Fig. 1

Flowchart of the key steps in the computational redesign of the active site of TLL. a Architecture of the substrate binding sites in TLL. The yellow surface represents the residues located in the lid of the open conformation. The active site residues located in the nucleophilic (alcohol-binding) and electrophilic (acid-binding) pockets are represented in chartreuse and salmon, respectively. b Multipoint mutants in two rounds of redesign are exhaustively enumerated using an automated web-accessible server, FuncLib (https://funclib.weizmann.ac.il/bin/steps), and represented using WebLogo (https://weblogo.threeplusone.com/) to generate a sequence logo. The height of each letter represents the substitution probability of the amino-acid residue [35]. c The designs were ranked by energy, and designs with low energy were selected for experimental construction. Mutations in the same designed position are shown in consistent colors