Fig. 5: Functional characterization of PolE.
From: Azetidine amino acid biosynthesis by non-haem iron-dependent enzymes
a, Quantitative analysis of PolE time course assays with l-Ile. The reactions were quenched at 30 s, 1 min, 2 min, 4 min, 8 min and 20 min. b, Quantitative analyses of PolEF stepwise assay with l-Ile. PolE assay (500 μl) was performed with 150 μM l-Ile, 15 μM PolE, 100 μM Fe(II), 100 μM BH4, 1 mM ascorbate, 1 mM NADH, 0.5 μM QDPR and ~0.5 mM O2 at room temperature for 40 min. Then 15 μM PolF was added to reaction. The mixtures were incubated for 30 s, 1 min, 2 min, 4 min and 8 min. c, Quantitative analysis of PolEF coupled assays with l-Ile. The assay contains 150 μM l-Ile, 15 μM PolE, 15 μM PolF, 100 μM Fe(II), 100 μM BH4, 1 mM ascorbate, 1 mM NADH, 0.5 μM QDPR and ~0.5 mM O2. The reactions were quenched at 30 s, 1 min, 2 min, 4 min, 8 min and 20 min. d, PA biosynthesis by PolE and PolF. The experiments were performed in triplicate. Error bars represent one standard deviation calculated from the three replicates. Source data for a, b, c are provided.
