Immobilization and cytotoxic activity evaluation of L-lysine-α-oxidase (LO) enzyme purified from Pseudomonas aeruginosa | ||
| Microbes and Infectious Diseases | ||
| Articles in Press, Accepted Manuscript, Available Online from 14 September 2025 | ||
| Document Type: Original Article | ||
| DOI: 10.21608/mid.2025.393568.2891 | ||
| Authors | ||
| mohammed ahmed abd* ; Asmaa A. Hussein | ||
| Department of Molecular and Medical Biotechnology, College of Biotechnology, Al-Nahrain University, Jadriya, Baghdad 64074, Iraq | ||
| Abstract | ||
| Background: L-lysine-α-oxidase (LO) is an antioxidant flavoenzyme from the L-amino acid oxidase family that plays a significant role in lysine catabolism and exhibits potential anticancer properties. Aim: This study aims to partially purify and immobilize L-lysine α-oxidase from clinical Pseudomonas aeruginosa and to evaluate its cytotoxic effect against cancer and normal cells. Methods: LO was partial purified from Pseudomonas aeruginosa and immobilized using three supports (gelatin, gelatin-sodium alginate, and sodium alginate). Cytotoxic activity of both immobilized and free LO was evaluated on Caco-2 (human colorectal cancer) and HdFn (normal fibroblast) cell lines using the MTT assay. This in vitro experimental study was conducted in accordance with institutional ethical standards for the use of human cell lines. Results: The highest immobilization efficiency (96.2%) was obtained using gelatin-sodium alginate. Free LO reduced Caco-2 cell viability to 39.5% at 400 µg/ml, while immobilized LO reduced it to 51.2% at the same concentration. In HdFn cells, viability decreased to 71.4% (free LO) and 75% (immobilized LO). The IC₅₀ value for free LO against Caco-2 cells was 153.1 µg/ml, compared to 92.35 µg/ml for the immobilized form, indicating higher cytotoxic potency of the immobilized enzyme. Conclusion: L-lysine α-oxidase partial purified from clinical Pseudomonas aeruginosa exhibited selective cytotoxicity against Caco-2 colorectal cancer cells. The immobilization of the enzyme enhanced both its potency and stability, underscoring its potential as a promising candidate for colorectal cancer therapy. | ||
| Keywords | ||
| L-lysine-α-oxidase; Pseudomonas aeruginosa; Immobilization; Cytotoxicity; Cancer therapy | ||
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