Covalent Immobilization of Dothideomycetes sp. NRC-SSW Chitosanase and Its Application in Chitosan Hydrolysis
Ismail, S., Ismail, S., Hassan, M., Hosny, A., Hashem, A. (2022). Covalent Immobilization of Dothideomycetes sp. NRC-SSW Chitosanase and Its Application in Chitosan Hydrolysis. EKB Journal Management System, 65(1), 397-406. doi: 10.21608/ejchem.2021.80642.4000
Shaymaa Ismail; Siham Ismail; Mohamed El Sayed Hassan; Alaa El-Dein Hosny; Amal Mohamed Hashem. "Covalent Immobilization of Dothideomycetes sp. NRC-SSW Chitosanase and Its Application in Chitosan Hydrolysis". EKB Journal Management System, 65, 1, 2022, 397-406. doi: 10.21608/ejchem.2021.80642.4000
Ismail, S., Ismail, S., Hassan, M., Hosny, A., Hashem, A. (2022). 'Covalent Immobilization of Dothideomycetes sp. NRC-SSW Chitosanase and Its Application in Chitosan Hydrolysis', EKB Journal Management System, 65(1), pp. 397-406. doi: 10.21608/ejchem.2021.80642.4000
Ismail, S., Ismail, S., Hassan, M., Hosny, A., Hashem, A. Covalent Immobilization of Dothideomycetes sp. NRC-SSW Chitosanase and Its Application in Chitosan Hydrolysis. EKB Journal Management System, 2022; 65(1): 397-406. doi: 10.21608/ejchem.2021.80642.4000

Covalent Immobilization of Dothideomycetes sp. NRC-SSW Chitosanase and Its Application in Chitosan Hydrolysis

Egyptian Journal of Chemistry
Article 42, Volume 65, Issue 1, January 2022, Page 397-406  XML PDF (665.51 K)
Document Type: Original Article
DOI: 10.21608/ejchem.2021.80642.4000
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Authors
Shaymaa Ismailorcid 1; Siham Ismail2; Mohamed El Sayed Hassanorcid 3; Alaa El-Dein Hosny4; Amal Mohamed Hashem email orcid 2
1Department of Chemistry of Natural and Microbial Products, Division of Pharmaceutical and Drug Industries, National Research Centre, Cairo, Egypt.
2Chemistry of Natural and Microbial Products Department, National Research Centre, Dokki, Giza, Egypt.
3Center of Excellence, Encapsulation & Nano biotechnology Group, Chemistry of Natural and Microbial Products Department, National Research Center, El Behouth Street, Cairo 12622, Egypt.
4microbiology department, faculty of pharmacy, Cairo university, Cairo, Egypt.
Abstract
Chitosanases are chitosan specific glycosyl hydrolases that can exploit in several biotechnological applications. In the current study, a modified bead of carrageenan and carboxymethyl cellulose was used for the covalent immobilization of Dothideomycetes sp. NRC-SSW chitosanase. The immobilization process was optimized and the immobilization recovery was increased up to 69%. The stepwise different bead formulations were examined under Scanning Electron Microscope, Fourier Transform Infrared Spectroscopy and Thermal Gravimetric analysis, confirming the success of the immobilization process. Additionally different properties including enzyme storage, thermal and pH stabilities were examined and an improvement in all of these properties was estimated. Although the same Vmax value (23.8U/min/mg protein) was recorded for the free and the immobilized enzymes, the Km was 10mg chitosan/ml and 16.67mg chitosan/ml for the immobilized and free form respectively. The activation energy for chitosan hydrolysis using the immobilized enzyme (12.497kJmol−1) was less than the third of that recorded for the free one (42.196kJmol−1). Furthermore, the immobilized enzyme was stable until the 5th cycle at which it retained more than 50% of its original activity. All the tested parameters confirmed that the immobilized Dothideomycetes sp. NRC-SSW chitosanase was more applicable for chitosan hydrolysis than the native enzyme.
Keywords
Chitosanase; Dothideomycetes sp. NRC-SSW; immobilization; chitosan hydrolysis
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