Figure 4.

Hydroxylated C12:0 as substrate for P450 BM3‐catalyzed oxy‐functionalization. Top panel: gas chromatography traces from the reaction before and after incubation for 17 hr. The horizontal axis shows time (min) and the vertical axis flame ionization detection signal in arbitrary units. The reaction (10 ml) was composed of 10 µM his_BM3, 9.5 U/ml commercial GDH, 20 mM C12:0‐OHs (41.6 mg), 5% (by volume) DMSO, 200 mM glucose, 500 µM NADP⁺, 5 mg/ml catalase, and 50 potassium phosphate buffer (pH 7.5). The reaction was operated at 5 ml/min O₂ gassing, 250 rpm magnetic stirring, and automated pH control (pH 7.2). Bottom panel: Correlation between overoxidized product formed and decrease in coupling efficiency. Data are from the reaction in Figure 2 (bottom panel). DMSO, dimethyl sulfoxide [Color figure can be viewed at wileyonlinelibrary.com]