Functional modulation and directed assembly of an enzyme through designed non-natural post-translation modification† †Electronic supplementary information (ESI) available: Detailed experimental methods, supplementary Fig. 1 to 11 and supplementary Tables 1 to 3. See DOI: 10.1039/c4sc03900a Click here for additional data file. Click here for additional data file.

Post-translational modification (PTM) modulates and supplements protein functionality. In nature this high precision event requires specific motifs and/or associated modification machinery. To overcome the inherent complexity that hinders PTM's wider use, we have utilized a non-native biocompatible Click chemistry approach to site-specifically modify TEM b-lactamase that adds new functionality. In silico modelling was used to design TEM b-lactamase variants with the non-natural amino acid p-azido-Lphenylalanine (azF) placed at functionally strategic positions permitting residue-specific modification with alkyne adducts by exploiting strain-promoted azide–alkyne cycloaddition. Three designs were implemented so that the modification would: (i) inhibit TEM activity (Y105azF); (ii) restore activity compromised by the initial mutation (P174azF); (iii) facilitate assembly on pristine graphene (W165azF). A dibenzylcyclooctyne (DBCO) with amine functionality was enough to modulate enzymatic activity. Modification of TEM with a DBCO–pyrene adduct had little effect on activity despite the modification site being close to a key catalytic residue but allowed directed assembly of the enzyme on graphene, potentially facilitating the construction of protein-gated carbon transistor systems.