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Immobilization of Bioactive Protein A from Staphylococcus aureus (SpA) on the Surface of Bacillus subtilis Spores

Molecular Biotechnology volume 57, pages 756–766 (2015)Cite this article

Abstract

Protein A from Staphylococcus aureus (SpA) is a 40–60 kDa cell-wall component, composed of five homologous immunoglobulin (Ig)-binding domains folded into a three-helix bundle. Each of these five domains is able to bind Igs from many different mammalian species. Recombinant SpA is widely used as a component of diagnostic kits for the detection and purification of IgGs from serum or other biological fluids. In this study, purified SpA was adsorbed and covalently linked to Bacillus subtilis spores. Spores are extremely stable cell forms and are considered as an attractive platform to display heterologous proteins. A sample containing about 36 μg of SpA was covalently immobilized on the surface of 4 × 1010 spores. Spore-bound SpA retained its IgG-binding activity, even after seven consecutive binding and washing steps, suggesting that it can be recycled and utilized several times. FACS analysis revealed that spores with covalently attached SpA had significantly improved fluorescence intensities when compared to those of spores with adsorbed SpA, suggesting that the covalent approach is more efficient than sole adsorption regarding protein attachment to the spore surface.

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Acknowledgments

The authors would like to thank the National Institute of Genetic Engineering and Biotechnology (NIGEB) of Iran for providing the necessary equipments and facilities. Funding for the project was provided by the Iran National Science Foundation (Project No. 90004656).

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Affiliations

  1. Department of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran

    Samira Ghaedmohammadi, Garshasb Rigi & Gholamreza Ahmadian

  2. Chemistry and Chemical Engineering Research Center of Iran, Tehran, Iran

    Reza Zadmard

  3. Department of Structural and Functional Biology, Federico II University of Naples, Naples, Italy

    Ezio Ricca

Authors
  1. Samira Ghaedmohammadi
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  2. Garshasb Rigi
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  3. Reza Zadmard
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  4. Ezio Ricca
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  5. Gholamreza Ahmadian
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Correspondence to Gholamreza Ahmadian.

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Ghaedmohammadi, S., Rigi, G., Zadmard, R. et al. Immobilization of Bioactive Protein A from Staphylococcus aureus (SpA) on the Surface of Bacillus subtilis Spores. Mol Biotechnol 57, 756–766 (2015). https://doi.org/10.1007/s12033-015-9868-z

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  • DOI: https://doi.org/10.1007/s12033-015-9868-z

Keywords

  • Bacillus subtilis
  • Covalent binding
  • Hydrophobic interaction
  • Protein A
  • Spore
  • Surface display