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. 2020 Dec 3;48(22):12845–12857. doi: 10.1093/nar/gkaa1147

Figure 3.

Figure 3.

In vitro photoreduction and anaerobic purification of NewPHLs. (A) Photoreduction of DsNewPHL with 50 mm DTT one day after purification starting with FADHo. All photoreduction experiments were carried out with blue light with a maximum emission wavelength of 455 nm (ILS – Intelligent LED Solutions) and a light intensity of 5.9 mW·cm−2 (224 μmol·m−2·s−1) at 4 cm distance. (B) Photoreduction of DsNewPHL after in vitro oxidation with potassium ferricyanide to FADox. (C) Absorption spectra of DsNewPHL directly after IMAC purification under strictly anaerobic conditions. Difference spectra of aerobic oxidation after anaerobic purification for 300 min, measured with difference between time points T30 min and T300 min is shown within the inlay on the upper right side. (D) Photoreduction of MmNewPHL with 50 mm DTT one day after purification, starting with a mixture of different FAD redox states. (E) The photoreduction of MmNewPHL starting with FADox. (F) The absorption spectrum of MmNewPHL after IMAC under strictly anaerobic conditions.