Table 1.

Enzyme kinetics of native PtmA2 and designed mutants^a

Protein	Substrate	Km (μM)	Vmax (μmol s−1 × 10−2)	kcat (s−1)	kcat/Km (s−1 M−1)
Native	6	1.5 ± 0.2	21.5 ± 0.4	22	1.5 × 107
	CoAb	6.2 ± 0.6	9.7 ± 0.3	20	3.2 × 106
	9	1.0 ± 0.1	20.6 ± 0.3	21	2.1 × 107
	CoAc	2.5 ± 0.4	9.3 ± 0.3	19	7.6 × 106
E416A	6	0.42 ± 0.11	10.7 ± 0.3	0.21	5.0 × 105
	CoAb	7.8 ± 1.2	8.9 ± 0.4	0.18	2.3 × 104
A497K	6	1.6 ± 0.2	8.9 ± 0.4	26	1.6 × 107
	CoAb	6.8 ± 1.0	7.9 ± 0.4	32	4.7 × 106
P-loop	6	0.58 ± 0.12	5.1 ± 0.1	10	1.7 × 107
	CoAb	70 ± 8	14.8 ± 0.4	7.4	1.1 × 105
A497K/P-loop	6	0.71± 0.11	14.3 ± 0.3	14	2.0 × 107
	CoAb	69 ± 11	9.9 ± 0.4	10	1.4 × 105

^aValues are the means of three independent assays and reported with standard deviations.

^bKinetics determined with saturating concentration (100 μM) of 6.

^cKinetics determined with saturating concentration (100 μM) of 9.