Fig. 1: Core and Shell variants of Kemp eliminases. | Nature Communications

Fig. 1: Core and Shell variants of Kemp eliminases.

From: Distal mutations enhance catalysis in designed enzymes by facilitating substrate binding and product release

Fig. 1

a Kemp eliminases catalyze the concerted deprotonation and ring opening of benzisoxazoles using an Asp, Glu, or His catalytic base. b Core and Shell variants from the HG3, 1A53, and KE70 families contain either active-site (purple) or distal (yellow) mutations found by directed evolution, respectively. Mutations are listed on Supplementary Table 2. Catalytic residues are shown as gray spheres and the bound transition-state analogue 6-nitrobenzotriazole (6NBT) is shown as orange sticks. c Michaelis–Menten plots of normalized initial rates as a function of substrate concentration are shown, with smaller graphs providing close-up views of the Designed and Shell variants. Data represent the average of six or nine individual replicate measurements from two or three independent protein batches, with error bars indicating the SEM (mean ± SEM, n = 6 or 9). d 6NBT tautomerizes in water, which allows it to bind to enzymes that react with either 5- or 6-nitrobenzisoxazole. Source data are provided as a Source data file.

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