Protein display on the Yarrowia lipolytica yeast cell surface using the cell wall protein YlPir1

Abstract

A cell surface display system has been developed in the yeast Yarrowia lipolytica using the cell wall protein YlPir1. The red fluorescent protein TurboFP635 was used as a model protein. The expression plasmid in which the YlPIR1 gene without stop-codon was fused in-frame with the nucleotide sequence encoding TurboFP635 was constructed. The thus obtained recombinant protein complex consists of the YlPir1 the C-terminus of which is fused to the N-terminus of red fluorescent protein. Cell surface display of TurboFP635 was confirmed by fluorescent microscopy. This N-terminal system of protein incorporation in the cell wall could be efficiently applied to the cell surface display of enzymes with active sites located near the C-terminus, for example, lipases.