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Cell wall anchoring of the Streptococcus pyogenes M6 protein in various lactic acid bacteria
J C Piard1, I Hautefort, V A Fischetti
1Laboratoire de Génétique Microbienne, Institut de Biotechnologie, INRA, Jouy-en-Josas, France. piard@biotec.jouy.inra.fr
Abstract:
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.
Insights
Streptococcus pyogenes M6 protein
Area of Science:
- Microbiology
- Molecular Biology
- Biotechnology
Background:
- The M6 protein from Streptococcus pyogenes is a well-studied cell envelope-associated protein.
- Its C-terminal sorting signals can anchor heterologous proteins to the cell wall.
Purpose of the Study:
- To clone and express the emm6 gene encoding the M6 protein in various lactic acid bacteria (LAB).
- To evaluate the feasibility of using the M6 protein's C-terminal anchor for surface display in LAB.
Main Methods:
- Cloning and expression of the emm6 gene in Lactococcus lactis, Lactobacillus fermentum, Lactobacillus sake, and Streptococcus salivarius subsp. thermophilus.
- Analysis of M6 protein cell wall anchoring in the engineered LAB strains.
Main Results:
- Successful expression of the emm6 gene and efficient cell wall anchoring of the M6 protein in all tested LAB strains.
- Nearly all detectable M6 protein was cell wall-associated in lactobacilli.
- Demonstrated feasibility of M6 protein C-terminal anchor for surface display in LAB.
Conclusions:
- The M6 protein's C-terminal anchor is a viable tool for surface display in lactic acid bacteria.
- This approach holds potential for developing novel LAB-based functional probiotics or delivery systems.