Surface display of respiratory syncytial virus glycoproteins in Lactococcus lactis NZ9000

Abstract

Aims: A system for displaying heterologous respiratory syncytial virus (RSV) glycoproteins on the surface of Lactococcus lactis NZ9000 was developed. Methods and Results: Fusion of the USP45 signal peptide and the cA (C terminus of the peptidoglycan‐binding) domains of AcmA, a major autolysin from L. lactis, to the N‐ and C‐terminal of the target proteins, respectively, was carried out. The target protein was the major immunogenic domain of either the F (40·17‐kDa) or G (11·49‐kDa) glycoprotein domains of the RSV. Whole‐cell ELISA readings obtained after 24 h of induction showed an increase in protein expression as the cA domain repeats increased, for the G glycoprotein of RSV. On the other hand, the F glycoprotein indicated decreasing expression levels as the number of cA domain repeats increased. The difference in the expression levels of the F and G domains may be attributed to the different sizes of the antigenic domains. Conclusions: The size and properties of the target proteins are vital in determining the amount of antigenic domains being displayed on the surface of live cells. Significance and Impact of the Study: The system demonstrated here can aid in the utilization of the generally regarded as safe (GRAS) bacteria L. lactis, as a vaccine delivery vehicle to surface display the antigenic proteins of RSV.

Genetically Engineered Lactococcus lactis Protect against House Dust Mite Allergy in a BALB/c Mouse Model

“…The resulting plasmid verified by DNA sequencing was designated pNZ8148-ID, and the obtained L. lactis strain was named LL-I. To construct the other two plasmids, pNZ8148-ED and pNZ8148-WD, the signal peptide of Usp 45 and the anchor gene fragment of N-acetylmuramidase (acmA) on the L. lactis MG1363 genome were amplified using the respective primers [15] . Both plasmids were manipulated with a similar protocol in the correct order to obtain recombinant L. lactis strains LL-E and LL-W, respectively.…”

Surface display system for probiotics and its application in aquaculture

“…2008), while this was not the case for all kinds of protein (Lim et al . 2010). The size and properties of the target proteins are vital in determining the amount of effective domains being displayed on the surface of live cells (Lim et al .…”

Surface Anchoring on Lactococcus lactis by Covalent Isopeptide Bond

“…14,18 Five different types of surface anchoring domains have been described for LAB: transmembrane domains, LPXTG-type domains, lysin motif (LysM) domains, surface layer proteins and lipoprotein anchors. [19][20][21] The most frequently applied surface anchoring domains in prototype LAB, Lactococcus lactis, are the C-terminal part of endogenous AcmA, enabling non-covalent anchoring through peptidoglycan binding LysM repeats, [22][23][24][25] and the LPXTG sequence of M6 protein of Streptococcus pyogenes enabling covalent anchoring. 26,27 Despite these available options, alternative surface display approaches are being sought.…”