Fig. 10
From: Combinatorial assembly and optimisation of designer cellulosomes: a galactomannan case study
VersaTile platform for the optimisation of designer cellulosomes. Step 1: pick a substrate and select catalytically active tiles from the docking enzyme tile repository. Here, GM was chosen and five GM-degrading enzymes were selected. Step 2: docking enzyme optimisation: VersaTile allows the quick and easy construction of docking enzymes composed of enzyme, dockerin and/or linker tiles. Here, 79 different docking enzymes were designed. Activity assays allowed us to select three optimised docking enzyme variants to incorporate in the final complex. Step 3: scaffoldin optimisation: VersaTile allows the quick and easy construction of scaffoldins composed of five cohesins and/or CBM tiles. Here, 72 different scaffoldins were designed. Activity analysis of complexes composed of these scaffoldins and the selected docking enzymes allowed us to construct an optimised designer cellulosome