A plasmid toolbox for the easy autodisplay of recombinant proteins and its optimization

Furtmann, Christoph; Röhe, Peter; Gesing, Katrin; Kuss, Hanna; Lenz, Florian; Jose, Joachim

Zusammenfassung

Surface display of proteins on bacteria bears several advantages for binding studies, library screening or enzyme inhibitor testing. Here, we present a set of plasmids for autodisplay-based surface expression of recombinant proteins, named Autodisplay-ToolBox (ATB). In this set, crucial parts as required for autodisplay, including promotor, SP, linker and β-barrel can be combined in varying permutations to find the best combination for a certain protein. The plasmids can be applied individually or in library approaches, enabling optimization of surface display in a single step. By such a library approach, the activity of surface-displayed β-glucosidase (β-Gluc) is increased by a factor of 4.9, the activity of a laccase (CotA) by a factor of 4.7 and the binding capacity of the surface-displayed nucleotide binding domain of human HCN2 channel (HCN2-CNBD) by a factor of 10.3. It is shown that the ATB can be used in different strains of E. coli as well as in Pseudomonas putida. The aim is to provide a selection of plasmids, accessible by Addgene, that every scientist can use for their own protein, either individually based on personal preferences, structural features, etc., or as a library, as shown here for three different examples.