. 2003 Mar 7;100(6):3143–3148. doi: 10.1073/pnas.0635924100

Table 1.

Steady-state kinetic parameters for wild-type and mutant aldolases

Enzyme	FBP cleavage			TBP cleavage			k_cat/K_m (FBP)/ k_cat/K_m (TBP)
Enzyme	k_cat, min⁻¹	K_m, mM	k_cat/K_m, min⁻¹⋅mM	k_cat, min⁻¹	K_m, mM	k_cat/K_m, min⁻¹⋅mM	k_cat/K_m (FBP)/ k_cat/K_m (TBP)
FBPA	630 ± 4	0.17 ± 0.003	3,700	0.9 ± 0.08	0.35 ± 0.06	2.6	1,423
TBPA (AgaY)	4.1 ± 0.35	1.3 ± 0.32	3.2	280 ± 8	0.26 ± 0.03	1,080	0.003
1-20F8 (D104G)	3 ± 0.21	0.26 ± 0.05	12	29 ± 2.4	0.15 ± 0.03	193	0.06
1-20H6 (S106G)	9.6 ± 0.45	0.37 ± 0.05	26	380 ± 8	0.05 ± 0.004	7,600	0.0034
1-24H12 (H26Y)	24 ± 0.96	0.27 ± 0.03	89	38 ± 3.4	0.018 ± 0.005	2,533	0.04
2-5C1 (H26Y/D104G/V121A)	39 ± 0.98	0.16 ± 0.01	244	16 ± 0.36	0.013 ± 0.001	1,231	0.20
3-23B5 (H26Y/D104G/ V121A/P256L)	36 ± 0.81	0.16 ± 0.01	230	13 ± 0.14	0.016 ± 0.001	800	0.3

The steady-state kinetic parameters of the wild-type and mutant enzymes for FBP and TBP cleavage were measured by using a coupled enzyme assay (29). Kinetic parameters (±standard error of the fit) were determined by fitting the data to the Michaelis–Menten equation by using the program kaleidagraph (Abelbeck Software, Reading, PA). FBPA, FBP aldolase; TBPA, TBP aldolase.