TABLE 3.
Substrate | Benzaldehydea |
N1-Methylnicotinamida |
||||
---|---|---|---|---|---|---|
kcat | Km | kcat/Km | kcatb | Km | kcat/Km | |
min−1 | μm | μm/min | min−1 | μm | μm/min | |
mAOX3-WT | 41.9 ± 0.8 | 2.5 ± 0.2 | 16.76 | 14.7 ± 0.1 | 128.5 ± 5.8 | 0.115 |
E1266Q | 10.2 ± 0.5 | 86.3 ± 11.3 | 0.12 | ND | ND | |
A807V | 41.0 ± 0.3 | 2.9 ± 0.2 | 14.14 | 11.2 ± 0.1 | 123.4 ± 5.4 | 0.091 |
Y885M | 57.4 ± 0.8 | 6.3 ± 0.3 | 9.11 | 23.4 ± 0.4 | 82.7 ± 6.6 | 0.283 |
A807V/Y885M | 43.6 ± 1.1 | 4.6 ± 0.4 | 9.48 | 27.0 ± 0.2 | 35.9 ± 1.4 | 0.752 |
K889H | 19.2 ± 0.5 | 2.3 ± 0.3 | 8.20 | 3.1 ± 0.1 | 25.5 ± 4.0 | 0.120 |
Substrate | Phenanthridineb |
Phthalazinea |
||||
---|---|---|---|---|---|---|
kcat | Km | kcat/Km | kcat | Km | kcat/Km | |
min−1 | μm | μm/min | min−1 | μm | μm/min | |
mAOX3-WT | 51.7 ± 0.6 | 32.3 ± 1.4 | 1.60 | 41.1 ± 1.0 | 1.4 ± 0.2 | 29.36 |
E1266Q | ND | ND | ND | ND | ||
A807V | 315.0 ± 5.2 | 149.5 ± 5.8 | 2.11 | 41.1 ± 1.5 | 2.7 ± 0.4 | 15.22 |
Y885M | 266.9 ± 66.5 | 33.4 ± 11.6 | 7.99 | 48.9 ± 1.2 | 3.2 ± 0.2 | 15.28 |
A807V/Y885M | 218.0 ± 17.1 | 11.8 ± 1.7 | 18.47 | 41.4 ± 1.8 | 2.9 ± 0.3 | 14.28 |
K889H | 79.9 ± 0.7 | 16.1 ± 0.6 | 4.97 | 11.8 ± 0.3 | 1.1 ± 0.2 | 10.53 |
a As terminal electron acceptor 100 μm DCPIP were used.
b As terminal electron acceptor molecular oxygen in air saturated buffer was used.