Investigation of Listeria Phage Endolysin Cell-wall Binding Domain (CBD) Surface Display in Escherichia coli

Abstract

Cell surface display of target proteins has been widely used in biotechnology and industry. In this study, cell-wall binding domain (CBD) from Listeria phage endolysin A500 was fused with anchoring domains of YadA and OmpA, respectively, in Escherichia coli, aiming at binding of E. coli cells to Listeria cells. The fusion proteins were expressed after induction and their surface localizations were verified by Western blot. CBD-YadA fusion was displayed on the cell surface, however, was toxic to E. coli. OmpA-CBD fusion was translocated to the outer layer of the cell membrane but with compromised availability on the cell surface. Therefore, functional surface display of CBD in E. coli requires another anchor for fusion strategy.