Fig. 2: Evaluating the activity of StDAPDH and VpALD in vitro. | Communications Chemistry

Fig. 2: Evaluating the activity of StDAPDH and VpALD in vitro.

From: Enzymatic synthesis of mono- and trifluorinated alanine enantiomers expands the scope of fluorine biocatalysis

Fig. 2

a Kinetic data for the two enzymes assayed against Pyr, FPyr and F3Pyr, with fitting to the canonical Michaelis-Menten equation. Kinetics were determined from three independent experiments; individual slope values are indicated in the plots. b Time-resolved 19F-NMR monitoring of FAla and F3Ala production. The plots show the integration of the product peak over time for both VpALD and StDAPDH; due to the disparity on the levels of F3Ala formation with the later enzyme, a secondary y-axis is shown (identified with the same color code as per the enzyme assayed). A.U., arbitrary units. c LC-MS spectra indicating the peaks with the m/z signal predicted for FAla and F3Ala. The results shown for VpALD and StDAPDH are compared to the corresponding blank assay with no added enzyme. A.U., arbitrary units. d Representative 19F-NMR spectra for the assays producing FAla and F3Ala using the corresponding enzymes and substrates. The plots display a zoom-in of the chemical shifts predicted for each of the fluorinated products. The spectrum for F3Ala formation by StDAPDH was acquired using 4 times more scans than for VpALD in order to increase the signal intensity.

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