Purification, characterization, cytotoxicity and anticancer activities of L-asparaginase, anti-colon cancer protein, from the newly isolated alkaliphilic Streptomyces fradiae NEAE-82.

ABSTRACT: 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(-1)min(-1), respectively. The half-life time (T1/2) was 184.91?min at 50?°?, while being 179.53?min at 60?°?. 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.

SUBMITTER: El-Naggar NE 

PROVIDER: S-EPMC5015098 | biostudies-literature | 2016 Sep

REPOSITORIES: biostudies-literature