Binding Stability and Catalytic Mechanism of a Novel Chitin Deacetylase from <i>Cytobacillus firmus</i> and Preparation of Chitosan
This study reports the heterologous expression, purification, and functional characterization of a novel chitin deacetylase (CDA) from <i>Cytobacillus firmus</i>. The recombinant <i>Cf</i>CDA exhibited optimal activity at pH 7 and 35 °C, with Mg<sup>2+</sup> enhancing its function. Deep eutectic solvent (DES) pretreatment significantly improved chitin accessibility, enabling <i>Cf</i>CDA to produce chitosan with a high degree of deacetylation (71.2%) and recovery rate (14.2%). Structural analyses confirmed reduced crystallinity and successful acetyl removal. Molecular docking and dynamics simulations identified Asp318, His470, and Trp433 as key catalytic residues with stable binding to (GlcNAc)<sub>4</sub>. These findings highlight <i>Cf</i>CDA's potential for eco-friendly chitosan production and provide a basis for future enzyme engineering.
Authors
- Haoyue Yang (ORCID: https://orcid.org/0000-0001-8867-6504)
- Guantian Li (ORCID: https://orcid.org/0000-0002-3294-2153)
- Kecheng Li (ORCID: https://orcid.org/0000-0001-8862-9943)
- Song Liu (ORCID: https://orcid.org/0000-0002-3941-4337)
- Bicheng Liang
- Zongji Wang (ORCID: https://orcid.org/0000-0002-5107-6246)
- Huahua Yu (ORCID: https://orcid.org/0000-0003-3213-6212)
- Ronge Xing (ORCID: https://orcid.org/0000-0001-8376-023X)
Institutions
- Linyi University (CN)
- Chinese Academy of Sciences (CN)
- Ocean Institute (US)
- Institute of Oceanology (CN)
- Qingdao National Laboratory for Marine Science and Technology (CN)
- Regenerative Medicine Institute (MX)
- Institute of Oceanology (BG)
- University of Chinese Academy of Sciences (CN)
Publication Details
- Journal
- Journal of Agricultural and Food Chemistry
- Published
- 2025-09-22
- DOI
- https://doi.org/10.1021/acs.jafc.5c08532
- Citations
- 1
- Primary Topic
- Studies on Chitinases and Chitosanases
- Type
- article
- Field-Weighted Citation Impact
- 0.81