Fig. 2: Identification of historical mutations responsible for the evolution of stereoselectivity. | Nature Communications

Fig. 2: Identification of historical mutations responsible for the evolution of stereoselectivity.

From: Evolutionary insights into the stereoselectivity of imine reductases based on ancestral sequence reconstruction

Fig. 2

a Simplified phylogenetic tree showing the distribution of ancestral and eight extant sequences (left), and six coevolution mutations identified via ibis2Analyzer (right). Light purple and yellow represent the S- and R-stereoselectivity, respectively. The extant enzymes originate from Amycolatopsis azurea (AaIR), Amycolatopsis decaplanina (AdIR), Streptomyces viridochromogenes (SvIR), Saccharothrix espanaensis (SeIR), Streptomyces turgidiscabies (StIR), Streptomyces sp. GF3587 (RIR). The ee of all extant IREDs cited from our previous study20. b Enantiomeric excess (ee) of 2a produced by the imine reduction using N559, N559-M6, N560 and N560-M6. N559-M6, a variant of N559 with mutations L19M, C67S, T94S, A119G, I120V, and D170Y. N560-M6, a variant of N560 with mutations M19L, S67C, S94T, G119A, V120I, and Y170D. c Comparison of the stereoselectivity of N559 and its variant N559-M6 toward other substrates 1b−1f. Numbers above histograms represent ee of the product. The ee of product from each enzyme was calculated from the average of triplicate experiments (n = 3). The data are presented as mean values ± SEM. Light purple and yellow bars represent the percentage of S- and R-enantiomers produced by IREDs, respectively.

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