Steady-state experiments are conducted at several angular velocities and at various liquid fractions.
Unless otherwise stated, experiments were generally conducted in triplicates.
Unless otherwise stated, all experiments were conducted at least three times and results are expressed as the mean ± standard deviation.
Unless otherwise stated, all fermentation experiments were conducted in triplicates, and the cultures were incubated at 37 °C and shaking at 150 rpm without pH control.
Solid-state NMR experiments were conducted on a Bruker Avance 800 MHz (18.8 Tesla) spectrometer and a 400 MHz (9.4 Tesla) Bruker Avance spectrometer using 3.2-mm and 4-mm MAS HCN probes, respectively.
As in the iron binding experiments, to keep BH4 in its reduced state binding experiments were conducted in an anaerobic glovebox under an argon atmosphere.
Using optimised solid-state straw culture conditions, experiments were conducted that compared fungal strains in terms of their growth, enzyme activities (cellulases, xylanase and alcohol dehydrogenase) and yield of bioethanol and the undesirable by-products acetic acid and xylitol.
To ensure a constant state of adaptation, all experiments were conducted in a light-tight experimental chamber (2 m wide, 3 m long, and 2.2 m high).
Although the substantial rise in Km indicates that the apparent substrate affinity might be significantly compromised, this is unlikely to limit the pre-steady-state kinetics because the experiments were conducted under saturating substrate concentrations.
To determine the reversal potential for the full open state and the protamine-induced substate, experiments were conducted in the presence of Cs+ and Ca2+ in the cis and trans chambers, respectively.
