Recombinant Escherichia coli strain produces a ZZ domain displaying biopolyester granules suitable for immunoglobulin G purification.
The immunoglobulin G (IgG) binding ZZ domain of protein A from Staphylococcus aureus was fused to the N terminus of the polyhydroxyalkanoate (PHA) synthase from Cupriavidus necator. The fusion protein was confirmed by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry and mediated formation of ZZ domain-displaying PHA granules in recombinant Escherichia coli. The IgG binding capacity of isolated granules was assessed using enzyme-linked immunosorbent assay and could be enhanced by the overproduction of the ZZ-PHA synthase. ZZ-PHA granules enabled efficient purification of IgG from human serum.
Brockelbank
Jane A
JA
Institute of Molecular Biosciences, Massey University, Private Bag 11222, Palmerston North, New Zealand.
Peters
Verena
V
Rehm
Bernd H A
BH
D000217:Acyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001709:Biotechnology / Q000379:methods; D020118:Cupriavidus necator / Q000201:enzymology / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification* / Q000378:metabolism; D008863:Microspheres; D011091:Polyesters / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D013205:Staphylococcal Protein A / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013211:Staphylococcus aureus / Q000378:metabolism
D007074:Immunoglobulin G; D011091:Polyesters; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D000217:Acyltransferases; C077225:poly(3-hydroxyalkanoic acid) synthase