Fig. 3: Exploring the dehalogenation activity of SlALR.
a The scheme depicts the possible pathways for the synthesis of an ALD substrate, which can be formed through β-elimination of the F atom or via the action of a racemase upon the addition of DAAO. b Spectrophotometric monitoring of NADH oxidation using FPyr as limiting substrate with different enzyme combinations. The graphs represent mean values and the error bars correspond to standard deviations from three independent experiments. N.B., error bars are smaller than the corresponding mean data point. A.U., arbitrary units. c Time-resolved 19F-NMR monitoring of FAla formation, FPyr consumption and release of free fluoride (F–). The axes are colored according to the chemical species measured in each case. A.U., arbitrary units. d Representative example of the HPLC profiles for assays incubated with either SlALR or EcALR and the blank for the same reaction conditions. The plot shows the region encompassing the characteristic retention time of Pyr. μRIU, refractive index units × 10–6. e Substrate conversion per enantiomer [(S)- and (R)-FAla], determined from the release of Pyr as compared to the initial amount of FAla in the assays. f Catalytic cycles per unit of enzyme for both substrates and different enzyme amounts. In panels e and f, the graphs represent mean values and the error bars correspond to standard deviations from three independent experiments; individual data points are indicated in the plots.
