Table 3.

Steady-state kinetic analysis of mutant AldC. The protein and location of the mutation are indicated. Enzyme assays were performed as described under “Experimental procedures.” Average values ± S.D. (n = 3) are shown.

Protein	Ligand site	Octanal			NAD+
		Kcat	Km	Kcat/Km	Kcat	Km	Kcat/Km
		min−1	µm	m−1 s−1	min−1	µm	m−1 s−1
WT		67.3 ± 1.4	1,210 ± 93	927	30.9 ± 0.4	490 ± 25	1,050
W158A	NAD(H)	2.6 ± 0.1	340 ± 44	127	5.4 ± 0.1	643 ± 57	140
W158H	NAD(H)	13.7 ± 0.5	804 ± 117	284	6.9 ± 0.5	273 ± 38	406
E185A	NAD(H)	4.0 ± 0.2	4,380 ± 419	15	5.2 ± 0.1	1,900 ± 120	46
E185Q	NAD(H)	79.5 ± 2.5	1,620 ± 150	818	15.0 ± 0.2	236 ± 14	1,060
T234A	NAD(H)	21.6 ± 1.2	1,520 ± 300	237	6.8 ± 0.7	594 ± 217	191
S236A	NAD(H)	7.0 ± 0.4	5,190 ± 689	22	6.4 ± 0.2	1,420 ± 143	75
S236T	NAD(H)	3.0 ± 0.2	527 ± 187	95	7.3 ± 0.4	2,200 ± 294	55
E257Q	NAD(H)	9.8 ± 1.0	530 ± 258	308	15.7 ± 4.5	287 ± 170	912
E391D	NAD(H)	25.3 ± 2.4	2,730 ± 743	154	10.3 ± 3.0	248 ± 183	692
M114A	Octanal	22.0 ± 1.0	1,220 ± 218	301	11.7 ± 0.5	543 ± 80	360
L118A	Octanal	17.0 ± 1.7	1,080 ± 432	262	11.0 ± 0.4	298 ± 34	615
W160Y	Octanal	63.7 ± 4.5	2,160 ± 472	491	6.1 ± 0.6	516 ± 26	197
Y163A	Octanal	70.2 ± 8.7	6,540 ± 1,580	179	18.8 ± 5.3	244 ± 162	1,280
Q164A	Octanal	76.3 ± 17.6	4,110 ± 2,250	309	29.2 ± 5.3	416 ± 213	1,170
Q164N	Octanal	66.0 ± 7.9	2,270 ± 819	485	24.8 ± 1.5	258 ± 38	1,600
R285A	Octanal	16.5 ± 0.5	1,100 ± 128	250	11.8 ± 0.5	434 ± 66	453
S290A	Octanal	2.8 ± 0.1	830 ± 180	56	7.5 ± 0.2	1,220 ± 81	102
W450A	Octanal	9.8 ± 1.0	220 ± 90	742	5.1 ± 0.3	805 ± 167	106
F456W	Octanal	140 ± 33	5,420 ± 2,680	431	2.9 ± 0.1	622 ± 5	78
Y468A	Octanal	8.4 ± 0.6	2,520 ± 548	56	8.6 ± 0.3	743 ± 81	193