Directed evolution of a biterminal bacterial display scaffold enhances the display of diverse peptides.

ABSTRACT: Bacterial cell-surface display systems coupled with quantitative screening methods offer the potential to expand protein engineering capabilities. To more fully exploit this potential, a unique bacterial surface display scaffold was engineered to display peptides more efficiently from the surface exposed C- and N-termini of a circularly permuted outer membrane protein. Using directed evolution, efficient membrane localization of a circularly permuted OmpX (CPX) display scaffold was rescued, thereby improving the presentation of diverse passenger peptides on the cell surface. Random and targeted mutagenesis directed towards linkers joining the native N- and C-termini of OmpX coupled with screening by FACS yielded an enhanced CPX (eCPX) variant which localized to the outer membrane as effici

SUBMITTER: Rice JJ 

PROVIDER: S-EPMC2427320 | biostudies-literature | 2008 Jul

REPOSITORIES: biostudies-literature