Characterization of novel cold-active chitin deacetylase for green production of bioactive chitosan
Abstract A Novel cold-active chitin deacetylase from Shewanella psychrophila WP2 (SpsCDA) was overexpressed in Escherichia coli BL21 and employed for deacetylation of chitin to chitosan. The produced chitosan was characterized, and its antifungal activity was investigated against Fusarium oxysporum....
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2025-01-01
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| Online Access: | https://doi.org/10.1186/s13568-024-01804-2 |
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| author | Mohamed N. Abd El-Ghany Salwa A. Hamdi Ahmed K. Zahran Mustafa A. Abou-Taleb Abdallah M. Heikel Muhammed T. Abou El-Kheir Mohamed G. Farahat |
| author_facet | Mohamed N. Abd El-Ghany Salwa A. Hamdi Ahmed K. Zahran Mustafa A. Abou-Taleb Abdallah M. Heikel Muhammed T. Abou El-Kheir Mohamed G. Farahat |
| author_sort | Mohamed N. Abd El-Ghany |
| collection | DOAJ |
| description | Abstract A Novel cold-active chitin deacetylase from Shewanella psychrophila WP2 (SpsCDA) was overexpressed in Escherichia coli BL21 and employed for deacetylation of chitin to chitosan. The produced chitosan was characterized, and its antifungal activity was investigated against Fusarium oxysporum. The purified recombinant SpsCDA appeared as a single band on SDS-PAGE at approximately 60 kDa, and its specific activity was 92 U/mg. The optimum temperature and pH of SpsCDA were 15 °C and 8.0, respectively, and the enzyme activity was significantly enhanced in the presence of NaCl. The bioconversion of chitin to chitosan by SpsCDA was accomplished in 72 h, and the chitosan yield was 69.2%. The solubility of chitosan was estimated to be 73.4%, and the degree of deacetylation was 78.1%. The estimated molecular weight of the produced chitosan was 224.7 ± 8.4 kDa with a crystallinity index (CrI) value of 18.75. Moreover, FTIR and XRD spectra revealed the characteristic peaks for enzymatically produced chitosan compared with standard chitosan, indicating their structural similarity. The produced chitosan inhibited spore germination of F. oxysporum with a minimum inhibitory concentration (MIC) of 1.56 mg/mL. The potential antifungal effect of chitosan is attributed to the inhibition of spore germination accompanied by ultrastructural damage of membranes and leakage of cellular components, as evidenced by transmission electron microscopy (TEM), and accumulation of reactive oxygen species (ROS) that was confirmed by fluorescence microscopy. This study shed light on the cold-active chitin deacetylase from S. psychrophila and provides a candidate enzyme for the green preparation of chitosan. |
| format | Article |
| id | doaj-art-bf2eb4f930aa43ee93485eef5d646d9e |
| institution | Kabale University |
| issn | 2191-0855 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | AMB Express |
| spelling | doaj-art-bf2eb4f930aa43ee93485eef5d646d9e2025-01-05T12:47:30ZengSpringerOpenAMB Express2191-08552025-01-0115111410.1186/s13568-024-01804-2Characterization of novel cold-active chitin deacetylase for green production of bioactive chitosanMohamed N. Abd El-Ghany0Salwa A. Hamdi1Ahmed K. Zahran2Mustafa A. Abou-Taleb3Abdallah M. Heikel4Muhammed T. Abou El-Kheir5Mohamed G. Farahat6Botany and Microbiology Department, Faculty of Science, Cairo UniversityZoology Department, Faculty of Science, Cairo UniversityBiotechnology / Molecular Biochemistry Program, Faculty of Science, Cairo UniversityBiotechnology / Molecular Biochemistry Program, Faculty of Science, Cairo UniversityBiotechnology / Molecular Biochemistry Program, Faculty of Science, Cairo UniversityBiotechnology / Molecular Biochemistry Program, Faculty of Science, Cairo UniversityBotany and Microbiology Department, Faculty of Science, Cairo UniversityAbstract A Novel cold-active chitin deacetylase from Shewanella psychrophila WP2 (SpsCDA) was overexpressed in Escherichia coli BL21 and employed for deacetylation of chitin to chitosan. The produced chitosan was characterized, and its antifungal activity was investigated against Fusarium oxysporum. The purified recombinant SpsCDA appeared as a single band on SDS-PAGE at approximately 60 kDa, and its specific activity was 92 U/mg. The optimum temperature and pH of SpsCDA were 15 °C and 8.0, respectively, and the enzyme activity was significantly enhanced in the presence of NaCl. The bioconversion of chitin to chitosan by SpsCDA was accomplished in 72 h, and the chitosan yield was 69.2%. The solubility of chitosan was estimated to be 73.4%, and the degree of deacetylation was 78.1%. The estimated molecular weight of the produced chitosan was 224.7 ± 8.4 kDa with a crystallinity index (CrI) value of 18.75. Moreover, FTIR and XRD spectra revealed the characteristic peaks for enzymatically produced chitosan compared with standard chitosan, indicating their structural similarity. The produced chitosan inhibited spore germination of F. oxysporum with a minimum inhibitory concentration (MIC) of 1.56 mg/mL. The potential antifungal effect of chitosan is attributed to the inhibition of spore germination accompanied by ultrastructural damage of membranes and leakage of cellular components, as evidenced by transmission electron microscopy (TEM), and accumulation of reactive oxygen species (ROS) that was confirmed by fluorescence microscopy. This study shed light on the cold-active chitin deacetylase from S. psychrophila and provides a candidate enzyme for the green preparation of chitosan.https://doi.org/10.1186/s13568-024-01804-2Chitin extractionChitin deacetylasePsychrophilic bacteriaAntifungal chitosanSpore germinationMembrane permeability |
| spellingShingle | Mohamed N. Abd El-Ghany Salwa A. Hamdi Ahmed K. Zahran Mustafa A. Abou-Taleb Abdallah M. Heikel Muhammed T. Abou El-Kheir Mohamed G. Farahat Characterization of novel cold-active chitin deacetylase for green production of bioactive chitosan AMB Express Chitin extraction Chitin deacetylase Psychrophilic bacteria Antifungal chitosan Spore germination Membrane permeability |
| title | Characterization of novel cold-active chitin deacetylase for green production of bioactive chitosan |
| title_full | Characterization of novel cold-active chitin deacetylase for green production of bioactive chitosan |
| title_fullStr | Characterization of novel cold-active chitin deacetylase for green production of bioactive chitosan |
| title_full_unstemmed | Characterization of novel cold-active chitin deacetylase for green production of bioactive chitosan |
| title_short | Characterization of novel cold-active chitin deacetylase for green production of bioactive chitosan |
| title_sort | characterization of novel cold active chitin deacetylase for green production of bioactive chitosan |
| topic | Chitin extraction Chitin deacetylase Psychrophilic bacteria Antifungal chitosan Spore germination Membrane permeability |
| url | https://doi.org/10.1186/s13568-024-01804-2 |
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