BRENDA - Enzyme Database show
show all sequences of 1.12.1.5

Engineering hyperthermophilic archaeon Pyrococcus furiosus to overproduce its cytoplasmic [NiFe]-hydrogenase

Chandrayan, S.K.; McTernan, P.M.; Hopkins, R.C.; Sun, J.; Jenney, F.E.; Adams, M.W.; J. Biol. Chem. 287, 3257-3264 (2012)

Data extracted from this reference:

Application
Application
Commentary
Organism
synthesis
the recombinant enzyme is efficient in vitro biohydrogen production
Pyrococcus furiosus
Localization
Localization
Commentary
Organism
GeneOntology No.
Textmining
cytoplasm
-
Pyrococcus furiosus
5737
-
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Iron
contains 1 nickel-iron catalytic site and 6 iron-sulfur clusters, contains 23 iron atoms/heterotetramer
Pyrococcus furiosus
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Pyrococcus furiosus
-
-
-
Pyrococcus furiosus COM1
-
-
-
Purification (Commentary)
Commentary
Organism
Strep-tag II column chromatography
Pyrococcus furiosus
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.23
-
crude native enzyme, using methyl viologen as cosubstrate, at pH 8.4 and 80°C
Pyrococcus furiosus
7.96
-
crude recombinant enzyme, using methyl viologen as cosubstrate, at pH 8.4 and 80°C
Pyrococcus furiosus
126
-
recombinant enzyme after 24fold purification, using methyl viologen as cosubstrate, at pH 8.4 and 80°C
Pyrococcus furiosus
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
H+ + NADPH
-
722777
Pyrococcus furiosus
H2 + NADP+
-
-
-
r
H+ + NADPH
-
722777
Pyrococcus furiosus COM1
H2 + NADP+
-
-
-
r
H+ + reduced methyl viologen
-
722777
Pyrococcus furiosus
H2 + oxidized methyl viologen
-
-
-
?
H+ + reduced methyl viologen
-
722777
Pyrococcus furiosus COM1
H2 + oxidized methyl viologen
-
-
-
?
Subunits
Subunits
Commentary
Organism
heterotetramer
-
Pyrococcus furiosus
Temperature Stability [°C]
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
25
90
the native enzyme shows half-lives of 14 h at 90°C under argon (H2 evolution), 21 h at 25°C under air (H2 evolution), and 10 h at 90°C under argon (H2 oxidation). The recombinant enzyme shows half-lives of 6 h at 90°C under argon (H2 evolution), 25 h at 25°C under air (H2 evolution), and 5 h at 90°C under argon (H2 oxidation)
Pyrococcus furiosus
Cofactor
Cofactor
Commentary
Organism
Structure
flavin
-
Pyrococcus furiosus
NADPH
dependent on
Pyrococcus furiosus
[2Fe-2S]-center
contains 1 [2Fe-2S] centre
Pyrococcus furiosus
[4Fe-4S]-center
contains 2 [4Fe-4S] centres
Pyrococcus furiosus
Application (protein specific)
Application
Commentary
Organism
synthesis
the recombinant enzyme is efficient in vitro biohydrogen production
Pyrococcus furiosus
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
flavin
-
Pyrococcus furiosus
NADPH
dependent on
Pyrococcus furiosus
[2Fe-2S]-center
contains 1 [2Fe-2S] centre
Pyrococcus furiosus
[4Fe-4S]-center
contains 2 [4Fe-4S] centres
Pyrococcus furiosus
Localization (protein specific)
Localization
Commentary
Organism
GeneOntology No.
Textmining
cytoplasm
-
Pyrococcus furiosus
5737
-
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Iron
contains 1 nickel-iron catalytic site and 6 iron-sulfur clusters, contains 23 iron atoms/heterotetramer
Pyrococcus furiosus
Purification (Commentary) (protein specific)
Commentary
Organism
Strep-tag II column chromatography
Pyrococcus furiosus
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.23
-
crude native enzyme, using methyl viologen as cosubstrate, at pH 8.4 and 80°C
Pyrococcus furiosus
7.96
-
crude recombinant enzyme, using methyl viologen as cosubstrate, at pH 8.4 and 80°C
Pyrococcus furiosus
126
-
recombinant enzyme after 24fold purification, using methyl viologen as cosubstrate, at pH 8.4 and 80°C
Pyrococcus furiosus
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
H+ + NADPH
-
722777
Pyrococcus furiosus
H2 + NADP+
-
-
-
r
H+ + NADPH
-
722777
Pyrococcus furiosus COM1
H2 + NADP+
-
-
-
r
H+ + reduced methyl viologen
-
722777
Pyrococcus furiosus
H2 + oxidized methyl viologen
-
-
-
?
H+ + reduced methyl viologen
-
722777
Pyrococcus furiosus COM1
H2 + oxidized methyl viologen
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
heterotetramer
-
Pyrococcus furiosus
Temperature Stability [°C] (protein specific)
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
25
90
the native enzyme shows half-lives of 14 h at 90°C under argon (H2 evolution), 21 h at 25°C under air (H2 evolution), and 10 h at 90°C under argon (H2 oxidation). The recombinant enzyme shows half-lives of 6 h at 90°C under argon (H2 evolution), 25 h at 25°C under air (H2 evolution), and 5 h at 90°C under argon (H2 oxidation)
Pyrococcus furiosus
Other publictions for EC 1.12.1.5
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
722777
Chandrayan
Engineering hyperthermophilic ...
Pyrococcus furiosus, Pyrococcus furiosus COM1
J. Biol. Chem.
287
3257-3264
2012
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1
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1
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439641
Ma
Characterization of hydrogenas ...
Pyrococcus furiosus
J. Bacteriol.
182
1864-1871
2000
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1
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5
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5
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1
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4
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4
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1
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5
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1
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1
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