of a novel acidic and thermostable chitinase from Paecilomyces thermophila for the extraction of bioactive components from Ganoderma lucidum spores. Process Biochemistry 2024, 136 , 182-190. https://doi.org/10.1016/j.procbio.2023.11.023

    ACS Sustainable Chemistry & Engineering

    Cite this: ACS Sustainable Chem. Eng. 2023, 11, 12, 4690–4698
    Click to copy citationCitation copied!
    https://doi.org/10.1021/acssuschemeng.2c07050
    Published March 14, 2023
    Copyright © 2023 American Chemical Society

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