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Screening for New Surface Anchoring Domains for Lactococcus lactis.
Tina Vida Plavec1,2, Borut Štrukelj1,2, Aleš Berlec1,2
1Department of Biotechnology, Jožef Stefan Institute, Ljubljana, Slovenia.
Frontiers in Microbiology
|August 29, 2019
Summary
Researchers explored new ways to display proteins on Lactococcus lactis. The ChW-containing cAM12 domain showed promise as an alternative to existing methods for bacterial surface display.
Area of Science:
- Microbiology and Biotechnology
- Protein Engineering and Surface Display Systems
Background:
- Displaying recombinant proteins on bacterial surfaces offers significant biotechnological potential.
- Lactococcus lactis is a suitable host for surface display and in vivo delivery of bioactive proteins.
- Surface display typically relies on anchoring domains for cell wall attachment.
Purpose of the Study:
- To investigate alternative protein anchoring domains for surface display on Lactococcus lactis.
- To evaluate novel anchoring domains beyond the commonly used LysM-containing cAcmA.
Main Methods:
- Screened 15 lactococcal or phage-derived anchoring domains from Pfam categories: LPXTG, LysM, CW_1, Cpl-7, WxL, SH3, and ChW.
- Assessed surface display of model proteins (B domain, DARPin I07) using these domains.
- Validated the efficacy of the most promising domain (cAM12) for heterologous protein anchoring.
Main Results:
- LPXTG, LysM, CW_1, and SH3 domains facilitated significant, though lower, surface display compared to cAcmA.
- The ChW-containing cAM12 domain demonstrated surface display comparable to the reference cAcmA.
- cAM12 enabled effective non-covalent anchoring of model proteins and a therapeutic protein (evasin-3).
Conclusions:
- Multiple protein anchoring domains can be effectively utilized in Lactococcus lactis.
- The ChW-containing cAM12 domain represents a highly promising alternative to established methods for bacterial surface display.
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