1. Redox potential in flavin FAD + e - → FAD - Calculating accurate redox potentials in enzymes with a combined QM/MM free energy perturbation approach – J. Theor. Comp. Chem.

3. B3LYP/631 + G(d,p) SCC-DFTB (semi-empirical DFT) AM1 -44.3 kcal/mol = -1.92 eV -37.3 kcal/mol = - 1.62 eV -63.5 kcal/mol = -2.76 eV

4. *** use eq. coord. of neutral state for charged state energy calculation (not optimize) Energy calculation using GAMESS optimizeSTO-3N31-6*** Oxidized (FAD) -860.467 Hartree -871.347 Hartree Reduced (FAD - ) -860.419 Hartree Not yet (-871.40??) -871.397 Hartree ΔE (eV)-1.31eV(?) (1.63 eV?) (ΔE -1.92 eV) 1.36 eV (31.2 kcal/mol) Rms (Å)0.0453

5. -|V 0 | = -45 kcal/mol, λ 0 =1/2kq 0 2 =45kcal/mol (-37.6?) ( 6.4?) ∂G/∂λ (kcal/mol) Ref. GAMESS (N31-6) λ=0 ~ 0-31.2 -|V 0 |+ λ 0 λ=0.5 ~ -45-37.6 (?) (not yet) -|V 0 | λ=1 ~ -90 -|V 0 |- λ 0