Volume 8, Issue 1 (6-2026)
Tabari Biomed Stu Res J 2026, 8(1): 47-56 |
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hashemi juybari F, rezaie S. Recombinant Production, Characterization, and Nano-Immobilization of Bacillus subtilis L Asparaginase in pH-Responsive Chitosan Alginate Nanoparticles. Tabari Biomed Stu Res J 2026; 8 (1) :47-56
URL: http://tbsrj.mazums.ac.ir/article-1-3915-en.html
URL: http://tbsrj.mazums.ac.ir/article-1-3915-en.html
Abstract:
L‑Asparaginase (EC 3.5.1.1) is a therapeutic enzyme that is widely used in the treatment of acute lymphoblastic leukemia (ALL) and also serves as a processing aid in the food industry. However, its clinical application is limited by poor stability, a short plasma half‑life, and significant immunogenicity. In this study, the AsnZ gene encoding L‑asparaginase from Bacillus subtilis was cloned into pET26b(+) and expressed in Escherichia coli BL21(DE3). The recombinant His-tagged enzyme was purified by Ni‑NTA affinity chromatography and characterized biochemically. The purified enzyme exhibited a total activity of 4.63 U/mL and a specific activity of 18.52 U/mg protein at 37 °C and pH 6.0, with optimal activity observed at 37 °C and pH 6.0. The enzyme retained over 80% of its activity between 30‑45 °C and at pH 5.5‑7.0. Subsequently, the recombinant enzyme was successfully immobilized in chitosan‑alginate nanoparticles prepared by ionic gelation, yielding nanoparticles with an average size of 185.4 nm and a zeta potential of +32.7 mV. The nanoparticles exhibited pH-responsive swelling behavior at physiological pH (7.4), reaching approximately 180% within 8 hours, indicating their potential for controlled release. Our results demonstrate that recombinant L‑asparaginase from B. subtilis can be effectively produced in E. coli and immobilized in biodegradable polymeric nanoparticles, offering a promising platform for developing improved therapeutic formulations for leukemia treatment.
Keywords: L Asparaginase, Recombinant production, Bacillus subtilis, Chitosan alginate nanoparticles, Enzyme immobilization, Acute lymphoblastic leukemia
Type of Study: Research (Original) |
Subject:
Biology
Published: 2026/06/20 | ePublished: 2026/06/20
Published: 2026/06/20 | ePublished: 2026/06/20
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