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Purification,characterization and antiproliferative activity of L-asparaginase from Aspergillus oryzae CCT 3940 with no glutaminase activity

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Asian Pacific Journal of Tropical Biomedicine 2016年第9期6卷 785-794页

作者： Fernanda Furlan Goncalves Dias ana lucia tasca gois ruiz Adriana Della Torre Helia Harumi Sato Department of Food Science School of Food EngineeringUniversity of Campinas Division of Pharmacology and Toxicology Multidisciplinary Center for ChemicalBiological and Agricultural ResearchUniversity of Campinas

Objective:To explore the anti-proliferative activity of purified L-asparaginase from Aspergillus oryzae CCT 3940(***).Methods:L-asparaginase was produced by submerged fermentation and purified to electrophoresis homogeneity by ionic exchanged chromatography in a fast protein liquid chromatographic *** purified enzyme was characterized and used for the antiproliferative assay against nine tumor cell lines and one non-tumor cell ***:The free glutaminase L-asparaginase was purified 28.6 fold.L-asparaginase showed high stability under physiological condition,remaining stable in the p H range7.0–8.0 after 1 h incubation at temperature range 30–45℃.The Km and Vmax values of purified L-asparaginase were estimated as 0.66 mmol/L and 313 IU/mL,*** purified enzyme could inhibit the growth of a broad range of human tumor cell lines at the concentrations ***,the enzyme from *** CCT 3940 could inhibit tumor growth of leukemia cell line(K562) with a total growth inhibition value of(3.2 ± 2.5) IU/mL and did not inhibit the non-carcinogenic human cell line growth at the concentrations ***:The sensitivity of the cells lines to purified L-asparaginase from *** CCT 3940 appeared to be concentration dependent affording a more significant decrease in cell growth than that observed for the commercial L-asparaginase from Escherichia *** L-asparaginase from *** CCT 3940 has a high potential for pharmaceutical exploitation in the treatment of leukemia.

关键词： L-asparaginase Aspergillus oryzae Purification Anti-tumoral activity

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