Information on EC 1.12.1.2 - hydrogen dehydrogenase

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The expected taxonomic range for this enzyme is: Bacteria, Archaea

EC NUMBER
COMMENTARY hide
1.12.1.2
-
RECOMMENDED NAME
GeneOntology No.
hydrogen dehydrogenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
H2 + NAD+ = H+ + NADH
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
hydrogen oxidation II (aerobic, NAD)
-
-
hydrogen production II
-
-
hydrogen production
-
-
SYSTEMATIC NAME
IUBMB Comments
hydrogen:NAD+ oxidoreductase
An iron-sulfur flavoprotein (FMN or FAD). Some forms of this enzyme contain nickel.
CAS REGISTRY NUMBER
COMMENTARY hide
37256-54-5
deleted
9027-05-8
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain 4a-2
-
-
Manually annotated by BRENDA team
strain 4a-2
-
-
Manually annotated by BRENDA team
strain B19
-
-
Manually annotated by BRENDA team
strain Cd2/01
-
-
Manually annotated by BRENDA team
strain G27
-
-
Manually annotated by BRENDA team
strain N9A
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
isolates of nine different actinorhizal host plants, 18 strains, overview
-
-
Manually annotated by BRENDA team
strain CALU 743
-
-
Manually annotated by BRENDA team
strain CALU 743
-
-
Manually annotated by BRENDA team
Hydrogenomonas sp.
strain H16
-
-
Manually annotated by BRENDA team
strain H16
-
-
Manually annotated by BRENDA team
methanotroph, constitutive soluble hydrogenase
-
-
Manually annotated by BRENDA team
no activity in Alcaligenes paradoxus
-
-
-
Manually annotated by BRENDA team
no activity in Aquaspirillum autotrophicum
-
-
-
Manually annotated by BRENDA team
no activity in Corynebacterium autotrophicum
-
-
-
Manually annotated by BRENDA team
no activity in Hydrogenophaga palleronii
-
-
-
Manually annotated by BRENDA team
no activity in Paracoccus denitrificans
-
-
-
Manually annotated by BRENDA team
no activity in Pseudomonas facilis
-
-
-
Manually annotated by BRENDA team
strain MR11
-
-
Manually annotated by BRENDA team
synthetic construct
-
-
-
Manually annotated by BRENDA team
strain KOD1
-
-
Manually annotated by BRENDA team
strain BBS
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2 ferricyanide + NAD(P)H
2 ferrocyanide + NAD(P)+ + H+
show the reaction diagram
ferrocyanide + NAD(P)H
ferricyanide + NAD(P)+
show the reaction diagram
-
enzyme complex including HoxI
-
-
?
ferrocyanide + NAD+
ferricyanide + NADH
show the reaction diagram
H+ + NADH
H2 + NAD+
show the reaction diagram
H+ + NADH + reduced ferredoxin
H2 + NAD+ + oxidized ferredoxin
show the reaction diagram
H+ + NADPH
H2 + NADP+
show the reaction diagram
-
-
-
-
?
H+ + reduced dithionite
H2 + oxidized dithionite
show the reaction diagram
H+ + reduced methyl viologen
H2 + oxidized methyl viologen
show the reaction diagram
H2
H+ + e-
show the reaction diagram
H2 + acceptor
H+ + reduced acceptor
show the reaction diagram
H2 + ferricyanide
H+ + ferrocyanide
show the reaction diagram
H2 + ferrocyanide
H+ + ferricyanide
show the reaction diagram
-
-
-
-
r
H2 + NAD(P)+
H+ + NAD(P)H
show the reaction diagram
-
-
-
r
H2 + NAD+
H+ + NADH
show the reaction diagram
H2 + NADP+
H+ + NADPH
show the reaction diagram
H2 + oxidized benzyl viologen
H+ + reduced benzyl viologen
show the reaction diagram
H2 + oxidized benzyl viologen
reduced benzyl viologen + H+
show the reaction diagram
H2 + oxidized dithionite
H+ + reduced dithionite
show the reaction diagram
-
-
-
r
H2 + oxidized methyl viologen
H+ + reduced methyl viologen
show the reaction diagram
H2 + oxidized methyl viologen
reduced methyl viologen + H+
show the reaction diagram
H2 + oxidized methylene blue
H+ + reduced methylene blue
show the reaction diagram
NAD(P)H + H+ + oxidized 2,6-dichlorophenolindophenol
NAD(P)+ + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
NAD+ + H+ + e-
NADH
show the reaction diagram
-
diaphorase reaction part
-
-
r
NADH + K3Fe(CN)6
?
show the reaction diagram
-
-
-
-
?
NADH + oxidized 2,6-dichlorophenol indophenol
NAD+ + reduced 2,6-dichlorophenol indophenol
show the reaction diagram
NADH + oxidized benzyl viologen
NAD+ + reduced benzyl viologen
show the reaction diagram
-
-
-
-
r
NADH + reduced 3-acetylpyridine adenine dinucleotide
NAD+ + oxidized 3-acetylpyridine adenine dinucleotide
show the reaction diagram
NADPH + K3Fe(CN)6
?
show the reaction diagram
-
low reaction with hexameric enzyme form, no reaction with tetrameric enzyme form
-
-
?
oxidized benzyl viologen + NADH
reduced benzyl viologen + NAD+
show the reaction diagram
oxidized cytochrome c + NAD(P)H
reduced cytchrome c + NAD(P)+
show the reaction diagram
-
-
-
?
oxidized methylene blue + NAD(P)H
reduced methylene blue + NAD(P)+
show the reaction diagram
-
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
H+ + NADH
H2 + NAD+
show the reaction diagram
-
-
-
-
?
H+ + NADPH
H2 + NADP+
show the reaction diagram
-
-
-
-
?
H2 + NAD+
H+ + NADH
show the reaction diagram
H2 + oxidized methyl viologen
H+ + reduced methyl viologen
show the reaction diagram
P22317, P22318, P22319, P22320
the artificial electron donor is accepted by the cells for H2 production when added to the culture medium
-
-
r
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4Fe-4S-center
presence of a 4Fe-4S-center in addition to the active site iron; presence of a 4Fe-4S-center in addition to the active site iron
ATP
-
30% stimulation
benzyl viologen
-
oxidized and reduced
Ferredoxin
-
the hydrogenase requires the presence of both electron carriers, NADH and reduced ferredoxin, synergistically for catalysis of H2 production
-
flavin
iron-sulfur centre
NAD(P)H
NADH is 2times more active than NADPH
[2Fe-2S]-center
HoxFU accommodates a [2Fe2S] cluster, FMN and a series of [4Fe4S] clusters; HoxFU accommodates a [2Fe2S] cluster, FMN and a series of [4Fe4S] clusters
[4Fe-4S]-center
HoxFU accommodates a [2Fe2S] cluster, FMN and a series of [4Fe4S] clusters; HoxFU accommodates a [2Fe2S] cluster, FMN and a series of [4Fe4S] clusters
additional information
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
-
NAD+ reduction with H2 is completely dependent on the presence of divalent metal ions Ni2+, Co2+, Mg2+ or Mn2+ or of high salt concentrations between 500-1500 mM
cyanide
-
enzyme contains four cyanides in its active site, one of which is responsible for the insensitivity towards oxygen
K+
-
activates
Nickel
-
[NiFe] hydrogenases carry a metal centre composed of Fe and Ni atoms at the active site
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
CdSO4
-
1 mM, complete inhibition
CuCl2
-
0.2 mM, complete inhibition
cyanide
-
1 mM, 33% inhibition
diethyl dicarbonate
-
chemical modification of active site His residues results in reduced enzymatic hydrogenase and diaphorase activities, pH-dependence and kinetics of modifications/inactivation
DTNB
-
modification of thiol groups of activated enzyme results in rapid inactivation of both activities, modification of the residues of nonactivated enzyme has small effects on the enzyme activities, pH-dependence and kinetics of modifications/inactivation
glutathione
-
above 0.1 mM: inactivation, up to 0.1 mM: activation
HgCl2
-
0.001 mM, complete inhibition
High ionic strength
-
200 mM phosphate buffer, 70% inhibition
-
iodoacetate
iodoacetic acid
-
modification of thiol groups of activated enzyme results in rapid inactivation of both activities, modification of the residues of nonactivated enzyme has small effects on the enzyme activities, pH-dependence and kinetics of modifications/inactivation
mercaptoethanol
Hydrogenomonas sp.
-
-
NAD+
product inhibition; product inhibition
NADH
product inhibition; product inhibition
Ni2+
-
0.5-1 mM, strong inhibition of artificial electron acceptor reduction
p-chloromercuribenzoate
Sulfide
Triethanolamine buffer
-
at high concentration
-
Tris-buffer
-
at high concentration
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
glutathione
-
up to 0.1 mM: activation, above 0.1 mM: inactivation
HypC
-
[NiFe] hydrogenase maturation protein HypC catalyzes the insertion and cyanation of the iron center of [NiFe] hydogenase
-
HypD
-
[NiFe] hydrogenase maturation protein HypD catalyzes the insertion and cyanation of the iron center of [NiFe] hydogenase
-
HypE
-
[NiFe] hydrogenase maturation protein HypE catalyzes the insertion and cyanation of the iron center of [NiFe] hydogenase
-
LexA protein
-
the transcription factor LexA contributes to enzyme expression
-
Na2S2O4
-
1 mM, activity of initially inactive enzyme increases within 1 h to a stable level in the presence of 101 kPa H2
NADPH
-
activates hexameric enzyme form. Subunit HoxI provides a binding domain for NADPH; specific activation of the soluble oligomeric enzyme, the binding domain is located on subunit HoxI
NH4+
-
activates
phosphate
-
increase in NAD-reduction rate
Sll0359 protein
-
the AbrB-like protein Sll0359 interacts specifically with the hox promoter region and works as an activator of hox gene expression
-
SO42-
-
activates
[NiFe] hydrogenase maturation protein
-
maturation of the [NiFe] hydrogenase involves assembly of nonproteinaceous ligands on the large subunit by accessory proteins encoded by the hyp operon, HypE is an essential accessory protein and participates in the synthesis of two cyano groups found in the large subunit
-
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.44
2,6-dichlorophenolindophenol
-
-
4.1
benzyl viologen
-
-
0.38
FAD
-
-
0.45 - 4
ferricyanide
0.037 - 0.19
H2
0.05 - 5.9
methyl viologen
0.25 - 0.44
methylene blue
0.0035 - 0.56
NAD+
0.0023 - 0.07
NADH
0.0007
NADPH
-
30C, pH 7.0
0.0578
Oxidized benzyl viologen
-
30C, pH 8.5
0.0375
oxidized methyl viologen
-
30C, pH 8.5
0.00025
reduced ferredoxin
-
pH and temperature not specified in the publication
0.0306 - 0.089
reduced methyl viologen
0.028
resazurin
-
-
additional information
additional information
-
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2 - 372
ferrocyanide
70 - 372
H2
109 - 143
NAD+
171 - 222
NADH
2
NADPH
Cupriavidus necator
-
pH 8.0, wild-type enzyme complex including HoxI, with ferrocyanide
70 - 120
Oxidized benzyl viologen
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.2
NAD+
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0006
0.0017
-
soluble periplasmic fraction, hydrogen producing activity with electron acceptor oxidized methyl viologen
0.0018
cell culture in nitrogen-limited medium, anaerobic conditions, H2 production using reduced methyl viologen as electron donor
0.0029
-
soluble periplasmic fraction, hydrogen producing activity with electron donor NADH
0.006
-
soluble periplasmic fraction, hydrogen producing activity with electron donor reduced benzyl viologen
0.0095
-
soluble periplasmic fraction, hydrogen producing activity with electron acceptor oxidized benzyl viologen
0.0113
-
soluble periplasmic fraction, hydrogen producing activity with electron donor reduced methyl viologen
0.0175
-
soluble periplasmic fraction, hydrogen producing activity with electron acceptor oxidized methylene blue
0.0255
-
soluble periplasmic fraction, hydrogen producing activity with electron acceptor NAD+
0.035
-
H2 producing activity, strain R43 grown without nitrogen and nickel, from host plant Compotonia cunninghamiana, with reduced methyl viologen as electron donor
0.076
-
crude extract, pH and temperature not specified in the publication
0.11
-
H2 producing activity, strain UGL011101 grown without nitrogen and nickel, from host plant Alnus incana, with reduced methyl viologen as electron donor
0.3
-
H2 producing activity, strain HFPCcI3 grown without nitrogen and nickel, from host plant Compotonia cunninghamiana, with reduced methyl viologen as electron donor
1.7
-
recombinant enzyme expressed in Ralstonia eutropha
4.91
-
H2 producing activity, strain UGL140102 grown without nitrogen and nickel, from host plant Hippophae rhamnoides, with reduced methyl viologen as electron donor
5
-
purified enzyme, H2 consumption
5.6
-
strain Cd2/01, electron acceptor NAD+
7.7
-
electron acceptor NAD+
10
-
purified enzyme, H2 production
15
native purified enzyme complex from strain PCC 6301, hydrogen-forming activity, cofactor reduced methyl viologen
17 - 84
-
activity of different batches with NAD+ and H2
25.2
-
reaction with H2 and benzyl viologen, purified HoxI deletion mutant enzyme
30 - 100
-
purified enzyme, forward reaction under aerobic conditions with NAD+
34
-
electron acceptor NAD+
35.1
-
reaction with H2 and benzyl viologen, purified wild-type enzyme complex including HoxI
36.5
Hydrogenomonas sp.
-
-
38
-
strain H16, electron acceptor NAD+
39
-
reaction with H2 and NAD+, purified HoxI deletion mutant enzyme
41.9
-
reaction with H2 and NAD+, purified wild-type enzyme complex including HoxI
61.6
-
reaction with NADH and ferrocyanide, purified HoxI deletion mutant enzyme
64.9
-
reaction with NADH and ferrocyanide, purified wild-type enzyme complex including HoxI
67
native purified enzyme complex from strain PCC 6803, hydrogen-forming activity, cofactor reduced methyl viologen
72
-
electron acceptor NAD+
85
-
electron acceptor NAD+
87.78
-
after 1155old purification, pH and temperature not specified in the publication
100
-
electron acceptor benzyl viologen
102.1
-
reaction with H2 and ferrocyanide, purified HoxI deletion mutant enzyme
108.8
-
reaction with H2 and ferrocyanide, purified wild-type enzyme complex including HoxI
125 - 175
-
activity of different batches with NADH and ferricyanide
230
-
recombinant protein, pH 8.0, temperature not specified in the publication
1263
-
activity of different batches with benzyl viologen and H2
1300
-
purified enzyme, with substrates oxidized benzyl viologen and NADH
1700
-
purified enzyme, both reaction directions with substrates oxidized/reduced methyl viologen and H2/H+
2030
-
purified hydrogenase, at 30C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.3
-
evolution of H2 in the presence of reduced methyl viologen
6
-
electron donor NADH
6.1
-
electron acceptor NAD+, no addition of Ni2+, Tris/MES buffer
6.3
strain PCC 6803, hydrogen-forming activity
6.8
-
evolution of H2 in the presence of NADH or NADPH
7.8 - 8
-
electron acceptor NAD+, in triethanolamine/HCl buffer containing NiCl2
8 - 8.2
-
electron acceptor NAD+
9
-
evolution of H2 in the presence of methyl viologen or benzyl viologen
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.9 - 9.1
Hydrogenomonas sp.
-
Tris-HCl buffer and potassium phosphate buffer pH 6.9: 20% activity, Tris-HCl buffer pH 9.1: 40% activity, potassium phosphate buffer pH 9.1: 65% activity
7 - 9.5
-
triethanolamine buffer, Tris buffer pH 7.0: 40-45% activity, triethanolamine buffer, Tris buffer pH 9.5: 60-75% activity
additional information
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20
-
assay at
30
-
assay at
36
Hydrogenomonas sp.
-
-
45
-
evolution of H2 in the presence of NADH or NADPH
50
-
evolution of H2 in the presence of NADH or NADPH
60
strain PCC 6803, hydrogen-forming activity
65
-
evolution of H2 in the presence of reduced methyl viologen
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
162500
-
amino acid sequence calculation
178000
-
sucrose density gradient centrifugation
200000
-
gel filtration
204500
-
gel filtration
206000
-
sucrose density gradient centrifugation
375000
native dimeric assembly of the pentameric enzyme complex, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexamer
oligomer
tetramer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
six accessory proteins HypA-F (hydrogenase pleiotropic proteins) and one protease (HoxW) are necessary for post-translational processing of the bidirectional [NiFe] hydrogenase
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapour diffusion method, using sodium citrate and imidazole
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6
-
1 day: 9% loss of activity in the presence of 0.5 mM NiCl2 + 5 mM MgCl2, 56% loss of activity in the absence of NiCl2 and MgCl2, 3 days: 44% loss of activity in the presence of 0.5 mM NiCl2 + 5 mM MgCl2, 80% loss of activity in the absence of NiCl2 and MgCl2
439702
6.2 - 7
-
stability of the enzyme complex including HoxI in potassium phosphate buffer at pH 6.2 and pH 7.0, not at pH 8.0 or in Tris-HCl buffer at pH 7.2 and pH 8.0, or in Tris-nitrate buffer at pH 7.4
655890
6.5
-
highest stability, 4C, H2-atmosphere, 3 days
439701
7
-
1 day: no loss of activity in the presence of 0.5 mM NiCl2 + 5 mM MgCl2, 62% loss of activity in the absence of NiCl2 and MgCl2, 3 days: 21% loss of activity in the presence of 0.5 mM NiCl2 + 5 mM MgCl2, 87% loss of activity in the absence of NiCl2 and MgCl2
439702
8
-
1 day: no loss of activity in the presence of 0.5 mM NiCl2 + 5 mM MgCl2, 86% loss of activity in the absence of NiCl2 and MgCl2, 3 days: 27% loss of activity in the presence of 0.5 mM NiCl2 + 5 mM MgCl2, complete loss of activity in the absence of NiCl2 and MgCl2
439702
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35
-
pH 8.0, native enzyme, half-life 5.3 h
45
-
enzyme prepared in Tris or triethanolamine buffer: 90% loss of activity after 10 min, enzyme prepared in K-phosphate buffer: 30% loss of activity after 10 min
50
-
thermoinactivation by dissociation of the native enzyme tetramer into constituent heterodimers
75
inactivation above
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
5 mM Mg2+ and 0.5 mM Ni2+ stabilizes
-
addition of 5 mM Mg2+ + 0.5 mM Ni2+ stabilizes
-
highly diluted enzyme preparations are significantly inactivated even at 20C
-
Ni2+ stabilizes
-
oxidized inactive form of the enzyme is the most stable against thermodenaturation, proteolysis, and inactivation in urea
-
unstable in reduced active form
-
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
air the presence of NADH leads to rapid destruction of the hexameric enzyme
-
674924
enzyme is oxygen-labile
654821
highly insensitive to O2
-
673177
insensitive to O2
-
656298
insensitivity towards oxygen, irreversible enzyme inhibition by oxygen occurs if the CN- bound to Ni2+ is irreversibly removed or if the enzyme is reduced by NADH
-
656298
oxidation with O2 and ferricyanide in absence of reducing components stabilizes, simultaneous presence of H2, NADH and O2 causes irreversible inactivation, half-lives in H2/O2 mixtures: 5% O2, 60 min, 20% O2, 30 min, 50% O2, 12 min
-
439708
oxidized hydrogenase purified under aerobic conditions is highly stable but not reactive, reductive activation of the enzyme by H2 in the presence of catalytic amounts of NADH or by reducing agents destabilizes
-
439699
rapid and irreversible inactivation under aerobic conditiones in the presence of reductants e.g. 1-10 mM NADH
-
439704
redox-dependent inactivation and activation, proposed inactivation mechanism
-
439704, 439705
the hexameric SH preparation is both active and stable in the presence of NADPH and air
-
674924
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-16C, 10 d, 30% loss of activity
Hydrogenomonas sp.
-
-20C, 4 mg/ml protein concentration, pH 7.0
-20C, O2, several months, no loss in activity
-
4C, 500 mM potassium phosphate, pH 7.0, 3 d, 10% loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
20fold by several chromatographic steps
-
hexameric enzyme form; recombinant Strep-tagged HoxI-deletion mutant tetramer from Escherichia coli by 2 different procedures
-
improved purification procedure
-
partially, preparation of periplasmic fraction
-
protamine sulfate, ammonium sulfate, DEAE-cellulose, Sephadex G-200
-
recombinant protein. The as-isolated module HoxHY is initially catalytically inactive, but after reductive activation at low potentials, exhibits both H2 oxidation and H+ reduction; recombinant protein. The as-isolated module HoxHY is initially catalytically inactive, but after reductive activation at low potentials, exhibits both H2 oxidation and H+ reduction
sp. H16, cetavlon, ammonium sulfate, DEAE-cellulose, Sephadex G-200, hydroxyapatite
-
Strep-Tactin-Sepharose column chromatography
-
under anaerobic conditions, pentameric bidirectional hydrogenase complex HoxEFUYH to near homogeneity 647fold from strain PCC 6803 and 1290fold from strain PCC 6301, recombinant His-tagged HoxE from Escherichia coli
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, HoxE is encoded in a cluster of 5 genes encoding the enzyme complex subunits, sigma54-type promotor with probably inducible expression of the operon
expression in Cupriavidus necator; expression in Cupriavidus necator
gene cluster HoxFUYHI, cloning and expression of the Strep-tagged HoxI-deletion mutant, a tetramer composed of HoxFU and HoxHY dimers forming a tetramer in Escherichia coli strains JM109 and XL-1 Blue
-
heterologous expression in Ralstonia eutropha
-
over-expression of a plasmid carrying the hoxFUYWI-hypA2B2F2CDEXA genes in Cupriavidus necator
-
overexpression of HoxE in Escherichia coli strain M15
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C461A
-
site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, inactive
D456N
-
site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, nearly inactive
D456S
-
site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, nearly inactive
E14Q
-
site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, inactive
E14V
-
site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, nearly inactive
G15A
-
site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, nearly inactive
H396L
-
site-directed mutagenesis, catalytic subunit HoxH allele, highly reduced activity compared to the wild-type strain
H396Q
-
site-directed mutagenesis, catalytic subunit HoxH allele, about 3.5fold reduced activity compared to the wild-type strain
H69Q
-
site-directed mutagenesis, catalytic subunit HoxH allele, about 5fold reduced activity compared to the wild-type strain
H69Q/P390A
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site-directed mutagenesis, catalytic subunit HoxH alleles, reduced activity compared to the wild-type strain
I64A
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site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, highly reduced activity compared to the wild-type strain
L118A
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site-directed mutagenesis, catalytic subunit HoxH allele, reduced activity compared to the wild-type strain
L118F
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site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow at O2 concentrations above 5%, about 4fold reduced activity compared to the wild-type strain
L118I
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site-directed mutagenesis, catalytic subunit HoxH allele, reduced activity compared to the wild-type strain
L394A
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site-directed mutagenesis, catalytic subunit HoxH allele, about 5fold reduced activity compared to the wild-type strain
L394N
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site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, nearly inactive
P390A
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site-directed mutagenesis, catalytic subunit HoxH allele, about 5fold reduced activity compared to the wild-type strain
R12L
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site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, highly reduced activity compared to the wild-type strain
R12Q
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site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, highly reduced activity compared to the wild-type strain
R391L
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site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, inactive
R391Q
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site-directed mutagenesis, catalytic subunit HoxH allele, mutant strain does not grow, inactive
R60Q
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site-directed mutagenesis, catalytic subunit HoxH allele, highly reduced activity compared to the wild-type strain
S68V
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site-directed mutagenesis, catalytic subunit HoxH allele, highly reduced activity compared to the wild-type strain
T415S
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site-directed mutagenesis, catalytic subunit HoxH allele, about 5fold reduced activity compared to the wild-type strain
T415V
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site-directed mutagenesis, catalytic subunit HoxH allele, highly reduced activity compared to the wild-type strain
T415V/N415H
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site-directed mutagenesis, catalytic subunit HoxH alleles, O2-sensitive growth, highly reduced activity compared to the wild-type strain
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
Show AA Sequence (440 entries)
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