Alcalase immobilization onto chitosan/glutaraldehyde/tripolyphosphate beads obtained by inverse emulsion technique
Enzymes immobilization can efficiently solve limitations of their large-scale application, such as stability and reusability. In this study, Alcalase® 2.4L (protease from Bacillus licheniformis) was covalently immobilized onto chitosan beads obtained by inverse emulsion technique using 1.5% (m/v) of...
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Association of the Chemical Engineers of Serbia
2025-01-01
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| Series: | Chemical Industry and Chemical Engineering Quarterly |
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| Online Access: | https://doiserbia.nb.rs/img/doi/1451-9372/2025/1451-93722400037Z.pdf |
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| author | Žuža-Praštalo Milena Milašinović Nikola Jonović Marko Kalagasidis-Krušić Melina Knežević-Jugović Zorica |
| author_facet | Žuža-Praštalo Milena Milašinović Nikola Jonović Marko Kalagasidis-Krušić Melina Knežević-Jugović Zorica |
| author_sort | Žuža-Praštalo Milena |
| collection | DOAJ |
| description | Enzymes immobilization can efficiently solve limitations of their large-scale application, such as stability and reusability. In this study, Alcalase® 2.4L (protease from Bacillus licheniformis) was covalently immobilized onto chitosan beads obtained by inverse emulsion technique using 1.5% (m/v) of chitosan and 0.67% (v/v) or 1.0% (v/v) of glutaraldehyde (CTPP (1.5/0.67) and CTPP (1.5/1.0)). Afterward, the beads were additionally crosslinked by immersion into 10 % (m/v) tripolyphosphate solution. The parameters studied were enzyme loading, enzyme coupling yield, bead diameter, SEM, biocatalyst activity, and FTIR. The beads had adequate enzyme loading and enzyme coupling yield (Pgmax was 117.1 mg/g dry CTPP 1.5/0.67 and 90.1 mg/g dry CTPP 1.5/1.0, and μmax was 96.7% for both carriers). CTPP (1.5/1.00) beads were smaller (diameter 635.2 ±25.2 mm wet/ 230.4±12.5 mm dry beads) and showed a higher specific activity of 20.1 ± 0.23 IU/mgprotein. The immobilized Alcalase® 2.4L was tested for hydrolyzing egg white and soy proteins. Alcalase® 2.4L, covalently attached to CTTP (1.5/1.0) chitosan beads, is a promising choice for industrial processes involving egg white protein hydrolysis, as the enzyme achieved a notable hydrolysis rate of 26.34 ± 0.879% after 195 minutes. Additionally, it remained effective through five successive applications under practical conditions (50°C, pH 8). |
| format | Article |
| id | doaj-art-18837311c0b14ae8a34c98f804fcd28f |
| institution | OA Journals |
| issn | 1451-9372 2217-7434 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Association of the Chemical Engineers of Serbia |
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| series | Chemical Industry and Chemical Engineering Quarterly |
| spelling | doaj-art-18837311c0b14ae8a34c98f804fcd28f2025-08-20T01:51:57ZengAssociation of the Chemical Engineers of SerbiaChemical Industry and Chemical Engineering Quarterly1451-93722217-74342025-01-0131429530410.2298/CICEQ240401037Z1451-93722400037ZAlcalase immobilization onto chitosan/glutaraldehyde/tripolyphosphate beads obtained by inverse emulsion techniqueŽuža-Praštalo Milena0https://orcid.org/0000-0001-5275-753XMilašinović Nikola1https://orcid.org/0000-0002-2744-4002Jonović Marko2https://orcid.org/0000-0002-9494-6766Kalagasidis-Krušić Melina3https://orcid.org/0000-0003-0517-1952Knežević-Jugović Zorica4https://orcid.org/0000-0003-3009-1698Department of Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, Belgrade, SerbiaDepartment of Forensic Sciences, Faculty of Forensic Sciences and Engineering, University of Criminal Investigation and Police Studies, Belgrade, SerbiaInstitute of Chemistry, Technology and Metallurgy, University of Belgrade, Belgrade, SerbiaDepartment of Organic Chemical Technology, Faculty of Technology and Metallurgy, University of Belgrade, Belgrade, SerbiaDepartment of Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, Belgrade, SerbiaEnzymes immobilization can efficiently solve limitations of their large-scale application, such as stability and reusability. In this study, Alcalase® 2.4L (protease from Bacillus licheniformis) was covalently immobilized onto chitosan beads obtained by inverse emulsion technique using 1.5% (m/v) of chitosan and 0.67% (v/v) or 1.0% (v/v) of glutaraldehyde (CTPP (1.5/0.67) and CTPP (1.5/1.0)). Afterward, the beads were additionally crosslinked by immersion into 10 % (m/v) tripolyphosphate solution. The parameters studied were enzyme loading, enzyme coupling yield, bead diameter, SEM, biocatalyst activity, and FTIR. The beads had adequate enzyme loading and enzyme coupling yield (Pgmax was 117.1 mg/g dry CTPP 1.5/0.67 and 90.1 mg/g dry CTPP 1.5/1.0, and μmax was 96.7% for both carriers). CTPP (1.5/1.00) beads were smaller (diameter 635.2 ±25.2 mm wet/ 230.4±12.5 mm dry beads) and showed a higher specific activity of 20.1 ± 0.23 IU/mgprotein. The immobilized Alcalase® 2.4L was tested for hydrolyzing egg white and soy proteins. Alcalase® 2.4L, covalently attached to CTTP (1.5/1.0) chitosan beads, is a promising choice for industrial processes involving egg white protein hydrolysis, as the enzyme achieved a notable hydrolysis rate of 26.34 ± 0.879% after 195 minutes. Additionally, it remained effective through five successive applications under practical conditions (50°C, pH 8).https://doiserbia.nb.rs/img/doi/1451-9372/2025/1451-93722400037Z.pdfalcalase® 2.4lcovalent immobilizationinverse emulsion techniquechitosan beadstripolyphosphate |
| spellingShingle | Žuža-Praštalo Milena Milašinović Nikola Jonović Marko Kalagasidis-Krušić Melina Knežević-Jugović Zorica Alcalase immobilization onto chitosan/glutaraldehyde/tripolyphosphate beads obtained by inverse emulsion technique Chemical Industry and Chemical Engineering Quarterly alcalase® 2.4l covalent immobilization inverse emulsion technique chitosan beads tripolyphosphate |
| title | Alcalase immobilization onto chitosan/glutaraldehyde/tripolyphosphate beads obtained by inverse emulsion technique |
| title_full | Alcalase immobilization onto chitosan/glutaraldehyde/tripolyphosphate beads obtained by inverse emulsion technique |
| title_fullStr | Alcalase immobilization onto chitosan/glutaraldehyde/tripolyphosphate beads obtained by inverse emulsion technique |
| title_full_unstemmed | Alcalase immobilization onto chitosan/glutaraldehyde/tripolyphosphate beads obtained by inverse emulsion technique |
| title_short | Alcalase immobilization onto chitosan/glutaraldehyde/tripolyphosphate beads obtained by inverse emulsion technique |
| title_sort | alcalase immobilization onto chitosan glutaraldehyde tripolyphosphate beads obtained by inverse emulsion technique |
| topic | alcalase® 2.4l covalent immobilization inverse emulsion technique chitosan beads tripolyphosphate |
| url | https://doiserbia.nb.rs/img/doi/1451-9372/2025/1451-93722400037Z.pdf |
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