To obtain sufficient2H-2, a 5 mL-total-volume (D2O:H2O = 9:1) large scale enzymatic reactions were performed. The solution contained 50 mM phosphate buffer (pH 7.0), 1.2 mM3and 40 μM AvmM. After incubation at 30 °C for 2 h, 5 mL acetonitrile was added to qu...
Electrostatic interactions were treated by using the particle–mesh Ewald (PME) method with a grid spacing of 0.16 Å and a spline interpolation of order 4 [73]. The cross-interactions for Lennard–Jones terms were calculated using the Lorentz–Berthelot mixing rules and intramolecular interactions...