Yeast surface display, also known as yeast display, is a protein engineering technique that uses the
expression of recombinant proteins of interest, fused with native cell wall proteins of yeast. Ply511
endolysin is a cell wall hydrolase with N-acetylmuramoyl-L-alanine amidase activity.
Ply511 is naturally produced by the Listeria phage A511 to lyse bacterial cells and has been
proposed as an alternative to antibiotics.
Listeria monocytogenes is a Gram-positive pathogen causative of human infections, resulting in
febrile gastroenteritis, perinatal infection, and central nervous system infections. These infections
are frequently acquired through the ingestion of contaminated food.
In our study, we employed CRISPR-Cas9 to genetically modify Saccharomyces cerevisiae to
display the listeria endolysin Ply511 on its surface. Flow cytometer analyses confirmed the
expression of the genetic construction integrated in the recombinant yeast. Also, the enzymatic
peptidoglycan-degrading activity of the engineered yeast was confirmed by using heat-killed L.
monocytogenes cells. In killing assays, the recombinant yeast did not show CFU reduction of
Listeria in comparison with the wild-type. These results suggest that yeast display of endolysins
needs to be improved to be used as an antimicrobial strategy in the context of engineered
probiotics, to assure that bacterial killing is achieved.info:eu-repo/semantics/publishedVersio
