Fig. 2: In vitro reconstitution and reaction optimization of the venemycin assembly line.
From: An in vitro platform for engineering and harnessing modular polyketide synthases
a The pH activity profile indicates pH 7.5 is optimal. b In a salt screen, 400 mM potassium phosphate is best. c TCEP is better than DTT as a reductant. d Reactions are fastest at 30–35 °C. e However, the enzymes are not stable at these temperatures - activity after incubation of VemG and VemH at 4–55 °C for 30 min is shown. f Measuring activity at increasing concentrations of VemG (at constant 0.2 µM VemH), enables the determination of the turnover rate and an apparent Kd. g The assembly line accepts 3-hydroxybenzoic acid but is twofold slower. Data are mean ± s.d., n = 3 independent experiments. Source data are provided as a Source Data file.
