Table 1 Biochemical characterization of extant (FSD-Noca and FSD-Cryar) and ancestral (AncD1) recombinant enzymes.
From: Reconstructing the evolutionary history of F420-dependent dehydrogenases
Substrates | FGD-Noca (FSD-Noca) | FGD-Cryar (FSD-Cryar) | AncD1 | |||
|---|---|---|---|---|---|---|
kcat [s−1] | KM [mM] | kcat [s−1] | KM [mM] | kcat [s−1] | KM [mM] | |
d-glucose-6-phosphate | 4.1 ± 0.24 | 0.94 ± 0.19 | 6.2 ± 0.82 | 0.9 ± 0.4 | 0.9 ± 0.05 | 1 ± 0.19 |
d-fructose-6-phosphate | 2.1 ± 0.2 | 4.5 ± 1.1 | 3.6 ± 0.41 | 6.1 ± 1.7 | 0.3 ± 0.05 | 45 ± 15 |
d-mannose-6-phosphate | 2.6 ± 0.15 | 16.4 ± 2.5 | 0.23 ± 0.03 | 7 ± 2.7 | 0.04 ± 0.01 | 7.2 ± 4.7 |
d-glucose | — | >500 | — | >500 | — | >500 |
Substrates [400 mM] | k obs [s −1 ] | k obs [s −1 ] | k obs [s −1 ] | |||
d-fructose | — | — | — | |||
d-mannose | 0.02 ± 0.01 | — | 0.1 ± 0.03 | |||
d-xylose | 0.11 ± 0.05 | 0.13 ± 0.1 | — | |||
Binding affinity | K D [µM] | KD [µM] | KD [µM] | |||
F420 | 0.09 ± 0.02 | 2.2 ± 0.7 | 1.6 ± 0.1 | |||
FMN | >150 | >150 | >150 | |||
Activity features | ||||||
pH | 6.5 | 7.5 | 6.5 | |||
Tm [°C] | 45.5 | 43 | 53 | |||
