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High Level Activity of Recombinant Lysostaphin After Computer Simulation and Additive-Based Refolding

  • Shabnam Sadoogh Abbasian
  • Safieh Soufian
  • Ehsanollah Ghaznavi-Rad
  • Hamid AbtahiEmail author
Article
  • 35 Downloads

Abstract

Lysostaphin is a peptidoglycan hydrolase, produced by Staphylococcus simulans, which has illustrated significant bactericidal activities against Staphylococcus aureus species. Currently, recombination is the common approach of lysostaphin production. However, the recombinant produced lysostaphin shows weak antibacterial activities. The reason can be the aggregation of produced lysostaphin which leads to the destruction of natural protein folding. The most common strategy providing the best situation for correct refolding of the recombinant protein is dialysis. In this study, based on the computer simulations different condition of dialysis was applied to achieve the most significant antibacterial activity. In this study, lysostaphin was expressed in Escherichia coli (E. coli) BL21 (DE3) pLysS cells and purified by affinity chromatography. Various dialysis methods were employed to enable the protein to refold to its natural form. The results of final protein antibacterial activity evidenced the high efficiency of computer simulations estimation ability in predicting best dialysis buffer according to the interaction between protein and buffer additive compounds. Finally, it was confirmed that the buffer containing proline 0.15 M and glucose 0.2 M caused the best lysostaphin refolding. Employing computer simulation before initiating the dialysis process would be a novel efficient and economic pathway of protein folding recovery.

Keywords

Computer simulation Lysostaphin Protein refolding Recombinant protein Staphylococcus aureus 

Notes

Acknowledgements

This Study was conducted with financial assistance from Arak University of Medical Sciences, Iran, and the authors are grateful for the university’s invaluable contribution to this study.

Funding

This study was emanated from a proposal (No: 1379) approved and supported by Arak University of Medical Sciences, Iran. This study was approved by the ethics committee of Arak University of Medical Sciences, Arak, Iran, Ethics code: 91-128-6.

Compliance with Ethical Standards

Conflict of interest

The authors declare that no conflict of interest exists.

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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Shabnam Sadoogh Abbasian
    • 1
  • Safieh Soufian
    • 2
  • Ehsanollah Ghaznavi-Rad
    • 1
  • Hamid Abtahi
    • 1
    Email author
  1. 1.Molecular and Medicine Research Center, School of MedicineArak University of Medical SciencesArakIran
  2. 2.Payame Noor UniversityTehranIran

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