Abstract

Our project aims to improve the loading capacity of yeast surface display system by displaying microtubules and flagellar filaments, respectively, onto the yeast surface. To start with, By using the agglutinin system, we anchored subunits of fibrous polymers which are represented here by microtubulin subunits or FilC, on the cell wall of yeast. Since the particular polymers self-assembly can take place outside cell membrane, we provided the engineered yeast with an environment rich of pre-secreted polymers’ subunits. In this way, an extracellular tridimensional display system could be formed, which can enlarge the loading capacity by roughly two orders of magnitude. Moreover, we remodified the structure of FliC, substituting its D3 domain by the particular enzyme. By means of displaying and assembling specific combinations of FilC-enzymes, the co-display ability can be greatly enhanced. As a result, a highly efficient, whole-cell biocatalyst system would be established in the meantime.