Experimental Evaluation of In Silico Selected Signal Peptides for Secretory Expression of Erwinia Asparaginase in Escherichia coli

  • Maryam Yari
  • Mohammad Bagher Ghoshoon
  • Navid Nezafat
  • Younes GhasemiEmail author


Erwinia chrysanthemi asparaginase is an important drug used in cancer treatment, especially in acute lymphoblastic leukemia (ALL). Escherichia coli periplasmic space is an ideal compartment for recombinant expression of certain proteins. To efficient secretion, an appropriate signal sequence should be chosen for each protein individually. In this study, the proper signal sequences for secretory production of Erwinia asparaginase were predicted by in silico methods. Consequently, two signal peptides, OmpA and DsbA, were selected for secretory expression of the enzyme in E. coli. Asparaginase was translocated through the cytoplasmic membrane with either DsbA signal peptide or OmpA signal peptide; Using DsbA signal peptide, 5.95 units of the enzyme was obtained per milliliter of culture media, whereas OmpA signal sequence led to some amount of periplasmic expression. Our study showed that the co-translational signal peptide, DsbA, targeted the asparaginase to cell membrane more efficiently in comparison to the post-translational signal peptide, OmpA. The combination of in silico and experimental approaches provides a way to test a wide variety of signal sequences for secretory production of the enzyme in a time- and cost-effective manner. It is a fundamental step for further studies in the enzyme production process.


Erwinia asparaginase In silico Periplasmic space Secretion Signal peptide 



The authors wish to thank Shiraz University of Medical Sciences for supporting the conduct of this research.

Compliance with Ethical Standards

Conflict of interest

All the authors declare that they have none conflict of interest.

Research Involving Human and Animal Participants

This article does not contain any studies with human participants or animals performed by any of the authors.


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of Medical Biotechnology, School of Advanced Medical Sciences and TechnologiesShiraz University of Medical SciencesShirazIran
  2. 2.Biotechnology Research CenterShiraz University of Medical SciencesShirazIran
  3. 3.Pharmaceutical Science Research CenterShiraz University of Medical SciencesShirazIran
  4. 4.Department of Pharmaceutical Biotechnology, School of PharmacyShiraz University of Medical SciencesShirazIran

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