Description
The [NiFe] hydrogenases contain a minimum of two subunits known as the small (S) and large (L) subunits. The small subunit contains three iron-sulfur clusters while the large subunit contains the active site, a nickel-iron center which is connected to the solvent by a molecular tunnel. To date, periplasmic, cytoplasmic, and membrane-bound hydrogenases have been found. [NiFe] hydrogenases are known to be deactivated by molecular oxygen (O2). The [NiFe] hydrogenase of Pyrococcus furiosus is heterotetrameric wherein the additional two subunits allow the enzyme to use NAD(P)(H) as an electron carrier.
Abbr
SHI, Recombinant (P. furiosus)
Source
Pyrococcus furiosus
Molecular Weight
Predicted: 155 kDa, Size Exclusion: 149 kDa +/- 5 kDa
Unit Definition
One unit (U) is 1 µmole of H2 evolved min-1 mg-1.
Thermal stability
ambient to 100°C
Storage
This item is oxygen sensitive. Stable when stored sealed in strictly anaerobic environment (<10 ppm O2) at room temperature for up to 6 months. For long-term storage, protein can be flash frozen in nitrogen and stored at -80°C.
Buffer
50 mM Tris, 2 mM DT, 300 mM NaCl, pH 8.2
Synonyms
Cytoplasmic [NiFe]-Hydrogenase; OE-SHI; Cytoplasmic Hydrogenase; NiFe-type cytoplasmic hydrogenase; SHI; [NiFe] hydrogenase
Formulation
1 mg/ml solution in 50 mM Tris-HCl, 100 mM NaCl, 5 mM DTT and 20% glycerol pH 8.0
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