TABLE 1.

Kinetic parameters of AGT‐Ma, AGT‐Mi, and the variants chosen from the AGT library

Enzyme	Substrate	Cosubstrate	k cat (s−1)	K M (mM)	k cat/K M (s−1 mM−1)
AGT‐Ma a	l‐alanine	Glyoxylate	45 ± 2	31 ± 4	1.4 ± 0.2
	Glyoxylate	l‐alanine	45 ± 3	0.23 ± 0.05	196 ± 44
AGT‐Mi b	l‐alanine	Glyoxylate	33 ± 5	28 ± 2	1.2 ± 0.2
	Glyoxylate	l‐alanine	37 ± 1	0.22 ± 0.01	168 ± 8
S99A	l‐alanine	Glyoxylate	36 ± 2	29 ± 4	1.2 ± 0.18
	Glyoxylate	l‐alanine	34 ± 3	0.31 ± 0.05	109 ± 20
P125Q	l‐alanine	Glyoxylate	43 ± 5	55 ± 8	0.78 ± 0.14
	Glyoxylate	l‐alanine	35 ± 2	0.22 ± 0.03	159 ± 22
T127A	l‐alanine	Glyoxylate	42 ± 3	33 ± 4	1.27 ± 0.17
	Glyoxylate	l‐alanine	34 ± 4	0.16 ± 0.03	212 ± 44
Q145R	l‐alanine	Glyoxylate	47 ± 4	41 ± 5	1.15 ± 0.17
	Glyoxylate	l‐alanine	38 ± 4	0.21 ± 0.04	181 ± 32
K177Q	l‐alanine	Glyoxylate	37 ± 3	35 ± 5	1.05 ± 0.17
	Glyoxylate	l‐alanine	36 ± 4	0.27 ± 0.06	133 ± 33
E138D/K177Q	l‐alanine	Glyoxylate	44 ± 4	53 ± 7	0.83 ± 0.13
	Glyoxylate	l‐alanine	39 ± 3	0.27 ± 0.04	145 ± 24

Note: Experimental details are given in Section 4. Experiments have been performed in triplicate.

^a From Ref. 10.

^b From Ref. 83.