Information on EC 1.2.1.43 - formate dehydrogenase (NADP+)

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

EC NUMBER
COMMENTARY hide
1.2.1.43
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RECOMMENDED NAME
GeneOntology No.
formate dehydrogenase (NADP+)
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
formate + NADP+ = CO2 + NADPH
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
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redox reaction
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reduction
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
carbon tetrachloride degradation II
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reductive acetyl coenzyme A pathway I (homoacetogenic bacteria)
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reductive acetyl coenzyme A pathway
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Methane metabolism
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Carbon fixation pathways in prokaryotes
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Metabolic pathways
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Microbial metabolism in diverse environments
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SYSTEMATIC NAME
IUBMB Comments
formate:NADP+ oxidoreductase
A tungsten-selenium-iron protein.
CAS REGISTRY NUMBER
COMMENTARY hide
51377-43-6
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
CO2 + NADPH
formate + NADP+
show the reaction diagram
formate + NAD+
CO2 + NADH
show the reaction diagram
formate + NAD+
CO2 + NADH + H+
show the reaction diagram
formate + NADP+
CO2 + NADPH
show the reaction diagram
formate + NADP+
CO2 + NADPH + H+
show the reaction diagram
additional information
?
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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
CO2 + NADPH
formate + NADP+
show the reaction diagram
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first enzyme in pathway of reduction of CO2 to acetate: CO2 serves as electron acceptor generated during fermentation
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formate + NADP+
CO2 + NADPH
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NAD+
the enzyme shows double cofactor specificity, with NADP+ preferred over NAD+ at acidic pH values
NADP+
NADPH
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Molybdenum
selenium
Tungsten
additional information
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activity of enzyme in growing cells is enhanced when selenite and molybdate are added together to the growth medium, tungstate replaces and is better than molybdate, 75Se-selenite is incorporated into protein fraction
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,3-Butanedione
5,5'-dithio-bis(2-nitrobenzoic acid)
complete inhibition at 1 mM
AgNO3
complete inhibition at 1 mM
azide
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competitive inhibitor
CdCl2
slight inhibition at 1 mM
CoCl2
slight inhibition at 1 mM
CuSO4
slight inhibition at 1 mM
cyanide
DMSO
1-33% residual activity at 40% (v/v)
ethanol
complete inhibition at 40% (v/v)
ethyl 4-chloroacetoacetate
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FeCl2
complete inhibition at 1 mM
FeSO4
formaldehyde
79.8% residual activity at 0.1 M
HgCl2
complete inhibition at 1 mM
hydroxylamine
slight inhibition at 10 mM
Hypophosphite
Isopropanol
33-66% residual activity at 20 and 40% (v/v)
mercaptoethanol
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methanol
complete inhibition at 40% (v/v)
NADP+
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substrate inhibition at 5 mM or higher concentration
NaNO3
slight inhibition at 1 mM
Sodium azide
Sodium nitrate
44.3% residual activity at 0.1 M
Sodium selenite
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0.01 M
sulfite
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inhibition of activity with NADP+ but not with methyl viologen
tert-butanol
33-66% residual activity at 10, 20 and 40% (v/v)
Triton X-100
33-66% residual activity at 5 and 10% (v/v)
additional information
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
sulfhydryl compounds
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.083 - 200
formate
2.35
methyl viologen
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0.06 - 6.5
NAD+
0.084 - 0.92
NADP+
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.25 - 5.77
NAD+
0.13 - 7.89
NADP+
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.16 - 20
NAD+
0.16 - 30
NADP+
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1.4
crude enzyme, in 1 M potassium phosphate buffer (pH 7.0), at 30°C
5.3
after 3.8fold purification, in 1 M potassium phosphate buffer (pH 7.0), at 30°C
113
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methyl viologen as electron acceptor
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6
with NADP+ as cofactor
7 - 9.5
7
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broad, CO2 reduction
7.5
with NADP as cofactor
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
70
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formate + NADP+
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40
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low activity below
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
42000
8 * 42000, SDS-PAGE
76000
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alpha2 beta2, 2 * 96000 + 2 * 76000 SDS-PAGE
79000
gel filtration
96000
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alpha2 beta2, 2 * 96000 + 2 * 76000 SDS-PAGE
270000 - 300000
340000
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
homooctamer
tetramer
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alpha2 beta2, 2 * 96000 + 2 * 76000 SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
apoprotein and holoprotein, either in complex with NAD+ or NADP+, hanging drop vapor diffusion method, using 0.1 M MES pH 6.5, 12% (w/v) PEG 2000 MME or 0. 1 M Na acetate pH 4.6, 2.0 M Na formate
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 12
more than 94% of the initial activity is retained at pH 5.0-12.0 after incubation for 30 min at 30°C
711869
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
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mutant FDH, 1 year, no loss of activity
25 - 50
the enzyme remains stable after incubation for 25 h at 25°C and 37°C, while its activity is reduced to about 35% after 3 min at 50°C and is completely lost after 15 min at 50°C
25
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mutant FDH, 7 days, no decrease of activity
55 - 60
no loss of activity is observed at 55°C for 10 h, but about half of the enzyme activity is lost after incubation for 36 h. The enzyme can maintain a 50% activity for at least 5 h when it is incubated at 60°C
73
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10 min, 60% loss of activity
76
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10 min, 90% loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
azide stabilizes during purification
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dithionite stabilizes during purification
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glycerol stabilizes during purification
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OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
extremely oxygen-sensitive
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288219, 288220, 288221, 288223, 288224
rapid inactivation by air, thiol-iron complexes and formate stabilize
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288217, 288223
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, anaerobic conditions, glycerol, ammonium sulfate, 45 days, 35% loss of activity
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DEAE-Toyopearl column chromatography and butyl-Toyopearl 650M column chromatography
HisTrap column chromatography
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large scale production in Escherichia coli; recombinant protein, expressed in Escherichia coli
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Ni-NTA column chromatography, and Superdex 200 gel filtration
recombinant protein, expressed in Escherichia coli
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Anabaena sp. PCC 7120
expressed in Escherichia coli BL21(DE3) cells
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A198G/D221Q
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the mutant has the highest catalytic efficiency with NADP+
C145S/A198G/D221Q/C255V
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the mutant has high catalytic efficiency with NADP+
C145S/D221A/C255V
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the mutant has high catalytic efficiency with NADP+
C145S/D221Q/C255V
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the mutant shows the highest specific activity for a NADP+-accepting formate dehydrogenase
D221A
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the mutant has high catalytic efficiency with NADP+
D221G
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the mutant has high catalytic efficiency with NADP+
D221Q
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the mutant has high catalytic efficiency with NADP+
D221S
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the mutant has high catalytic efficiency with NADP+
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis