Information on EC 1.18.1.3 - ferredoxin-NAD+ reductase

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The expected taxonomic range for this enzyme is: Bacteria

EC NUMBER
COMMENTARY
1.18.1.3
-
RECOMMENDED NAME
GeneOntology No.
ferredoxin-NAD+ reductase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
2 reduced [2Fe-2S] ferredoxin + NAD+ + H+ = 2 oxidized [2Fe-2S] ferredoxin + NADH
show the reaction diagram
(1)
-
-
-
reduced 2[4Fe-4S] ferredoxin + NAD+ + H+ = oxidized 2[4Fe-4S] ferredoxin + NADH
show the reaction diagram
(2)
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Fatty acid degradation
-
Xylene degradation
-
SYSTEMATIC NAME
IUBMB Comments
ferredoxin:NAD+ oxidoreductase
Contains FAD. Reaction (1) is written for a [2Fe-2S] ferredoxin, which is characteristic of some mono- and dioxygenase systems. The alternative reaction (2) is written for a 2[4Fe-4S] ferredoxin, which transfers two electrons, and occurs in metabolism of anaerobic bacteria.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
BphA4
Acidovorax sp.
-
-
ferredoxin NADPH reductase
-
-
ferredoxin-linked NAD reductase
-
-
-
-
ferredoxin-NAD reductase
-
-
-
-
ferredoxin-NAD(P)H reductase
-
-
ferredoxin-NAD(P)H reductase
-
-
-
ferredoxin-NADH oxidoreductase
Novosphingobium sp.
-
-
ferredoxin-NADH oxidoreductase
Novosphingobium sp. KA1
-
-
-
ferredoxin-reductase
-
-
NAD-ferredocinTOL reductase
-
component of toluene dioxygenase
NAD-ferredoxin reductase
-
-
-
-
NADH flavodoxin oxidoreductase
-
-
-
-
NADH-dependent ferredoxin reductase
Acidovorax sp.
-
-
NADH-ferredoxin oxidoreductase
-
-
-
-
NADH-ferredoxin reductase
-
-
-
-
NADH-ferredoxinNAP reductase
-
component of naphthalene dioxygenase multicomponent enzyme system
NADH-ferredoxinNAP reductase
Pseudomonas sp. NCIB 9816
-
component of naphthalene dioxygenase multicomponent enzyme system
-
NADH2-ferredoxin oxidoreductase
-
-
-
-
palustrisredoxin reductase
-
-
palustrisredoxin reductase
-
-
-
Red
Novosphingobium sp.
-
-
Red
Novosphingobium sp. KA1
-
-
-
RedIIA
Novosphingobium sp.
-
-
RedIIA
Novosphingobium sp. KA1
-
-
-
reductase, ferredoxin
-
-
-
-
reductase, ferredoxin-nicotinamide adenine dinucleotide
-
-
-
-
reductase, reduced nicotinamide adenine dinucleotide-ferredoxin
-
-
-
-
ferredoxin-reductase
-
-
-
additional information
-
the enzyme is a member of the oxygenase-coupled NADH-dependent ferredoxin reductase family of FAD-dependent electron-transfer enzymes
additional information
-
the enzyme is a member of the oxygenase-coupled NADH-dependent ferredoxin reductase family of FAD-dependent electron-transfer enzymes
-
CAS REGISTRY NUMBER
COMMENTARY
39369-37-4
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
Acidovorax sp.
Pseudomonas sp., strain KKS102
-
-
Manually annotated by BRENDA team
strain CNRZ 510
-
-
Manually annotated by BRENDA team
Clostridium tyrobutyricum CNRZ
strain CNRZ 510
-
-
Manually annotated by BRENDA team
Novosphingobium sp.
carAd or fdr gene
-
-
Manually annotated by BRENDA team
Novosphingobium sp. KA1
carAd or fdr gene
-
-
Manually annotated by BRENDA team
NAD-ferredocinTOL reductase component of toluene dioxygenase
-
-
Manually annotated by BRENDA team
NADH-ferredoxin oxidoreductase component of biphenyl 2,3-dioxygenase; strain LB400
-
-
Manually annotated by BRENDA team
NADH-ferredoxinNAP reductase component of naphthalene dioxygenase; strain NCIB 9816
-
-
Manually annotated by BRENDA team
strain KKS102
-
-
Manually annotated by BRENDA team
Pseudomonas sp. LB400
strain LB400
-
-
Manually annotated by BRENDA team
Pseudomonas sp. NCIB 9816
strain NCIB 9816
-
-
Manually annotated by BRENDA team
two different isoforms
-
-
Manually annotated by BRENDA team
activity is present in wild-type strain, alcohol-adapted strain lacks detectable levels of reduced ferredoxin-linked NAD reductase
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
Novosphingobium sp.
-
the ferredoxin-NADH oxidoreductase Red is part of the carbazole 1,9a-dioxygenase. Red and is classified as a class IIA Rieske oxygenase
additional information
Novosphingobium sp. KA1
-
the ferredoxin-NADH oxidoreductase Red is part of the carbazole 1,9a-dioxygenase. Red and is classified as a class IIA Rieske oxygenase
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ferredoxin + NADH + H+
reduced ferredoxin + NAD+
show the reaction diagram
Acidovorax sp.
-
BphA3, a Rieske-type [2Fe-2S] ferredoxin, and BphA4 comprise an electron-transfer system for the multi-component dioxygenase BphA, BphA4 receives two electrons from NADH as a form of hydride and transfers an electron to each of two BphA3 molecules, molecular mechanism of the electron transfer between BphA3 and BphA4, overview
-
-
?
ferredoxin + NADH + H+
reduced ferredoxin + NAD+
show the reaction diagram
-
the enzyme is an essential component of the cytochrome P450 systems in Rhodopseudomonas palustris strain CGA009, a model organism with diverse metabolic pathways, electron transport is initiated by a single two-electron transfer from NAD(P)H to FAD to produce fully reduced FADH2, which provides one electron to the iron-sulfur cluster, in most cases a [2Fe2S] cluster, of ferredoxins. These electrons are finally transferred to the terminal oxygenase, mechanism of electron transport, overview
-
-
?
NADH + 2,6-dichlorophenolindophenol
NAD+ + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
-
-
-
-
?
NADH + 2,6-dichlorophenolindophenol
NAD+ + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
-
-
-
-
?
NADH + 2,6-dichlorophenolindophenol
NAD+ + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
Pseudomonas sp. LB400
-
-
-
-
?
NADH + 2,6-dichlorophenolindophenol
NAD+ + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
Pseudomonas sp. NCIB 9816
-
-
-
-
?
NADH + cytochrome c
NAD+ + reduced cytochrome c
show the reaction diagram
-
-
-
-
?
NADH + cytochrome c
NAD+ + reduced cytochrome c
show the reaction diagram
-
-
-
-
?
NADH + cytochrome c
NAD+ + reduced cytochrome c
show the reaction diagram
-
-
-
-
?
NADH + cytochrome c
NAD+ + reduced cytochrome c
show the reaction diagram
Pseudomonas sp. LB400
-
-
-
-
?
NADH + cytochrome c
NAD+ + reduced cytochrome c
show the reaction diagram
-
-
-
-
?
NADH + cytochrome c
NAD+ + reduced cytochrome c
show the reaction diagram
Pseudomonas sp. NCIB 9816
-
-
-
-
?
NADH + ferricyanide
NAD+ + ?
show the reaction diagram
-
-
-
-
?
NADH + ferricyanide
NAD+ + ?
show the reaction diagram
-
-
-
-
?
NADH + ferricyanide
NAD+ + ?
show the reaction diagram
-
-
-
-
?
NADH + ferricyanide
NAD+ + ?
show the reaction diagram
Pseudomonas sp. NCIB 9816
-
-
-
-
?
NADH + nitro blue tetrazolium
NAD+ + reduced nitro blue tetrazolium
show the reaction diagram
-
-
-
-
?
NADH + nitro blue tetrazolium
NAD+ + reduced nitro blue tetrazolium
show the reaction diagram
-
-
-
-
?
NADH + nitro blue tetrazolium
NAD+ + reduced nitro blue tetrazolium
show the reaction diagram
Pseudomonas sp. LB400
-
-
-
-
?
NADH + nitro blue tetrazolium
NAD+ + reduced nitro blue tetrazolium
show the reaction diagram
Pseudomonas sp. NCIB 9816
-
-
-
-
?
NADH + nitroblue tetrazolium
NAD+ + reduced nitroblue tetrazolium
show the reaction diagram
-
-
-
-
?
NADPH + cytochrome c
NADP+ + reduced cytochrome c
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. NCIB 9816
-
39% of the activity with NADH
-
-
?
NADPH + H+ + 2,6-dichlorophenolindophenol
NADP+ + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. LB400
-
-
-
-
?
oxidized ferredoxin + NADH
reduced ferredoxin + NAD+
show the reaction diagram
-
-
-
-
?
oxidized ferredoxin + NADH
reduced ferredoxin + NAD+
show the reaction diagram
-
-
-
-
?
oxidized ferredoxin + NADH
reduced ferredoxin + NAD+
show the reaction diagram
-
-
-
-
r
oxidized ferredoxin + NADH
reduced ferredoxin + NAD+
show the reaction diagram
-
-
-
-
?
oxidized ferredoxin + NADH
reduced ferredoxin + NAD+
show the reaction diagram
-
-
-
-
r
oxidized ferredoxin + NADH
reduced ferredoxin + NAD+
show the reaction diagram
-
-
-
-
r
oxidized ferredoxin + NADH
reduced ferredoxin + NAD+
show the reaction diagram
-
ferredoxin from Clostridium tyrobutyricum and Clostridium acetobutylicum
-
-
?
oxidized ferredoxin + NADH
reduced ferredoxin + NAD+
show the reaction diagram
-
the enzyme is required for the mechanism of CO tolerance by the CO-adapted strain
-
-
?
oxidized ferredoxin + NADH
reduced ferredoxin + NAD+
show the reaction diagram
-
the enzyme couples electron flow from formate dehydrogenase (NAD+ requiring) to ferredoxin
-
-
?
oxidized ferredoxin + NADH
reduced ferredoxin + NAD+
show the reaction diagram
-
essential step in glucose fermentation
-
-
?
oxidized ferredoxin + NADH
reduced ferredoxin + NAD+
show the reaction diagram
-
catabolic enzyme
-
-
?
oxidized ferredoxin + NADH
reduced ferredoxin + NAD+
show the reaction diagram
-
required for activity of CYP101
-
-
?
oxidized ferredoxin + NADH
reduced ferredoxin + NAD+
show the reaction diagram
Clostridium tyrobutyricum CNRZ
-
ferredoxin from Clostridium tyrobutyricum and Clostridium acetobutylicum
-
-
?
oxidized ferredoxin + NADPH
reduced ferredoxin + NADP+
show the reaction diagram
-
-
-
-
?
oxidized ferredoxin + NADPH
reduced ferredoxin + NADP+
show the reaction diagram
-
enzyme enhances the level of the active form of CYP105D1
-
-
?
ferredoxin + NADH + H+
reduced ferredoxin + NAD+
show the reaction diagram
-
the enzyme is an essential component of the cytochrome P450 systems in Rhodopseudomonas palustris strain CGA009, a model organism with diverse metabolic pathways, electron transport is initiated by a single two-electron transfer from NAD(P)H to FAD to produce fully reduced FADH2, which provides one electron to the iron-sulfur cluster, in most cases a [2Fe2S] cluster, of ferredoxins. These electrons are finally transferred to the terminal oxygenase, mechanism of electron transport, overview
-
-
?
additional information
?
-
-
the enzyme also has transhydrogenase activity which transfers electrons and protons from NADH to thionicotinamide adenine dinucleotide phosphate and from NADPH to acetylpyridine adenine dinucleotide
-
-
-
additional information
?
-
-
the enzyme can, depending on cellular conditions, produce or oxidize NADH. NADH-ferredoxin reductase can control the level of NAD+ and NADH in the cell
-
-
-
additional information
?
-
-
no substrates of enzyme: morpholine, piperidine, pyrrolidine. Morpholine is a substrate of the P450mor system, consisting of enzyme, Fe3S4 ferredoxin and cytochrome P450
-
-
-
additional information
?
-
Acidovorax sp.
-
the enzyme BphA4 is an FAD-containing NADHdependent ferredoxin reductase
-
-
-
additional information
?
-
-
no substrates of enzyme: morpholine, piperidine, pyrrolidine. Morpholine is a substrate of the P450mor system, consisting of enzyme, Fe3S4 ferredoxin and cytochrome P450
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ferredoxin + NADH + H+
reduced ferredoxin + NAD+
show the reaction diagram
Acidovorax sp.
-
BphA3, a Rieske-type [2Fe-2S] ferredoxin, and BphA4 comprise an electron-transfer system for the multi-component dioxygenase BphA, BphA4 receives two electrons from NADH as a form of hydride and transfers an electron to each of two BphA3 molecules, molecular mechanism of the electron transfer between BphA3 and BphA4, overview
-
-
?
oxidized ferredoxin + NADH
reduced ferredoxin + NAD+
show the reaction diagram
-
-
-
-
?
oxidized ferredoxin + NADH
reduced ferredoxin + NAD+
show the reaction diagram
-
the enzyme is required for the mechanism of CO tolerance by the CO-adapted strain
-
-
?
oxidized ferredoxin + NADH
reduced ferredoxin + NAD+
show the reaction diagram
-
the enzyme couples electron flow from formate dehydrogenase (NAD+ requiring) to ferredoxin
-
-
?
oxidized ferredoxin + NADH
reduced ferredoxin + NAD+
show the reaction diagram
-
essential step in glucose fermentation
-
-
?
oxidized ferredoxin + NADH
reduced ferredoxin + NAD+
show the reaction diagram
-
catabolic enzyme
-
-
?
oxidized ferredoxin + NADH
reduced ferredoxin + NAD+
show the reaction diagram
-
required for activity of CYP101
-
-
?
oxidized ferredoxin + NADPH
reduced ferredoxin + NADP+
show the reaction diagram
-
enzyme enhances the level of the active form of CYP105D1
-
-
?
ferredoxin + NADH + H+
reduced ferredoxin + NAD+
show the reaction diagram
-
the enzyme is an essential component of the cytochrome P450 systems in Rhodopseudomonas palustris strain CGA009, a model organism with diverse metabolic pathways
-
-
?
additional information
?
-
-
the enzyme can, depending on cellular conditions, produce or oxidize NADH. NADH-ferredoxin reductase can control the level of NAD+ and NADH in the cell
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
FAD
-
contains 0.89 mol of FAD per mol of enzyme
FAD
-
1 mol of FAD is bound per mol of enzyme
FAD
-
can bind one mol of FAD per mol of enzyme, Km: 2.5 nM
FAD
-
associated to enzyme, 0.6-0.8 FAD per protein monomer
FAD
-
spectral maxima at 273, 378 and 452 nm, 0.75 mol of FAD per mol of protein
Ferredoxin
Novosphingobium sp.
-
-
-
NADH
-
the CO-adapted strain is a metabolic mutant having higher levels of ferredoxin-NAD+ oxidoreductase activity, which is not inhibited by NADH
NADH
-
preferred cofactor
NADH
Novosphingobium sp.
-
-
NADPH
-
39% of the activity with NADH in the reaction with cytochrome c
NADPH
-
much less effective cofactor than NADH
NADPH
Acidovorax sp.
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Fe2+
-
in an iron-sulfur center
Iron
-
iron-sulfur flavoprotein, protein contains 1.8 gatom of iron and 2.0 gatom of acid-labile sulfur
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1,10-phenanthroline
-
10 mM, 30% inhibition
1,10-phenanthroline
-
-
2,2'-dipyridyl
-
-
iodoacetate
-
10 mM, 50% inhibition
KCl
-
sharp decrease in activity in presence of up to 50 mM KCl, smaller increase in inhibition above
NaCl
-
sharp decrease in activity in presence of up to 50 mM KCl, smaller increase in inhibition above
NADH
-
competitive inhibitor of ferredoxin-NAD+ reductase activity
NADH
-
competitive inhibitor of ferredoxin-NAD+ reductase activity
NADH
-
competitive inhibitor of ferredoxin-NAD+ reductase activity; in cells growing on pyruvate/acetate NADH does not control enzyme activity
NEM
-
10 mM, 67% inhibition
PCMB
-
0.0005 mM, 94% inhibition
Sodium azide
-
40 mM, 46% inhibition
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
acetyl Co-A
-
obligate activator
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0046
-
cytochrome c
-
-
0.0000037
-
Ferredoxin
-
recombinant ferredoxin with C-terminal His-tag, pH 8.6
-
0.0000053
-
Ferredoxin
-
wild-type ferredoxin, pH 8.6
-
0.0105
-
NADH
-
reaction with cytochrome c
0.058
-
NADH
-
reaction with 2,6-dichlorophenolindophenol
0.156
-
NADPH
-
reaction with 2,6-dichlorophenolindophenol
0.0000105
-
Ferredoxin
-
recombinant ferredoxin with N-terminal His-tag, pH 8.6
-
additional information
-
additional information
-
Km-values for transhydrogenase reaction
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
4.5
-
cytochrome c
-
pH 8.5, 30C
60.2
-
ferricyanide
-
pH 8.5, 30C
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.2
-
-
reduction of 2,6-dichlorophenolindophenol
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
32
-
-
reduction of 2,6-dichlorophenolindophenol
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
no activity detected at 5C or at 55C. Activity at 50C is 15.2% of maximal activity
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
34900
-
-
non-denaturing PAGE
36000
-
-
gel filtration
37000
-
-
gel filtration
41500
-
-
NADH-ferredoxin oxidoreductase component of biphenyl 2,3-dioxygenase, gel filtration
46500
-
-
gel filtration
50000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 69000, SDS-PAGE, GST-fusion protein, x * 43000, SDS-PAGE, native protein
?
Novosphingobium sp.
-
x * 39800, recombinant His-tagged RedIIA, SDS-PAGE
?
Novosphingobium sp. KA1
-
x * 39800, recombinant His-tagged RedIIA, SDS-PAGE
-
monomer
-
1 * 43600, NADH-ferredoxin oxidoreductase component of biphenyl 2,3-dioxygenase, SDS-PAGE
monomer
-
1 * 36000, SDS-PAGE
monomer
-
1 * 46000, SDS-PAGE
monomer
-
1 * 42396, calculated, 1 * 50000, SDS-PAGE
monomer
-
dynamic light scattering of PuR
monomer
-
1 * 42396, calculated, 1 * 50000, SDS-PAGE
-
monomer
Pseudomonas sp. LB400
-
1 * 43600, NADH-ferredoxin oxidoreductase component of biphenyl 2,3-dioxygenase, SDS-PAGE
-
monomer
Pseudomonas sp. NCIB 9816
-
1 * 36000, SDS-PAGE
-
monomer
-
dynamic light scattering of PuR
-
additional information
-
enzyme interacts functionally with its redox partner, the Fe3S4 protein ferredoxin, and with the Fe2S2 protein adrenodoxin. Ferredoxin requires its specific NADH:ferredoxin reductase from the P450 system for efficient catalytic function
additional information
Acidovorax sp.
-
BphA3, a Rieske-type [2Fe-2S] ferredoxin, and BphA4 comprise an electron-transfer system for the multi-component dioxygenase BphA
additional information
-
enzyme interacts functionally with its redox partner, the Fe3S4 protein ferredoxin, and with the Fe2S2 protein adrenodoxin. Ferredoxin requires its specific NADH:ferredoxin reductase from the P450 system for efficient catalytic function
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
purified recombinant BphA4 in complex with BphA3, anaerobic crystallization is essential to crystallize the productive complex between oxidized BphA3 and NADH-reduced BphA4, sitting-drop vapour-diffusion method, 26.4 mg/ml BphA4 in 50 mM potassium phosphate buffer, pH 7.0, is reduced with 20 mM NADH for 5 min at 4C, and mixed with oxidized BphA3 at a concentration of 17.5 mg/ml, mixing of 0.9 ml of each of the protein and reservoir solutions, and equilibration against 500 ml reservoir solution, containing 0.2 M ammonium acetate, 0.1 M sodium citrate, pH 5.6, 30% w/v PEG 4000, 20C, X-ray diffraction structure determination and analysis at 1.9 A resolution
Acidovorax sp.
-
recombinant His-tagged RedIIA from KA1, hanging-drop vapour-diffusion, 5C, method using PEG 4000, X-ray diffraction structure determination and analysis at 1.58 A resolution, modelling
Novosphingobium sp.
-
ferredoxin reductase component of biphenyl dioxygenase
-
purified recombinant enzyme, hanging-drop vapour-diffusion method, 0.001 ml of protein solution with 20mg/ml protein in 20 mM HEPES, pH 7.0, and 0.001 ml reservoir solution, containing 100 mM HEPES, pH 7.0-7.2, 16-18% w/v PEG 10 000 at 18C, are mixed and equilibrated against 200 ml reservoir solution, 70 mM sodium cacodylate pH 6.5, 0.98 M sodium acetate, 30% v/v glycerol are best for cryoprotection, X-ray diffraction structure determination and analysis at 2.2 A resolution, molecular replacement using the putidaredoxin reductase structure, overview
-
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.5
-
-
5 d, 30% loss of activity
21
-
-
room temperature, 8 h, 50% loss of activity
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, 3 months, little loss of activity
-
0-4C, stable for up to 30 h
-
-20C, 1 month, minimal loss of activity
-
0-5C, 30% loss of activity after 5 days
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant BphA4 under aerobic conditions from Escherichia coli JM109 cells by anion exchange and adsorption chromatography, and ultrafiltration
Acidovorax sp.
-
recombinant protein
-
recombinant His-tagged RedIIA from Escherichia coli strain BL21(DE3) by metal affinity chromatography and gel filtration
Novosphingobium sp.
-
NADH-ferredoxin oxidoreductase component of biphenyl 2,3-dioxygenase
-
NADH-ferredoxinNAP reductase component of naphthalene dioxygenase
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recombinant enzyme from Escherichia col strain BL21(DE3) by repeated anion exchange chromatography, using a linear gradient of NaCl in 20 mM HEPES buffer, pH 7.0, for elution, followed by gel filtration and ultrafiltration
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Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
overexpression of BphA4 in Escherichia coli JM109 cells
Acidovorax sp.
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expressed in Escherichia coli MV1304 as GST-fusion protein
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gene carAd or fdr, from genomic DNA, expression of His-tagged Red in Escherichia coli strain BL21(DE3)
Novosphingobium sp.
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overexpression of PuR in Escherichia coli strain BL21(DE3) using the tandem tac promoters in the pCWori+ vector
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both isoforms expressed in Escherichia coli BL21 StarTM (DE3) and Streptomyces lividans together with CYP105D1 from Streptomyces griseus
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Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
reconstitution of P450-containing mono-oxygenase using purified homologous enzyme, ferrdoxin and P450
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