Purification, characterization, cytotoxicity and anticancer activities of L-asparaginase, anti-colon cancer protein, from the newly isolated alkaliphilic Streptomyces fradiae NEAE-82
| Autor: | Sahar F. Deraz, Sara M. El-Ewasy, Hoda M. Soliman, Nehal M. El-Deeb, Noura El-Ahmady El-Naggar |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0106 biological sciences
0301 basic medicine Ion chromatography Antineoplastic Agents 01 natural sciences Streptomyces Article Substrate Specificity 03 medical and health sciences chemistry.chemical_compound Enzyme activator Anti-Infective Agents 010608 biotechnology Enzyme Stability Asparaginase Ammonium sulfate precipitation Enzyme Assays chemistry.chemical_classification Multidisciplinary biology Streptomyces fradiae biology.organism_classification Enzyme assay Actinobacteria Enzyme Activation 030104 developmental biology Enzyme DEAE-Sepharose chemistry Biochemistry RNA Ribosomal Colonic Neoplasms biology.protein Databases Nucleic Acid |
| Zdroj: | Scientific Reports |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/srep32926 |
| Popis: | L-asparaginase is an important enzyme as therapeutic agents used in combination with other drugs in the treatment of acute lymphoblastic leukemia. A newly isolated actinomycetes strain, Streptomyces sp. NEAE-82, was potentially producing extracellular L-asparaginase, it was identified as Streptomyces fradiae NEAE-82, sequencing product was deposited in the GenBank database under accession number KJ467538. L-asparaginase was purified from the crude enzyme using ammonium sulfate precipitation, dialysis and ion exchange chromatography using DEAE Sepharose CL-6B. Further the kinetic studies of purified enzyme were carried out. The optimum pH, temperature and incubation time for maximum L-asparaginase activity were found to be 8.5, 40 °C and 30 min, respectively. The optimum substrate concentration was found to be 0.06 M. The Km and Vmax of the enzyme were 0.01007 M and 95.08 Uml−1min−1, respectively. The half-life time (T1/2) was 184.91 min at 50 °С, while being 179.53 min at 60 °С. The molecular weight of the subunits of L-asparaginase was found to be approximately 53 kDa by SDS–PAGE analysis. The purified L-asparaginase showed a final specific activity of 30.636 U/mg protein and was purified 3.338-fold. The present work for the first time reported more information in the production, purification and characterization of L-asparaginase produced by newly isolated actinomycetes Streptomyces fradiae NEAE-82. |
| Databáze: | OpenAIRE |
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