Journal Article

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

RICCA, EZIO, Rigi, Garshasb, Zadmard, Reza, Ghaedmohammadi, Samira, Ahmadian, Gholamreza

2015

Overview

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.

• Publisher: Springer US,Springer Science and Business Media LLC,Springer Nature B.V
• Subject: adsorption / analytical kits / Applied Microbiology and Biotechnology / Bacillus subtilis / Bacillus subtilis - chemistry / Bacillus subtilis; Covalent binding; Hydrophobic interaction; Protein A; Spore; Surface display; Bacillus subtilis; Immobilized Proteins; Spores, Bacterial; Staphylococcal Protein A; Staphylococcus aureus; Biotechnology; Bioengineering; Biochemistry; Applied Microbiology and Biotechnology; Molecular Biology / Bacteria / Bacterial / Biochemistry / Bioengineering / Biological Techniques / Biotechnology / blood serum / Cell Biology / Chemistry / Chemistry and Materials Science / Covalent binding / fluorescence / Human Genetics / Hydrophobic interaction / Immobilized Proteins / Immobilized Proteins - chemistry / immunoglobulin G / mammals / Molecular Biology / Protein A / Protein Science / Proteins / Spore / Spores / Spores, Bacterial / Spores, Bacterial - chemistry / Staphylococcal Protein A / Staphylococcal Protein A - chemistry / Staphylococcus aureus / Staphylococcus aureus - chemistry / Surface display / washing