Functional display of a heterologous protein on the surface of Lactococcus lactis by means of the cell wall anchor of Staphylococcus aureus protein A.
In this study, we showed that the cell wall anchor of protein A from Staphylococcus aureus is functional in the food-grade organism Lactococcus lactis. A fusion protein composed of the lactococcal Usp45 secretion signal peptide, streptavidin monomer, and the S. aureus protein A anchor became covalently attached to the peptidoglycan when expressed in L. lactis. The streptavidin moiety of the fusion protein was functionally exposed at the cellular surface. L. lactis cells expressing the anchored fusion polypeptide could be specifically immobilized on a biotinylated alkaline phosphatase-coated polystyrene support.
Steidler
L
L
Flanders Interuniversity Institute for Biotechnology, Department of Molecular Biology, Universiteit Gent, Belgium.
Viaene
J
J
Fiers
W
W
Remaut
E
E
D000469:Alkaline Phosphatase / Q000378:metabolism; D019921:Biotinylation; D002473:Cell Wall / Q000378:metabolism*; D003001:Cloning, Molecular; D005822:Genetic Vectors; D013294:Lactococcus lactis / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D021382:Protein Sorting Signals / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D013205:Staphylococcal Protein A / Q000235:genetics / Q000378:metabolism*; D019809:Streptavidin / Q000235:genetics / Q000378:metabolism
D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D019809:Streptavidin; D000469:Alkaline Phosphatase