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Gene cloning, functional expression and secretion of the S-layer protein SgsE from Geobacillus stearothermophilus NRS 2004/3a in Lactococcus lactis
Article first published online: 9 JAN 2006
DOI: 10.1016/j.femsle.2004.10.036
1574-6968/asset/cover.gif?v=1&s=069bb2c224e243333d4486580fbd90d9ebeffa6b "FEMS Microbiology Letters")
Additional Information
How to Cite
Novotny, R., Scheberl, A., Giry-Laterriere, M., Messner, P. and Schäffer, C. (2005), Gene cloning, functional expression and secretion of the S-layer protein SgsE from Geobacillus stearothermophilus NRS 2004/3a in Lactococcus lactis. FEMS Microbiology Letters, 242: 27–35. doi: 10.1016/j.femsle.2004.10.036
Publication History
- Issue published online: 9 JAN 2006
- Article first published online: 9 JAN 2006
- Received 19 August 2004, Revised 30 September 2004, Accepted 19 October 2004
- Abstract
- Article
- References
- Cited By
Keywords:
- S-layer protein;
- Geobacillus stearothermophilus;
- Functional expression;
- Secretion;
- Lactococcus lactis
Abstract
The ∼93-kDa surface layer protein SgsE of Geobacillus stearothermophilus NRS 2004/3a forms a regular crystalline array providing a nanopatterned matrix for the future display of biologically relevant molecules. Lactococcus lactis NZ9000 was established as a safe expression host for the controlled targeted production of SgsE based on the broad host-range plasmid pNZ124Sph, into which the nisA promoter was introduced. SgsE devoid of its signal peptide-encoding sequence was cloned into the new vector and purified from the cytoplasm at a yield of 220 mg l− of expression culture. Secretion constructs were based on the signal peptide of the Lactobacillus brevis SlpA protein or the L. lactis Usp45 protein, allowing isolation of 95 mg of secreted rSgsE l−1. N-terminal sequencing confirmed correct processing of SgsE in L. lactis NZ9000. The ability of rSgsE to self-assemble in suspension and to recrystallize on solid supports was demonstrated by electron and atomic force microscopy.