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

View Article: PubMed Central - PubMed

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−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.

No MeSH data available.


Effect of different incubation periods on L-asparaginase activity.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC5015098&req=5

f10: Effect of different incubation periods on L-asparaginase activity.

Mentions: The L-asparaginase activity (Fig. 10), increased as the incubation time increased up to 30 min (L-asparaginase activity of 34.128 IU). After which only a slight decrease in L-asparaginase activity was observed. El-Bessoumy et al.22 reported that, maximum activity of L-asparaginase purified from Pseudomonas aeruginosa 50071 was at 30 min. In addition, the effect of incubation time on the activity of purified L-asparaginase from Streptomyces noursei showed that the activity reached its maximum at 35 min15. The decrease in L-asparaginase activity was observed after longer period of incubation with substrate. The decrease may due to the product inhibition.


Purification, characterization, cytotoxicity and anticancer activities of L-asparaginase, anti-colon cancer protein, from the newly isolated alkaliphilic Streptomyces fradiae NEAE-82
Effect of different incubation periods on L-asparaginase activity.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5015098&req=5

f10: Effect of different incubation periods on L-asparaginase activity.
Mentions: The L-asparaginase activity (Fig. 10), increased as the incubation time increased up to 30 min (L-asparaginase activity of 34.128 IU). After which only a slight decrease in L-asparaginase activity was observed. El-Bessoumy et al.22 reported that, maximum activity of L-asparaginase purified from Pseudomonas aeruginosa 50071 was at 30 min. In addition, the effect of incubation time on the activity of purified L-asparaginase from Streptomyces noursei showed that the activity reached its maximum at 35 min15. The decrease in L-asparaginase activity was observed after longer period of incubation with substrate. The decrease may due to the product inhibition.

View Article: PubMed Central - PubMed

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−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.

No MeSH data available.