Cloning and Characterization of Chitin Deacetylase from <i>Euphausia superba</i>
Chitin deacetylase (CDA) can catalyze the deacetylation of chitin to produce chitosan. In this study, we identified and characterized a chitin deacetylase gene from <i>Euphausia superba</i> (<i>EsCDA-9k</i>), and a soluble recombinant protein chitin deacetylase from <i>...
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MDPI AG
2024-02-01
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| Series: | International Journal of Molecular Sciences |
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| author | Xutong Wang Jiahao Tan Huaying Zou Fang Wang Jiakun Xu |
| author_facet | Xutong Wang Jiahao Tan Huaying Zou Fang Wang Jiakun Xu |
| author_sort | Xutong Wang |
| collection | DOAJ |
| description | Chitin deacetylase (CDA) can catalyze the deacetylation of chitin to produce chitosan. In this study, we identified and characterized a chitin deacetylase gene from <i>Euphausia superba</i> (<i>EsCDA-9k</i>), and a soluble recombinant protein chitin deacetylase from <i>Euphausia superba</i> of molecular weight 45 kDa was cloned, expressed, and purified. The full-length cDNA sequence of <i>Es</i>CDA-9k was 1068 bp long and encoded 355 amino acid residues that contained the typical domain structure of carbohydrate esterase family 4. The predicted three-dimensional structure of <i>Es</i>CDA-9k showed a 67.32% homology with <i>Penaeus monodon</i>. Recombinant chitin deacetylase had the highest activity at 40 °C and pH 8.0 in Tris-HCl buffer. The enzyme activity was enhanced by metal ions Co<sup>2+</sup>, Fe<sup>3+</sup>, Ca<sup>2+</sup>, and Na<sup>+</sup>, while it was inhibited by Zn<sup>2+</sup>, Ba<sup>2+</sup>, Mg<sup>2+</sup>, and EDTA. Molecular simulation of <i>Es</i>CDA-9k was conducted based on sequence alignment and homology modeling. The <i>Es</i>CDA-9k F18G mutant showed a 1.6-fold higher activity than the wild-type enzyme. In summary, this is the first report of the cloning and heterologous expression of the chitin deacetylase gene in <i>Euphausia superba</i>. The characterization and function study of <i>Es</i>CDA-9k will serve as an important reference point for future application. |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-5f2915989d614134846c226760b82e1f2024-02-23T15:19:39ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672024-02-01254207510.3390/ijms25042075Cloning and Characterization of Chitin Deacetylase from <i>Euphausia superba</i>Xutong Wang0Jiahao Tan1Huaying Zou2Fang Wang3Jiakun Xu4College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, ChinaCollege of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, ChinaCollege of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, ChinaState Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Laboratory for Marine Drugs and Byproducts of Pilot National Lab for Marine Science and Technology, Qingdao 266071, ChinaState Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Laboratory for Marine Drugs and Byproducts of Pilot National Lab for Marine Science and Technology, Qingdao 266071, ChinaChitin deacetylase (CDA) can catalyze the deacetylation of chitin to produce chitosan. In this study, we identified and characterized a chitin deacetylase gene from <i>Euphausia superba</i> (<i>EsCDA-9k</i>), and a soluble recombinant protein chitin deacetylase from <i>Euphausia superba</i> of molecular weight 45 kDa was cloned, expressed, and purified. The full-length cDNA sequence of <i>Es</i>CDA-9k was 1068 bp long and encoded 355 amino acid residues that contained the typical domain structure of carbohydrate esterase family 4. The predicted three-dimensional structure of <i>Es</i>CDA-9k showed a 67.32% homology with <i>Penaeus monodon</i>. Recombinant chitin deacetylase had the highest activity at 40 °C and pH 8.0 in Tris-HCl buffer. The enzyme activity was enhanced by metal ions Co<sup>2+</sup>, Fe<sup>3+</sup>, Ca<sup>2+</sup>, and Na<sup>+</sup>, while it was inhibited by Zn<sup>2+</sup>, Ba<sup>2+</sup>, Mg<sup>2+</sup>, and EDTA. Molecular simulation of <i>Es</i>CDA-9k was conducted based on sequence alignment and homology modeling. The <i>Es</i>CDA-9k F18G mutant showed a 1.6-fold higher activity than the wild-type enzyme. In summary, this is the first report of the cloning and heterologous expression of the chitin deacetylase gene in <i>Euphausia superba</i>. The characterization and function study of <i>Es</i>CDA-9k will serve as an important reference point for future application.https://www.mdpi.com/1422-0067/25/4/2075chitin deacetylase<i>Euphausia superba</i>heterologous expressionenzymatic propertiesmolecular simulation |
| spellingShingle | Xutong Wang Jiahao Tan Huaying Zou Fang Wang Jiakun Xu Cloning and Characterization of Chitin Deacetylase from <i>Euphausia superba</i> International Journal of Molecular Sciences chitin deacetylase <i>Euphausia superba</i> heterologous expression enzymatic properties molecular simulation |
| title | Cloning and Characterization of Chitin Deacetylase from <i>Euphausia superba</i> |
| title_full | Cloning and Characterization of Chitin Deacetylase from <i>Euphausia superba</i> |
| title_fullStr | Cloning and Characterization of Chitin Deacetylase from <i>Euphausia superba</i> |
| title_full_unstemmed | Cloning and Characterization of Chitin Deacetylase from <i>Euphausia superba</i> |
| title_short | Cloning and Characterization of Chitin Deacetylase from <i>Euphausia superba</i> |
| title_sort | cloning and characterization of chitin deacetylase from i euphausia superba i |
| topic | chitin deacetylase <i>Euphausia superba</i> heterologous expression enzymatic properties molecular simulation |
| url | https://www.mdpi.com/1422-0067/25/4/2075 |
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