TABLE 2.

Kinetic parameters of EfAmy and site-directed variants at 5, 25, and 35°C

Enzyme or mutation(s)	Temp (°C)	kcat (s−1)	Km (g · liter−1)	kcat/Km (s−1 · g−1 · liter)
EfAmy	5	718.55 ± 31.91	3.05 ± 0.08	235.59 ± 16.65
	25	1466.42 ± 46.1	3.31 ± 0.16	443.03 ± 35.43
	35	982.5 ± 33.18	4.43 ± 0.19	221.78 ± 17.03
EcAmy	5	377.53 ± 19.33	1.31 ± 0.04	288.19 ± 23.58
	25	618.91 ± 23.92	1.36 ± 0.03	455.08 ± 27.64
	35	879.76 ± 27.61	1.42 ± 0.07	619.55 ± 50.11
E166P	5	694.31 ± 26.29	2.91 ± 0.06	238.59 ± 13.96
	25	1452.39 ± 37.49	3.21 ± 0.07	452.46 ± 21.56
	35	1033,78 ± 38.91	4.01 ± 0.12	262.79 ± 17.58
S185P	5	677.62 ± 26.41	2.86 ± 0.13	236.93 ± 20.05
	25	1407.58 ± 33.96	3.04 ± 0.11	463.02 ± 27.96
	35	1162.17 ± 31.42	3.89 ± 0.08	298.76 ± 14.23
T350P	5	704.62 ± 23.19	2.99 ± 0.09	235.65 ± 14.86
	25	1458.29 ± 32.48	3.23 ± 0.08	451.48 ± 21.25
	35	999.76 ± 29.35	4.18 ± 0.18	239.18 ± 17.35
V212T	5	681.49 ± 28.67	2.36 ± 0.07	288.77 ± 20.73
	25	1392.11 ± 42.49	2.67 ± 0.11	521.39 ± 37.46
	35	1002.34 ± 34.72	2.81 ± 0.16	356.71 ± 32.77
V232T	5	693.89 ± 27.33	2.42 ± 0.15	286.73 ± 29.18
	25	1409.63 ± 37.54	2.66 ± 0.09	529.94 ± 32.08
	35	1011.87 ± 39.69	2.81 ± 0.12	360.1 ± 29.56
V212T/V232T	5	643.28 ± 31.42	1.76 ± 0.13	365.51 ± 45.1
	25	1314.08 ± 41.66	1.89 ± 0.09	695.28 ± 55.28
	35	1039.36 ± 34.51	2.01 ± 0.11	517.09 ± 45.61
E166P/S185P/T350P	5	632.57 ± 21.65	2.69 ± 0.11	235.16 ± 17.69
	25	1297.12 ± 41.6	2.89 ± 0.13	448.83 ± 34.65
	35	1304 ± 38.85	3.76 ± 0.18	346.81 ± 27.01
E166P/S185P/T350P/V212T/V232T	5	483.75 ± 23.28	1.53 ± 0.05	316.18 ± 25.58
	25	983.16 ± 29.61	1.59 ± 0.07	618.34 ± 45.93
	35	1332.03 ± 38.37	1.67 ± 0.11	797.6 ± 75.84