Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli. The native Escherichia coli maturation machinery will also generate a functional hydrogenase when provided with only the genes encoding the hydrogenase subunits and a single protease from Pyrococcus furiosus. The expression is induced by anaerobic conditions, whereby Escherichia coli is grown aerobically and production of recombinant hydrogenase is achieved by simply changing the gas feed from air to an inert gas (N2). The recombinant enzyme is purified and shown to be functionally similar to the native enzyme purified from Pyrococcus furiosus. The methodology to generate this key hydrogen-producing enzyme has dramatic implications for the production of hydrogen and NADPH as vehicles for energy storage and transport, for engineering hydrogenase to optimize production and catalysis, as well as for the general production of complex, oxygen-sensitive metalloproteins | Pyrococcus furiosus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.007 | - |
NADPH | 80°C, pH 8.4, recombinant enzyme | Pyrococcus furiosus | |
0.03 | - |
NADPH | 80°C, pH 8.4, native enzyme | Pyrococcus furiosus |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytoplasm | - |
Pyrococcus furiosus | 5737 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Iron | - |
Pyrococcus furiosus | |
Nickel | - |
Pyrococcus furiosus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
150000 | - |
gel filtration | Pyrococcus furiosus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pyrococcus furiosus | E7FI44 AND Q8U2E5 AND Q8U2E4 AND E7FHU4 | E7FI44: subunit alpha, Q8U2E5: subunit beta, Q8U2E4: subunit gamma, E7FHU4: subunit delta | - |
Oxidation Stability | Organism |
---|---|
t1/2 for the native enzyme is 25 h at 23°C in presence of air, t1/2 for for the recombinant enzyme is 20 h at 23°C in presence of air | Pyrococcus furiosus |
Purification (Comment) | Organism |
---|---|
- |
Pyrococcus furiosus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
H+ + NADPH | - |
Pyrococcus furiosus | H2 + NADP+ | - |
? | |
H+ + reduced benzyl viologen | - |
Pyrococcus furiosus | H2 + oxidized benzyl viologen | - |
? |
Synonyms | Comment | Organism |
---|---|---|
NADP-dependent hydrogenase | - |
Pyrococcus furiosus |
SHI | - |
Pyrococcus furiosus |
soluble hydrogenase I | - |
Pyrococcus furiosus |
[NiFe]-hydrogenase | - |
Pyrococcus furiosus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
80 | - |
assay at | Pyrococcus furiosus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
90 | - |
t1/2: 30 h for the native enzyme, t1/2: 1 h for the recombinant enzyme | Pyrococcus furiosus |
100 | - |
t1/2: 2 h, native enzyme | Pyrococcus furiosus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.4 | - |
assay at | Pyrococcus furiosus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
[Ni-Fe]-center | metal content Ni:Fe is 1:25 for the native enzyme and 1:32 for the recombinant enzyme | Pyrococcus furiosus |