ORIGINAL RESEARCH article
Front. Bioeng. Biotechnol.
Sec. Industrial Biotechnology
Volume 13 - 2025 | doi: 10.3389/fbioe.2025.1706906
Immobilized Staphylococcus aureus phospholipase C on calcium alginate-chitosan: improved performance and industrial potential in soybean oil refining
Provisionally accepted
- 1Biochemistry Department, College of Science, King Saud University, Riyadh, Saudi Arabia
- 2Laboratory of Biochemistry and Lipase Enzyme Engineering, ENIS, University of Sfax, Sfax, Tunisia
- 3Cegep de Riviere-du-Loup, Rivière-du-Loup, Canada
- 4King Saud University, Riyadh, Saudi Arabia
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Phospholipases are versatile biocatalysts with wide applications in food and oil industries due to their ability to hydrolyze phospholipids and improve product quality. In this study, production of an extracellular phospholipase C from Staphylococcus aureus (PLCS.a) was optimized, purified, and its immobilization on calcium alginate–chitosan (CAC) was evaluated for soybean oil degumming. Statistical optimization significantly enhanced PLCS.a production showing maximum yields during the exponential growth phase (232.5 U/mL at 34 h) and optimal activity at 37 °C and pH 7.5. Medium composition remarkably influenced enzyme activity, with glucose (2%), BSA (1.5%), Tween 20/80 (0.5-1%), and Zn²⁺/Ca²⁺ (0.1-0.3%) were identified as the most effective enhancers. Immobilization markedly improved enzyme stability, thermal and pH tolerance, and catalytic efficiency compared to the free form. PLCS.a exhibited broad substrate specificity, showing maximum activity toward phosphatidylcholine, and maintained activity in the presence of bile salts and divalent cations. Biotechnological application demonstrated efficient reduction of phosphorus levels in soybean oil from 198 mg/kg to below 2.5 mg/kg within 10 h. Reusability assessment showed that immobilized CAC-PLCS.a retained 100% activity for two cycles, 85.5% after five cycles and 60% after nine cycles, confirming its operational stability. These results highlighted the potential of immobilized PLCS.a as a cost-effective, eco-friendly, and reusable biocatalyst for large-scale vegetable oil refining.
Keywords: phospholipase C, Staphylococcus aureus, Enzyme Immobilization, Calcium alginate–chitosan, Enzyme Stability, Soybean oil degumming, reusability
Received: 16 Sep 2025; Accepted: 13 Oct 2025.
Copyright: © 2025 Alzahrani, KRAYEM, Alonazi, Alshehri, Horchani and Ben Bacha. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Abir Ben Bacha, aalghanouchi@ksu.edu.sa
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