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Structure-based mining of a chitosanase with distinctive degradation mode and product specificity

  • Biotechnologically Relevant Enzymes and Proteins
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Abstract

Chitosan derivates with varying degrees of polymerization (DP) have attracted great concern due to their excellent biological activities. Increasing the abundance of chitosanases with different degradation modes contributes to revealing their catalytic mechanisms and facilitating the production of chitosan derivates. However, the identification of endo-chitosanases capable of producing chitobiose and D-glucosamine (GlcN) from chitosan substrates has remained elusive. Herein, an endo-chitosanase (CsnCA) belonging to the GH46 family was identified based on structural analysis in phylogenetic evolution. Moreover, we demonstrate that CsnCA acts in a random endo-acting manner, producing chitosan derivatives with DP ≤ 2. The in-depth analysis of CsnCA revealed that (GlcN)3 serves as the minimal substrate, undergoing cleavage in the mode that occupies the subsites − 2 to + 1, resulting in the release of GlcN. This study succeeded in discovering a chitosanase with distinctive degradation modes, which could facilitate the mechanistic understanding of chitosanases, further empowering the production of chitosan derivates with specific DP.

Graphical Abstract

Key points

• Structural docking and evolutionary analysis guide to mining the chitosanase.

• The endo-chitosanase exhibits a unique GlcN-producing cleavage pattern.

• The cleavage direction of chitosanase to produce GlcN was identified.

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The data generated or analyzed during this study are included in this published article and its supplementary information files.

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Funding

This work was supported by the National Natural Science Foundation of China (U21A20271 and 31922072) and China Agriculture Research System of MOF and MARA (CARS-48).

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Authors and Affiliations

  1. Qingdao Key Laboratory of Food Biotechnology, College of Food Science and Engineering, Ocean University of China, Qingdao, 266404, China

    Haipeng Su, Jianan Sun, Chaoran Guo, Yongzhen Wang, Hao Dong & Xiangzhao Mao

  2. Key Laboratory of Biological Processing of Aquatic Products, China National Light Industry, Qingdao, 266404, China

    Haipeng Su, Jianan Sun, Chaoran Guo, Yongzhen Wang, Hao Dong & Xiangzhao Mao

  3. Consiglio Nazionale Delle Ricerche, Istituto Di Scienze E Tecnologie Chimiche “Giulio Natta”, Via Bianco Mario, 9, 20131, Milan, Italy

    Francesco Secundo

  4. Laboratory for Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266237, China

    Xiangzhao Mao

Authors
  1. Haipeng Su
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  2. Jianan Sun
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  3. Chaoran Guo
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  4. Yongzhen Wang
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  5. Francesco Secundo
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  6. Hao Dong
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  7. Xiangzhao Mao
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Contributions

H.P.S and H.D conceived and designed the research. H.P.S and Y.Z.W conducted experiments. C.R.G contributed new reagents or analytical tools. H.P.S and J.S performed the literature search and analyzed the data. H.P.S and H.D wrote the manuscript. F.S reviewed and edited the manuscript. X.Z.M administrated the project and provided funds. All authors read and approved the manuscript.

Corresponding author

Correspondence to Hao Dong.

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Su, H., Sun, J., Guo, C. et al. Structure-based mining of a chitosanase with distinctive degradation mode and product specificity. Appl Microbiol Biotechnol 107, 6859–6871 (2023). https://doi.org/10.1007/s00253-023-12741-8

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  • DOI: https://doi.org/10.1007/s00253-023-12741-8

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