Table 3.
H2-driven NAD+ reduction activity of the HtSH proteina in the presence of various O2 concentrations.
O2 / H2 / N2 fractionsb (% v/v) | [O2] (μM) | Hydrogenase activity in the presence of O2 (U mg−1 of protein)c | kcat (s−1) | Hydrogenase activity (%) |
---|---|---|---|---|
0 / 33.33 / 66.66 | 0.00 | 16 ± 2 | 45.9 | 100 |
0.2 / 33.33 / 66.46 | 1,9 | 15 ± 4 | 43.0 | 94.2 |
2 / 33.33 / 64.66 | 18,8 | 7.7 ± 0.3 | 21.5 | 49.8 |
10 / 33.33 / 56.66 | 94,0 | 1.3 ± 0.5 | 3.6 | 16.6 |
HtSH was purified by affinity chromatography as described in materials and methods.
For each O2 concentration, a fixed volume of H2-saturated buffer was mixed with various proportions of O2- and N2-saturated buffers. The gas phase contained the corresponding gas mixtures.
H2-mediated NAD+ reduction activity was measured at 50 °C and pH 6.5.