Cell wall anchoring of the Streptococcus pyogenes M6 protein in various lactic acid bacteria.
The M6 protein from Streptococcus pyogenes is the best-characterized member of a family of cell envelope-associated proteins. Based on the observation that the C-terminal sorting signals of these proteins can drive cell wall anchoring of heterologous unanchored proteins, we have cloned and expressed the emm6 structural gene for the M6 protein in various lactic acid bacteria (LAB). The emm6 gene was successfully expressed from lactococcal promoters in several Lactococcus lactis strains, an animal-colonizing Lactobacillus fermentum strain, Lactobacillus sake, and Streptococcus salivarius subsp. thermophilus. The M6 protein was efficiently anchored to the cell wall in all strains tested. In lactobacilli, essentially all detectable M6 protein was cell wall associated. These results suggest the feasibility of using the C-terminal anchor moiety of M6 for protein surface display in LAB.
Piard
J C
JC
Laboratoire de Génétique Microbienne, Institut de Biotechnologie, INRA, Jouy-en-Josas, France. piard@biotec.jouy.inra.fr
Hautefort
I
I
Fischetti
V A
VA
Ehrlich
S D
SD
Fons
M
M
Gruss
A
A
D000942:Antigens, Bacterial; D000954:Antigens, Surface / Q000502:physiology; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D002352:Carrier Proteins; D002473:Cell Wall / Q000502:physiology; D003001:Cloning, Molecular; D017931:DNA Primers; D004926:Escherichia coli; D005798:Genes, Bacterial; D007778:Lactobacillus / Q000235:genetics / Q000502:physiology; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D021382:Protein Sorting Signals / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D013045:Species Specificity; D013291:Streptococcus / Q000235:genetics / Q000502:physiology; D013297:Streptococcus pyogenes / Q000235:genetics / Q000502:physiology*
D000942:Antigens, Bacterial; D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D017931:DNA Primers; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; C014689:streptococcal M protein