Temperature dependency of CYP152BSβ using ABTS (a) and guaiacol (b) as non-natural substrates.
Product conversion was measured photometrically at 405 nm or 470 nm, respectively. Activity assays were performed at different temperatures using a UV/VIS spectrometer with an integrated Peltier element. All assay components were incubated at the respective temperatures for 5 min prior to starting the enzymatic reaction. Product formation was calculated using Lambert-Beer law (εABTS 36.8 mmol l-1 cm-1; εguaiacol 26.6 mmol l-1 cm-1). Specific activities are given in mmol l-1 min-1 mg-1 and were calculated based on the initial velocities of the reactions. Means and standard deviations reflect three experiments.