Table 3 Kinetic parameters of KARI variants determined using NADPH or NADH with saturating concentrations of 2S-AL as substrate.
Origin | β2-αB loop | K M (μM) | k cat (s−1) | k cat /K M (mM−1 s−1) | NADH/NADPH ratio of k cat /K M | Cofactor preference | Ref | ||||
|---|---|---|---|---|---|---|---|---|---|---|---|
NADH | NADPH | NADH | NADPH | NADH | NADPH | ||||||
Sac-KARI | Solfolobus acidocaldarius | LEREGNS | 6.0 ± 0.8 | 0.4 ± 0.2 | 0.056 ± 0.001 | 0.064 ± 0.001 | 9.5 ± 1.3 | 185.8 ± 80.5 | 0.051 | NADPH | This work |
Ia-KARI | Ignisphaer aaggregans | LERQGDS | <1 | <1 | 0.02 | 0.03 | >20 | >25 | ~0.8 | Bispecific | |
Ms-KARI | Metallosphaera sedula | LEREGKS | 24 | 31 | 0.06 | 0.07 | 2.5 | 2.1 | 1.2 | Bispecific | |
Hs-KARI | Hydrogenobaculum sp. | LDDKSPH | 39 | 46 | 0.12 | 0.12 | 3.2 | 2.7 | 1.2 | Bispecific | |
Sw-KARI | Syntrophomonas wolfei | LRKPFDEASEKE | 57 | 44 | 0.28 | 0.22 | 5.0 | 5.0 | 1.0 | Bispecific | |
Tp-KARI | Thermacetogenium phaeum | DIPSSEN | <1 | 40 | 0.46 | 0.25 | >460 | 6.0 | 74 | NADH | |
Ua-KARI | Uncultured Archaeon | ETEILGGNKNPS | 1.1 | 38 | 0.22 | 0.05 | 200 | 1.3 | 152 | NADH | |
Af-KARI | Archaeoglobus fulgidus | LPEWDKAT | 5.0 | 26 | 0.1 | 0.04 | 20 | 1.5 | 13 | NADH | |
Do-KARI | Desulfococcus oleovorans | QLEGDAY | 32 | n.a. | 0.25 | n.a. | 8.0 | n.a. | — | NADH | |
Mr-KARI | Meiothermus ruber | LRPGSRN | 240 | 20 | 1.09 | 0.98 | 4.6 | 54.5 | 0.084 | NADPH | |
So-KARI | Spinacea oleracea (spinach) | LRKGSNS | 101 | 1.7 | 0.95 | 1.33 | 9.4 | 782.4 | 0.012 | NADPH | |
Se-KARI | Slackia exigua | LREGSSS | 45 | 1.0 | 0.41 | 0.8 | 9 | 800 | 0.011 | NADPH | |
Ec-KARI | Escherichia coli | LRKEAIAEKRAS | 1075 | 41 | 0.3 | 3.6 | 0.3 | 88 | 0.003 | NADPH | |
Ec-KARI6E6 | Escherichia coli | LRKESIAEKDAD | 30 | 650 | 2.3 | 0.20 | 74 | 0.40 | 185 | NADH | |
Se-KARIDD | Slackia exigua | LREGDSD | 113 | 880 | 0.97 | 0.10 | 9 | 0.11 | 81.8 | NADH | |
Se-KARIDDV | Slackia exigua | LREGDSD | 47 | >1000 | 1.01 | 0.25 | 22 | 0.25 | 88 | NADH | |
