The Protein Journal

, Volume 37, Issue 5, pp 461–471 | Cite as

Expression and Functional Characterization of Pseudomonas aeruginosa Recombinant l.Asparaginase

  • Hesham SaeedEmail author
  • Hadeer Soudan
  • Amany El-Sharkawy
  • Aida Farag
  • Amira Embaby
  • Farid Ataya


Recombinant l.asparaginase, L.ASNase, from Pseudomonas aeruginosa was purified using nickel affinity chromatography. The affinity purified L.ASNase exhibited a protein band with a molecular weight of 72.4 kDa on a native polyacrylamide gel and 36.276 kDa using SDS–PAGE. The activity of the purified L.ASNase was enhanced by Mg2+ and inhibited by Zn2+ at a concentration of 5 mM. The specificity of the recombinant L.ASNase towards different substrates was examined, and it was found that the enzyme showed the highest activity towards l.asparagine. Moreover, the enzyme showed lower activity towards other substrates such as L.glutamine, urea and acrylamide. The in vitro hemolysis assay revealed that the purified L.ASNase did not show hemolysis effect on blood erythrocytes. Serum and trypsin half-life of L.ASNase suggested that the recombinant L.ASNase retained 50% of its initial activity after 90 and 60 min incubation period in serum and trypsin separately.


Expression Chemotherapy Cloning 


Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Hesham Saeed
    • 1
    Email author
  • Hadeer Soudan
    • 1
  • Amany El-Sharkawy
    • 2
  • Aida Farag
    • 2
  • Amira Embaby
    • 1
  • Farid Ataya
    • 3
    • 4
  1. 1.Department of Biotechnology, Institute of Graduate Studies and ResearchAlexandria UniversityAlexandriaEgypt
  2. 2.Marine Biotechnology and Natural Products Extract LaboratoryNational Institute of Oceanography and Fisheries (NIOF)AlexandriaEgypt
  3. 3.Biochemistry Department, College of ScienceKing Saud UniversityRiyadhKingdom of Saudi Arabia
  4. 4.National Research CentreGizaEgypt

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