Fig. 3: Results of engineering and optimizing whole-cell catalysis conditions for PcPAL in β-MeCA amination. | Nature Communications

Fig. 3: Results of engineering and optimizing whole-cell catalysis conditions for PcPAL in β-MeCA amination.

From: Direct asymmetric synthesis of β-branched aromatic α-amino acids using engineered phenylalanine ammonia lyases

Fig. 3

a Key bulky residues around the substrate β-MeCA modeled in PcPAL-WT. The active site illustrations are based on the crystal structure of PcPAL (Protein Data Bank ID: 6HQF). The substrate and MIO group are highlighted in orange and gray, respectively. b Enzyme activity of PcPAL mutants towards β-MeCA. Reaction conditions: 100 µg purified enzyme, 1 mM β-MeCA, total 200 µl in NH4OH buffer (pH 10.0), 30 °C, 450 rpm for 24 h. After that the reaction was stopped by adding HCl and centrifuged for further HPLC analysis. c Time curves with different substrate concentrations. Three parallel samples were collected from each reaction for HPLC analysis. Product concentrations were calculated using a standard curve and the peak area of the product. Error bars (SD) were derived from triplicate experiments and created using GraphPad Prism 8. Data are presented as mean values ± SD. Source data are provided in Source Data 1. β-MeCA: β-methyl cinnamic acid; MIO: 3,5-dihydro-5-methylidene-4H-imidazol-4-one.

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