| X-RAY DIFFRACTION |
4.6 |
293.15 |
Prior to crystallisation, fresh sample was centrifuged at 13 000 g for 3 min to remove macro-aggregates and dust. The sample was crystallised in an anaerobic chamber filled with a N2/H2 (97:3%) atmosphere, at 20 degrees Celsius. The crystallisation was done in 96-Well MRC 2-Drop polystyrene plates (SWISSCI) containing 90 uL of crystallisation solution in the reservoir. 0.7 uL of enzyme at 6.5 mg/ml + 1 mM FAD was mixed with 0.7 uL of the crystallisation solution. The crystallisation solution contained 45 % w/v pentaerythritol propoxylate (5/4 PO/OH), 100 mM sodium acetate pH 4.6, and 400 mM potassium chloride. Prior to flash-freezing in liquid nitrogen, crystals were soaked in the crystallisation solution supplemented with 50 mM sodium nitrite for 5 min. |
