ALCALASE IMMOBILIZATION ONTO CHITOSAN/GLUTARALDEHYDE/TRIPOLYPHOSPHATE BEADS OBTAINED BY INVERSE EMULSION TECHNIQUE

Original scientific paper

Authors

  • Milena Žuža Praštalo Department of Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11000 Belgrade, Serbia
  • Nikola Milašinović Department of Forensic Sciences, Faculty of Forensic Sciences and Engineering, University of Criminal Investigation and Police Studies, Cara Dušana 196, 11080 Belgrade, Serbia https://orcid.org/0000-0002-2744-4002
  • Marko Jonović Institute of Chemistry, Technology and Metallurgy, University of Belgrade, Njegoševa 12, 11000 Belgrade, Serbia https://orcid.org/0000-0002-9494-6766
  • Melina Kalagasidis-Krušić Department of Organic Chemical Technology, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11000 Belgrade, Serbia
  • Zorica Knežević-Jugović Department of Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11000 Belgrade, Serbia

DOI:

https://doi.org/10.2298/CICEQ240401037Z

Keywords:

Alcalase® 2.4L, covalent immobilization, inverse emulsion technique, chitosan beads, tripolyphosphate

Abstract

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).

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Published

10.12.2024 — Updated on 16.04.2025

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Vol. 31 No. 4 (2025)

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Copyright (c) 2023 Milena Žuža Praštalo , Nikola Milašinović, Marko Jonović, Melina Kalagasidis-Krušić , Zorica Knežević-Jugović

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How to Cite

ALCALASE IMMOBILIZATION ONTO CHITOSAN/GLUTARALDEHYDE/TRIPOLYPHOSPHATE BEADS OBTAINED BY INVERSE EMULSION TECHNIQUE: Original scientific paper. (2025). Chemical Industry & Chemical Engineering Quarterly, 31(4), 295-304. https://doi.org/10.2298/CICEQ240401037Z

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