Iranian Biomedical Journal

en Biochemical and Biological Evaluation of an L-Asparaginase from Isolated Escherichia coli MF-107 as an Anti-Tumor Enzyme on MCF7 Cell Line Pharmaceutical Biotechnology Pharmaceutical Biotechnology مقاله کامل Full Length/Original Article Background: One of the most widely used anticancer agents is microbial L-ASNase. Herein, we assessed the biochemical and biological properties of an isolated L-ASNase from a Gram-negative bacteria strain, Escherichia coli MF-107. Methods: Using garden asparagus, we obtained several bacterial isolates. These strains were further screened for L-ASNase activity. A promising bacterial isolate was selected for L-ASNase production and subsequent purification. The molecular weight of purified L-ASNase was determined.  The MTT assay was applied to assess the cytotoxic effect of the purified enzyme. Also, for caspase activity determination and the apoptotic effect of purified enzyme on in cells, we conducted a real-time PCR method. Results: The molecular weight of the enzyme was approximately 37 kDa. In the pH range of 7.5 to 8, the enzyme had considerable stability. At 35 °C, the purified L-ASNase optimum activity was recorded. The cytotoxic effect of the enzyme on treated cells was dose-dependent with an IC50 value of 5.7 IU/ml. The Bax gene expression considerably raised by 5.75-fold (p < 0.001) upon L-ASNase treatment. On the other hand, the anti-apoptotic Bcl-2 gene expression showed a 2.63-fold increase compared to the control (p < 0.05). It was detected that the mRNA levels of caspase-3 and p53 were considerably upregulated (5.93 and 1.85-fold, respectively). We did not find any alternation in the caspase-8 activity of the treated cells compared to untreated cells. Conclusion: In this research, the proliferation of the breast cancer cells remarkably inhibited via the cytotoxic effect of isolated L-ASNase from microbial sources. Apoptosis, Breast cancer, Escherichia coli, MCF7 cell line 279 290 http://ibj.pasteur.ac.ir/browse.php?a_code=A-10-1096-1&slc_lang=en&sid=1 Masoumeh Shahnazari msh.sani57@yahoo.com 0000-0002-1211-6778 No Research and Development Laboratory, Javid Biotechnology Institute, Tehran, Iran Razieh Bigdeli 67bigdeli@gmail.com 0000-0001-7828-3344 No Research and Development Laboratory, Javid Biotechnology Institute, Tehran, Iran Aziz Dashbolaghi dashblaghi@yahoo.com 0000-0001-7216-3309 No Research and Development Laboratory, Javid Biotechnology Institute, Tehran, Iran Reza Ahangari Cohan cohan_r@yahoo.com 0000-0002-0490-4184 No Department of Nanobiotechnology, New Technologies Research Group, Pasteur Institute of Iran, Tehran, Iran Alireza Shoari alireza_shoari@yahoo.com 0000-0003-3842-2156 No Research and Development Laboratory, Javid Biotechnology Institute, Tehran, Iran Hossein Hosseini No Research and Development Laboratory, Javid Biotechnology Institute, Tehran, Iran Davoud Nouri Inanlou d_nouriinanlou@yahoo.com 0000-0002-2966-9985 No Molecular and Cellular Biology Laboratory, Department of Quality Control, Pasteur Institute of Iran, Tehran, Iran Vahid Asgary vahid.asgary@yahoo.com Yes Research and Development Laboratory, Javid Biotechnology Institute, Tehran, Iran