Table 1.

Kinetic parameters for the EcDHFR‐catalyzed hydride transfer reactions at 25 °C.

Enzymes	E A [kcal mol−1]	ΔE A [kcal mol−1]	ΔS ≠ [cal mol−1 K−1]	ΔΔS ≠ [cal mol−1 K−1]	ΔH ≠ [kcal mol−1]	ΔΔH ≠ [kcal mol−1]	ΔG ≠ [kcal mol−1]
Light WT EcDHFR[a]	7.3±0.2		−26±1		6.7±0.3		14.4±1.5
Light EcDHFR‐A29C	7.3±0.3		−25±1		6.8±0.3		14.3±1.5
Heavy WT EcDHFR[a]	6.0±0.3	1.3±0.4	−30±2	4±2	5.4±0.6	1.3±0.7	14.4±2.5
Light, chemically ligated EcDHFR	8.3±0.3		−22±1		7.8±0.3		14.3±1.3
NT‐EcDHFR	8.4±0.2	−0.1±0.4	−22±2	0±2	7.8±0.6	0.0±0.7	14.3±1.9
CT‐EcDHFR	7.0±0.4	1.3±0.5	−27±2	5±2	6.5±0.8	1.3±0.9	14.3±3.5

[a] Data from Ref. 1g. WT=wild type.