Fig. 3
From: On-cell catalysis by surface engineering of live cells with an artificial metalloenzyme
Development of an artificial allylic deallylase for the activation of proCoumarin 4. a Cleavage of protected amines for prodrug activation. b Uncaging of allyl-carbamate-protected proCoumarin 4 as our model reaction. c Development of the artificial allylic deallylase 10 based on ruthenium complex 7 and biotinylated ligand 8. d In vitro screening of artificial allylic deallylases for the deprotection of proCoumarin 4. Reaction conditions: proCoumarin 4 (500 µM), ruthenium cofactor 9 (5 µM in DMF), purified streptavidin isoforms (10 µM, free biotin-binding sites), PBS (1×, pH 7.4), 0.5% DMF, 25 °C, 250 rpm shaking, 4 h. Fluorescence of product 6 was determined in a plate reader at λex. = 395 nm and λem. = 460 nm from triplicate reactions. Conversions were calculated by comparison with a standard curve of coumarin 6 (see Supplementary Figure 7 for details). Data are represented as mean ± SD (n = 3)
