Recombinant Pyrococcus furiosus NADP-specific NiFe-type cytoplasmic hydrogenase (4 subunits), 9×His tagged

• Cat.No.: SHI-61P
• Product Overview: This recombinant Pyrococcus furiosus NADP-specific NiFe-type cytoplasmic hydrogenase (SHI) is from the hyperthermophilic P. furiosus microorganism that grows optimally at 100 centigrade. The enzyme is active over a broad temperature range, from ambient to 100 centigrade, and is oxygen-sensitive, with a half-life in air of approximately 25 hours at room temperature.

Specification

• Species: Pyrococcus furiosus
• Source: Pyrococcus furiosus
• Tag: 9×His
• Description: This is an extremely thermostable, but oxygen sensitive enzyme. 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.
• Bio-activity: One unit (U) is 1 μmole of H2 evolved min-1 mg-1 using MV-linked assay.
• Molecular Mass: Predicted: 155 kDa, Size Exclusion: 149 kDa +/- 5 kDa
• Purity: > 90%
• Unit Definition: 1 μmole of H2 evolved min-1 mg-1 using MV-linked assay.
• 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 centigrade.
• Concentration: 1 mg/mL (> 100 U/mL)
• Storage Buffer: 50 mM Tris, 2 mM DT, 300 mM NaCl, pH 8.2
• Shipping: Dry ice
• References: 1. Ma, K. and Adams, M. W. W. (2001) Hydrogenases I and II from Pyrococcus furiosus Meths. Enzymol. 331, 208-216
  2. Chandrayan, S. K., McTernan, P. M., Hopkins, R. C., Sun, J., Jenney, F. E., Jr. & Adams, M. W. (2012) Engineering hyperthermophilic archaeon Pyrococcus furiosus to overproduce its cytoplasmic [NiFe]-hydrogenase J. Biol. Chem. 287, 3257-64
  3. Hopkins, R.C., Sun, J., Jenney, F.E., Chandrayan, S.K., McTernan, P.M., and Adams, M.W.W., (2011) "Homologous expression of a subcomplex of Pyrococcus furiosus hydrogenase that interacts with pyruvate ferredoxin oxioreductase" PLoS ONE 6(1), e26569
  4. Patent Pending - University of Georgia research Foundation, Inc. (WO 2009075798)

Gene Information

• Official Symbol: SHI
• Synonyms: NADP-specific NiFe-type cytoplasmic hydrogenase; SHI; NADPH-dependent hydrogenase/sulfhydrogenase 1 subunit alpha; NADPH-dependent hydrogenase/sulfhydrogenase 1 subunit beta; NADPH-dependent hydrogenase/sulfhydrogenase 1 subunit gamma; NADPH-dependent hydrogenase/sulfhydrogenase 1 subunit delta; Cytoplasmic [NiFe]-Hydrogenase (4 subunits; OE-SHI)