Optimization and characterization of antileukemic l-asparaginase produced by Fusarium solani endophyte

• Published in: AMB Express 2023-09, Vol.13 (1), p.96-12, Article 96
• Main Authors: Osama, Sarah, El-Sherei, Moshera M., Al-Mahdy, Dalia A., Bishr, Mokhtar, Salama, Osama, Raafat, Marwa M.
• Format: Article
• Language: English
• Subjects: Acute monocytic leukemia; Ammonia; Anticancer properties; Apoptosis; Asparaginase; Asparagine; Aspartic acid; Biomedical and Life Sciences; Biotechnology; Characterization; Cytotoxicity; Endophytes; Enzymatic activity; Enzyme activity; Enzymes; Fermentation; Fibroblasts; Fusarium; Fusarium solani; Gene banks; Hedera helix; Herbal medicine; L-asparaginase; Leukemia; Life Sciences; Medicinal plants; Microbial Genetics and Genomics; Microbiology; Monocytes; Monocytic leukemia; Optimization; Original; Original Article; Skin tests; Thermal stability; Toxicity; Toxicity testing
• ISSN: 2191-0855; 2191-0855
• DOI: 10.1186/s13568-023-01602-2

l -asparaginase is an antileukemic enzyme that hydrolyzes l -asparagine into l -aspartic acid and ammonia, causing cell starvation and apoptosis in susceptible leukemic cell populations. Currently, l -asparaginase obtained from bacterial sources is constrained by several issues, including lesser productivity, stability, selectivity, and higher toxicity. The goal of this study is to provide fungal l -asparaginase with in-vitro effectiveness towards different human carcinomas. l -asparaginase from endophytic Fusarium solani (Gene Bank accession number MW209717) isolated from the roots of the medicinal plant Hedera helix L. was characterized and optimized experimentally for maximum l -asparaginase production in addition to evaluating its subsequent cytotoxicity towards acute monocytic leukemia and human skin fibroblast cell lines. The enzyme production was maximized using potato dextrose media (15.44 IU/ml/hr) at the 5th and 6th days of fermentation with incubation temperature 30 °C, 3% asparagine, 150–180 rpm agitation rate and a 250 ml flask. Enzyme characterization studies revealed that the enzyme maintained its thermal stability with temperatures up to 60 °C. However, its optimal activity was achieved at 35 °C. On measuring the enzymatic activity at various temperatures and different pH, maximum enzyme activity was recorded at 40 °C and pH 8 using 0.1 M asparagine concentration. Results also revealed promising cytotoxic activity against acute monocytic leukemia with IC 50  = 3.66 µg/ml and low cytotoxicity against tested normal human skin fibroblast cell line which suggested that it might have selective toxicity, and consequently it could be used as a less toxic alternative to the current formulations.