Purification, characterization and antiproliferative activity of L-asparaginase from Aspergillus oryzae CCT 3940 with no glutaminase activity
10.1016/j.apjtb.2016.07.007
- Author:
Fernanda Furlan Gonçalves DIAS
1
;
Helia Harumi SATO
1
;
Ana Lúcia Tasca Gois RUIZ
2
;
Adriana Della TORRE
2
Author Information
1. Department of Food Science, School of Food Engineering, University of Campinas
2. Division of Pharmacology and Toxicology, Multidisciplinary Center for Chemical, Biological and Agricultural Research, University of Campinas
- Publication Type:Journal Article
- Keywords:
Anti-tumoral activity;
Aspergillus oryzae;
L-asparaginase;
Purification
- From:Asian Pacific Journal of Tropical Biomedicine
2016;6(9):785-794
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the anti-proliferative activity of purified L-asparaginase from Aspergillus oryzae CCT 3940 (A. oryzae). Methods L-asparaginase was produced by submerged fermentation and purified to electrophoresis homogeneity by ionic exchanged chromatography in a fast protein liquid chromatographic system. The purified enzyme was characterized and used for the antiproliferative assay against nine tumor cell lines and one non-tumor cell line. Results The free glutaminase L-asparaginase was purified 28.6 fold. L-asparaginase showed high stability under physiological condition, remaining stable in the pH range 7.0–8.0 after 1 h incubation at temperature range 30–45 °C. The Km and Vmax values of purified L-asparaginase were estimated as 0.66 mmol/L and 313 IU/mL, respectively. The purified enzyme could inhibit the growth of a broad range of human tumor cell lines at the concentrations studied. Also, the enzyme from A. oryzae 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 studied. Conclusions The sensitivity of the cells lines to purified L-asparaginase from A. oryzae 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 coli. The L-asparaginase from A. oryzae CCT 3940 has a high potential for pharmaceutical exploitation in the treatment of leukemia.
