@article{Elshafei_El-Ghonemy_2021, title={EXTRACELLULAR GLUTAMINASE-FREE L-ASPARAGINASE FROM TRICHODERMA VIRIDE F2: PURIFICATION, BIOCHEMICAL CHARACTERIZATION AND EVALUATION OF ITS POTENTIAL IN MITIGATING ACRYLAMIDE FORMATION IN STARCHY FRIED FOOD}, volume={11}, url={https://office2.jmbfs.org/index.php/JMBFS/article/view/4336}, DOI={10.15414/jmbfs.4336}, abstractNote={<p>L-asparaginase is an antitumor agent that suppresses cancer cell growth by eliminating L-asparagine from malignant cells. However, the intrinsic glutaminase activity is responsible for significant life-threatening adverse effects. Therefore, glutaminase-free L-asparaginase is far required to improve the therapeutic efficacy of L-asparaginase treatment. L-asparaginase was also used to combat the development of acrylamide in foods rich in carbohydrates cooked at high temperatures. Therefore, this study explores the purification and characterization of glutaminase-free L-asparaginase from <em>Trichoderma viride</em> F2 using agro-industrial residues as substrate. The enzyme was purified 36-folds with 688.1 U/mg specific activity and a final yield of 38.9% through ethanol precipitation, gel filtration on Sephadex G-100 followed by Sephadex G-200. The purified L-asparaginase is monomeric with a molecular mass of 57 kDa and exhibited optimum activity at pH 7.5 and 37 °C, which is relatively close to the human body’s internal environment. The purified L-asparaginase showed high affinity and catalytic efficiency towards its natural substrate L-asparagine with <em>K<sub>m</sub></em> and <em>V<sub>max</sub></em> of 1.2 mM and 71.3 U/mL, respectively, and did not exhibit any intrinsic glutaminase activity. Among the salts tested, the univalent cations Na<sup>+</sup> and K<sup>+</sup> enhanced the activity by 145.7% and 163.5%, respectively, while the presence of Ag<sup>+ </sup>and Fe<sup>+3</sup> displayed a considerable loss in activity. The enzyme showed a good anti-oxidant activity with IC<sub>50</sub> of 66.1μg/mL and was able to convert L-asparagine exist in potatoes to L-aspartic acid and ammonia, suggesting its use as anti-carcinogenic agent and as potential food industry candidate to mitigate acrylamide content in starchy fried food.</p>}, number={2}, journal={Journal of microbiology, biotechnology and food sciences}, author={Elshafei, Ali and El-Ghonemy, Dina H}, year={2021}, month={Oct.}, pages={e4336} }