Table 3.

Changes in secondary structure, ligand free folding energy, ligand free energy of binding and ligand affinity in fifteen double mutants of A. thaliana ent-CPS

Nr	Mutation	ΔΔGFOLD (kcal/mol)	Secondary structure change and accuracy of prediction (%)	ΔΔGBIND (kcal/mol)	KA (nM)
1	T114F/D336L	1.83	None, 100%	− 0.64	97.61
2	T114F/D377L	− 0.89	None, 100%	− 0.60	106.56
3	T114F/G422L	− 2.87	None, 100%	− 0.18	216.51
4	T114F/S597W	− 1.62	None, 100%	− 0.55	116.14
5	T114F/K778F	0.30	None, 100%	− 0.02	282.68
6	D336L/D377L	− 1.94	None, 100%	− 0.48	129.39
7	D336L/G422L	− 1.93	None, 100%	− 0.10	249.07
8	D336L/S597W	− 1.84	None, 100%	− 0.74	84.56
9	D336L/K778F	− 1.67	None, 100%	− 0.79	77.46
10	D377L/G422L	− 3.24	None, 100%	− 0.72	87.03
11	D377L/S597W	− 2.43	None, 100%	− 0.79	77.33
12	D377L/K778F	− 2.43	None, 100%	− 0.79	78.12
13	G422L/S597 W	− 3.39	None, 100%	− 0.04	284.59
14	G422L/K778F	− 3.39	None, 100%	− 0.47	132.71
15	S597W/K778F	− 2.35	None, 100%	− 0.13	234.78

Seven selected double mutants that passed the presented quality test are marked in bold