L-Asparaginase is an antileukemic agent that depletes L-asparagine “an important nutrient for cancer cells” through the hydrolysis of L-asparagine into L-aspartic acid and ammonia leading to leukemia cell starvation and apoptosis in susceptible leukemic cell populations. Moreover currently, bacterial L-asparaginase has been limited by problems of lower productivity, stability, selectivity and a number of toxicities along with the resistance towards bacterial L-asparaginase. Then the current work aimed to provide pure L-asparaginase with in-vitro efficacy against various human carcinomas without adverse effects related to current L-asparaginase formulations. Submerged fermentation (SMF) bioprocess was applied and improved to maximize L-asparaginase production from Fusarium equiseti AHMF4 as alternative sources of bacteria. The enzyme production in SMF was maximized to reach 40.78 U mL −1 at the 7th day of fermentation with initial pH 7.0, incubation temperature 30 °C, 1.0% glucose as carbon source, 0.2% asparagine as nitrogen source, 0.1% alanine as amino acid supplement and 0.1% KH 2 PO 4 . The purification of AHMF4 L-asparaginase yielded 2.67-fold purification and 48% recovery with final specific activity of 488.1 U mg −1 of protein. Purified L-asparaginase was characterized as serine protease enzyme with molecular weight of 45.7 kDa beside stability at neutral pH and between 20 and 40 °C. Interestingly, purified L-asparaginase showed promising DPPH radical scavenging activity (IC 50 69.12 μg mL −1 ) and anti-proliferative activity against cervical epitheloid carcinoma (Hela), epidermoid larynx carcinoma (Hep-2), hepatocellular carcinoma (HepG-2), Colorectal carcinoma (HCT-116), and breast adenocarcinoma (MCF-7) with IC 50 equal to 2.0, 5.0, 12.40, 8.26 and 22.8 μg mL −1 , respectively. The enzyme showed higher activity, selectivity and anti-proliferative activity against cancerous cells along with tiny cytotoxicity toward normal cells (WI-38) which indicates that it has selective toxicity and it could be applied as a less toxic alternative to the current formulations.
In this investigation three ancient Egyptian medical plants; Plantago albicans L., Thymelaea hirsuta (L.) Endl. and Urginea maritima (L.) were chosen to explore their biochemical properties, anticancer and antimycotic activities against clinical dermatophytes. Growing of Trichophyton soudanense, Trichophyton erinacei, Microsporum audouinii, Microsporum gypseum, Microsporum gallinae, Microsporum ferrogenium, Microsporum cookie, Microsporum racemosum, Microsporum persicolor and Microsporum canis were totally inhibited by the 100 µg/mL of P. albicans methanolic extract. Whereas the growth of T. soudanense, T. erinacei, Trichophyton rubrum, Trichophyton tonsurance and Trichophyton mentagrophytes and Epidermophyton floccosum were 100% inhibited by the methanolic extract of T. hirsiuta at 100 µg/mL concentration, however its acetone extract was more active against Microsporum species. The highest fungi toxicity against all dermatophytic fungi was detected in both methanol and acetone extract of U. maritima at a concentration ranged between 50 to 100 µg/mL. Methanolic extract of P. albicans inhibited the viability of HCT-116, HepG-2, MCF-7 and HeLa cell lines by (41%, 40%, 80% and 81%); (30%, 20%, 66% and 70%); (19%, 0%, 49% and 57%); (5%, 0%, 35% and 45%) and (0%, 0%, 21% and 40%) at 25, 50, 100, 200 and 300 µg/mL, respectively. Moreover, the cells death after treatment with the same concentrations of T. hirsuta methanol extract were (10%, 20%, 23% and 60%); (16%, 40%, 58% and 80%); (40%, 50%, 80% and 100%); (51%, 65%, 100% and 100%) and (49%, 82%, 100% and 100%) in HCT-116, HepG-2, MCF-7 and HeLa cells, respectively. Viability of HCT-116, MCF-7 and HepG-2 cell lines was totally suppressed with 100 µg/mL whereas the HeLa cell growth was reduced to 10% and totally killed at 100 and 200 µg/mL, respectively of U. maritima methanolic extract. All data approve the significance of Egyptian ethnomedical plants as potent source of diverse bioactive pharmaceutical metabolites.
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