. Author manuscript; available in PMC: 2012 Nov 1.

Published in final edited form as: Bioorg Med Chem. 2011 Aug 30;19(21):6447–6453. doi: 10.1016/j.bmc.2011.08.056

Table 2.

Kinetic parameters for KHO cleavage catalyzed by selected TM KDPG aldolase mutants.^*

KDPG aldolase	k_cat (s⁻¹)	K_M (mM)	k_cat/K_M (M⁻¹s⁻¹)	relative k_cat/K_M
wt-TMA	0.40 ± 0.05	200 ± 30	2.2 ± 0.3	1
TM 1	0.07 ± 0.01	9 ± 2	9.3 ± 1.8	4
TM 2	0.5 ± 0.1	90 ± 20	5.5 ± 1.0	2.5
TM 3	0.14 ± 0.10	20 ± 14	7 ± 4	3
TM 4	0.36 ± 0.02	23 ± 4	16 ± 2	7
TM 5	0.75 ± 0.10	14 ± 5	54 ± 10	25
TM 6	0.13 ± 0.02	8 ± 3	17 ± 6	8
TM 8	1.2 ± 0.3	49 ± 19	25 ± 5	11

Initial velocity for pyruvate formation was measured using a lactate dehydrogenase coupled assay at 34°C. Steady state kinetic parameters were determined by fitting of the Michaelis-Menten equation to the initial velocity as a function of the substrate concentration.