Literature summary extracted from
Chandrayan, S.K.; Wu, C.H.; McTernan, P.M.; Adams, M.W.
High yield purification of a tagged cytoplasmic [NiFe]-hydrogenase and a catalytically-active nickel-free intermediate form (2015), Protein Expr. Purif., 107, 90-94.
Application
EC Number |
Application |
Comment |
Organism |
---|
1.12.1.3 |
synthesis |
biohydrogen production from sugars using a mixture of enzymes in an in vitro cell-free synthetic pathway. Development of this process at an industrial scale is limited by the availability of the H2-producing enzyme |
Pyrococcus furiosus |
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
1.12.1.3 |
the four gene operon encoding the enzyme is overexpressed in Pyrococcus furiosus and includes a polyhistidine affinity tag |
Pyrococcus furiosus |
Localization
EC Number |
Localization |
Comment |
Organism |
GeneOntology No. |
Textmining |
---|
1.12.1.3 |
cytoplasm |
- |
Pyrococcus furiosus |
5737 |
- |
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
1.12.1.3 |
Pyrococcus furiosus |
Q8U2E5 and Q8U2E4 and E7FHU4 and E7FI44 |
Q8U2E5: gene PF0891, Q8U2E4: gene PF0892, E7FHU4: gene PF0893, E7FI44: gene PF0894 |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
1.12.1.3 |
one-step purification of the recombinant enzyme including a polyhistidine affinity tag results in a 50fold increase in yield compared to the four-step purification procedure for the native enzyme |
Pyrococcus furiosus |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
1.12.1.3 |
NADPH + H+ |
- |
Pyrococcus furiosus |
H2 + NADP+ |
- |
? |
|
Subunits
EC Number |
Subunits |
Comment |
Organism |
---|
1.12.1.3 |
heterotetramer |
a maturation pathway is proposed where, in the terminal step, the NiFe-containing catalytic subunit assembles with NADPH-oxidizing trimeric form to give the active holoenzym |
Pyrococcus furiosus |
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
1.12.1.3 |
[NiFe]-hydrogenase |
- |
Pyrococcus furiosus |
Temperature Optimum [°C]
EC Number |
Temperature Optimum [°C] |
Temperature Optimum Maximum [°C] |
Comment |
Organism |
---|
1.12.1.3 |
80 |
- |
assay at |
Pyrococcus furiosus |
pH Optimum
EC Number |
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
---|
1.12.1.3 |
8.4 |
- |
assay at |
Pyrococcus furiosus |