RESUMEN
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 anti-proliferative 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 pharmaceu-tical exploitation in the treatment of leukemia.
RESUMEN
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.
RESUMEN
Bioassay guided isolation of active natural compounds was performed to investigate the anti-tumor potential of the crude extract and isolated compounds from inflorescences of Piper claussenianum. LC-DAD-UV, GC-MS and NMR analyzes revealed the presence of phenolic metabolites in the methanol crude extract. Phytochemical procedures lead to the isolation of the major flavonoids, 2’,6’-dihydroxy-4-methoxychalcone, 5,7- dihydroxyflavanone and 5-methoxy-7-hydroxyflavanone that were assayed for inhibition or viability stimulation of the human breast cancer cell line MCF-7. The results suggest the 2’,6’-dihydroxy-4-methoxychalcone as the biologically active compound in the crude methanol extract of inflorescences from P. claussenianum. The crude extract was found as potential natural source of compounds with breast cancer cell inhibition properties. All isolated compounds have not been described from this species yet.