Antitumor activity of urease-free L-asparaginase with low glutaminase coactivity produced by marine-derived Aspergillus flavus
Makaleler > Tam Makale · 2023
Yazarlar: Zeycan Nurçe, Yüksel GEZGİN, Elif Esin HAMEŞ
BIOCATALYSIS AND AGRICULTURAL BIOTECHNOLOGY, SCI Expanded İndekslerine Giren Dergi, Uluslararası, Scopus, 2023, cilt 54, ss 102958-102958
DOI: 10.1016/j.bcab.2023.102958
Özet
L-asparaginase, which has a high therapeutic effect in various cancer diseases, hydrolyzes L-asparagine to aspartate and ammonia and is mainly used in treating Acute Lymphoblastic Leukaemia. This study aims to investigate the L-asparaginase production potential of marine filamentous fungi and the partial purification and characterization of the L-asparaginase. The rapid plate screening method was used to measure the L-asparaginase activity of 94 marine fungi. L-asparaginase producers were then screened for glutaminase and urease activity. The selected extracellular L-asparaginase producer, Aspergillus flavus MF10, showed 2.82 U/mL L-asparaginase activity in small-scale fermentation. L-asparaginase was concentrated by ultrafiltration followed by acetone precipitation. The specific activity of the enzyme, purified 4,8 fold by partial purification, was determined as 259 U/mg. There were no urease and glutaminase activities in the fermentation broth, but low glutaminase activity (1.27 U/mL) was detected in the concentrated fermentation broth. The optimum pH and temperature of the L-asparaginase were 8.0 and 40 °C, respectively. L-asparaginase was stable at pH 7.0–8.0 and can maintain its activity at 70% at 40 °C after 1 h. While the enzyme decreased the viability of SH-SY5Y (neuroblastoma) to 43% and LNCap (prostate cancer) to 61%, it did not show cytotoxic effects in healthy cell line L929 (fibroblast). It was also determined that L-asparaginase showed low cytotoxicity in HaCaT (keratinocyte) cells. In conclusion, it was shown that urease-free L-asparaginase with low glutaminase coactivity, produced from marine-derived A. flavus MF10, has the potential for medical use.