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Information on EC 1.5.1.36 - flavin reductase (NADH)
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1.5.1.36 flavin reductase (NADH)IUBMB Comments
The enzyme from Escherichia coli W catalyses the reduction of free flavins by NADH. The enzyme has similar affinity to FAD, FMN and riboflavin. Activity with NADPH is more than 2 orders of magnitude lower than activity with NADH.
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Word Map
- 1.5.1.36
-
nadph:flavin
- fmnh2
- fischeri
-
alkanesulfonate
-
oxygen-insensitive
-
beneckea
-
desulfonation
-
fmnh2-dependent
- nitroreductases
The enzyme appears in viruses and cellular organisms
Synonyms
fmn reductase, nad(p)h:flavin oxidoreductase, nadh:flavin oxidoreductase, nadh:fmn oxidoreductase, nad(p)h-flavin oxidoreductase, nadh-dependent flavin reductase, fad reductase, flavin mononucleotide reductase, smob-adp1, nadh-flavin oxidoreductase, 
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
BaiH
DszD
FerA
flavin mononucleotide reductase
flavin reductase
flavin:NADH oxidoreductase
FMN reductase
Frd188
HpaC
LJ0548
LJ0549
NAD(P)H-dependent H2O2-forming flavin reductase
NADH-dependent flavin reductase
NADH: flavin oxidoreductase
NADH: flavinoxidore ductase/NADH oxidase
NADH:flavin oxidoreductase
nfr1
nfr2
NOX
Pden2689
HpaC
small component of the 4-hydroxyphenylacetate 3-monooxygenase in Escherichia coli K-12 cells
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
reduced flavin + NAD+ = flavin + NADH + H+
-
-
-
-
reduced flavin + NAD+ = flavin + NADH + H+
FAD reduction by recombinant His10-tagged SMOB-ADP1 follows a random sequential mechanism
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PATHWAY SOURCE
PATHWAYS
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SYSTEMATIC NAME
IUBMB Comments
flavin:NAD+ oxidoreductase
The enzyme from Escherichia coli W catalyses the reduction of free flavins by NADH. The enzyme has similar affinity to FAD, FMN and riboflavin. Activity with NADPH is more than 2 orders of magnitude lower than activity with NADH.
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
FAD + NADH + H+
FADH2 + NAD+
-
HpaC, the small reductase component of the 4-hydroxyphenylacetate 3-monooxygenase. The reductase (HpaC) and the oxygenase component (HpaB) of the monooxygenase are encoded by two different genes. The reductase component uses NADH to catalyze the reduction of a flavin that diffuses to the oxygenase component for oxidation of the substrate by molecular oxygen. HpaC that is recombinantly overproduced in Escherichia coli K12 catalyzes the reduction of free flavins by NADH in preference to NADPH
-
?
FAD + NADH + H+
FADH2 + NAD+
although the HpaC enzyme can also use NADPH as a substrate, its specific activities on FMN, FAD, and riboflavin are more than 2 orders of magnitude lower than those observed in the presence of NADH. Vmax/Km is 37% compared to the value for the reaction of FMN + NADH
-
?
FAD + NADH + H+
FADH2 + NAD+
-
the relative activity with FAD is approximately 31% of that of FMN as the acceptor
-
?
FAD + NADH + H+
FADH2 + NAD+
-
the relative activity with FAD is approximately 31% of that of FMN as the acceptor
-
?
FMN + NADH + H+
FMNH2 + NAD+
-
HpaC, the small reductase component of the 4-hydroxyphenylacetate 3-monooxygenase. The reductase (HpaC) and the oxygenase component (HpaB) of the monooxygenase are encoded by two different genes. The reductase component uses NADH to catalyze the reduction of a flavin that diffuses to the oxygenase component for oxidation of the substrate by molecular oxygen. HpaC that is recombinantly overproduced in Escherichia coli K12 catalyzes the reduction of free flavins by NADH in preference to NADPH
-
?
FMN + NADH + H+
FMNH2 + NAD+
the most effective substrates are NADH and FMN. When FMN is added in a 200fold molar excess of the HpaC protein, it becomes completely reduced, suggesting that the flavin dissociates from the protein and behaves as a true substrate rather than as a tightly bound cofactor. Although the HpaC enzyme can also use NADPH as a substrate, its specific activities on FMN, FAD, and riboflavin are more than 2 orders of magnitude lower than those observed in the presence of NADH
-
?
riboflavin + NADH + H+
reduced riboflavin + NAD+
-
HpaC, the small reductase component of the 4-hydroxyphenylacetate 3-monooxygenase. The reductase (HpaC) and the oxygenase component (HpaB) of the monooxygenase are encoded by two different genes. The reductase component uses NADH to catalyze the reduction of a flavin that diffuses to the oxygenase component for oxidation of the substrate by molecular oxygen. HpaC that is recombinantly overproduced in Escherichia coli K12 catalyzes the reduction of free flavins by NADH in preference to NADPH
-
?
riboflavin + NADH + H+
reduced riboflavin + NAD+
although the HpaC enzyme can also use NADPH as a substrate, its specific activities on FMN, FAD, and riboflavin are more than 2 orders of magnitude lower than those observed in the presence of NADH. Vmax/Km is 70% compared to the value for the reaction of FMN + NADH
-
?
riboflavin + NADH + H+
reduced riboflavin + NAD+
-
-
r
riboflavin + NADH + H+
reduced riboflavin + NAD+
-
-
r
riboflavin + NADH + H+
reduced riboflavin + NAD+
-
the relative activity with riboflavin is approximately 5.8% of that of FMN as the acceptor
-
?
riboflavin + NADH + H+
reduced riboflavin + NAD+
-
the relative activity with riboflavin is approximately 5.8% of that of FMN as the acceptor
-
?
additional information
?
-
-
the enzyme is also active with FMN and NADPH, cf. EC 1.5.1.30
-
?
additional information
?
-
-
the enzyme is also active with FMN and NADPH, cf. EC 1.5.1.30
-
?
additional information
?
-
-
the enzyme supplies reduced FADH2 for other enzymes, e.g. the 4-hydroxylphenylacetate 3-monooxygenase HpaB, which contains bound FADH2 and which protects FADH2 from being oxidized by O2, HpaC binds to HpaB without substrate channeling
-
?
additional information
?
-
no activity with NADPH
-
?
additional information
?
-
no activity with NADPH
-
?
additional information
?
-
the reductase activity requires FMN, flavin adenine dinucleotide (FAD), or riboflavin, that give a similar activity, and is specific for NADH and not NADPH
-
?
additional information
?
-
-
the reductase activity requires FMN, flavin adenine dinucleotide (FAD), or riboflavin, that give a similar activity, and is specific for NADH and not NADPH
-
?
additional information
?
-
the reductase activity requires FMN, flavin adenine dinucleotide (FAD), or riboflavin, that give a similar activity, and is specific for NADH and not NADPH
-
?
additional information
?
-
-
the reductase activity requires FMN, flavin adenine dinucleotide (FAD), or riboflavin, that give a similar activity, and is specific for NADH and not NADPH
-
?
additional information
?
-
-
less than 0.3% of enzyme activity remains when NADPH is used as the electron donor instead of NADH
-
?
additional information
?
-
-
less than 0.3% of enzyme activity remains when NADPH is used as the electron donor instead of NADH
-
?
additional information
?
-
enzyme FerA binds FMN and FAD with comparable affinity in an enthalpically driven, entropically opposed process. The reduced flavin is bound more loosely than the oxidized one, enzyme FerA follows a random-ordered sequence of substrate, NADH and FMN, binding. The primary kinetic isotope effects from stereospecif-ically deuterated nicotinamide nucleotides demonstrate that hydride transfer occurs from the pro-S position and contributes to rate limitation for the overall reaction. Only minor structural changes around Arg106 take place upon FMN binding, role of Arg106 and His146 in binding offlavin and NADH, respectively. Riboflavin (dephosphorylated FMN) also binds to the enzyme
-
?
additional information
?
-
-
enzyme FerA binds FMN and FAD with comparable affinity in an enthalpically driven, entropically opposed process. The reduced flavin is bound more loosely than the oxidized one, enzyme FerA follows a random-ordered sequence of substrate, NADH and FMN, binding. The primary kinetic isotope effects from stereospecif-ically deuterated nicotinamide nucleotides demonstrate that hydride transfer occurs from the pro-S position and contributes to rate limitation for the overall reaction. Only minor structural changes around Arg106 take place upon FMN binding, role of Arg106 and His146 in binding offlavin and NADH, respectively. Riboflavin (dephosphorylated FMN) also binds to the enzyme
-
?
additional information
?
-
enzyme FerA binds FMN and FAD with comparable affinity in an enthalpically driven, entropically opposed process. The reduced flavin is bound more loosely than the oxidized one, enzyme FerA follows a random-ordered sequence of substrate, NADH and FMN, binding. The primary kinetic isotope effects from stereospecif-ically deuterated nicotinamide nucleotides demonstrate that hydride transfer occurs from the pro-S position and contributes to rate limitation for the overall reaction. Only minor structural changes around Arg106 take place upon FMN binding, role of Arg106 and His146 in binding offlavin and NADH, respectively. Riboflavin (dephosphorylated FMN) also binds to the enzyme
-
?
additional information
?
-
-
the enzyme has no measureable activity with NADPH
-
?
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
FAD + NADH + H+
FADH2 + NAD+
-
HpaC, the small reductase component of the 4-hydroxyphenylacetate 3-monooxygenase. The reductase (HpaC) and the oxygenase component (HpaB) of the monooxygenase are encoded by two different genes. The reductase component uses NADH to catalyze the reduction of a flavin that diffuses to the oxygenase component for oxidation of the substrate by molecular oxygen. HpaC that is recombinantly overproduced in Escherichia coli K12 catalyzes the reduction of free flavins by NADH in preference to NADPH
-
-
?
FMN + NADH + H+
FMNH2 + NAD+
-
HpaC, the small reductase component of the 4-hydroxyphenylacetate 3-monooxygenase. The reductase (HpaC) and the oxygenase component (HpaB) of the monooxygenase are encoded by two different genes. The reductase component uses NADH to catalyze the reduction of a flavin that diffuses to the oxygenase component for oxidation of the substrate by molecular oxygen. HpaC that is recombinantly overproduced in Escherichia coli K12 catalyzes the reduction of free flavins by NADH in preference to NADPH
-
-
?
riboflavin + NADH + H+
reduced riboflavin + NAD+
-
HpaC, the small reductase component of the 4-hydroxyphenylacetate 3-monooxygenase. The reductase (HpaC) and the oxygenase component (HpaB) of the monooxygenase are encoded by two different genes. The reductase component uses NADH to catalyze the reduction of a flavin that diffuses to the oxygenase component for oxidation of the substrate by molecular oxygen. HpaC that is recombinantly overproduced in Escherichia coli K12 catalyzes the reduction of free flavins by NADH in preference to NADPH
-
-
?
riboflavin + NADH + H+
reduced riboflavin + NAD+
-
-
-
r
riboflavin + NADH + H+
reduced riboflavin + NAD+
-
-
-
r
additional information
?
-
-
the enzyme supplies reduced FADH2 for other enzymes, e.g. the 4-hydroxylphenylacetate 3-monooxygenase HpaB, which contains bound FADH2 and which protects FADH2 from being oxidized by O2, HpaC binds to HpaB without substrate channeling
-
-
?
additional information
?
-
no activity with NADPH
-
-
?
additional information
?
-
no activity with NADPH
-
-
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
NADH
-
HpaC, the small component of the 4-hydroxyphenylacetate 3-monooxygenase, is recombinantly overproduced in Escherichia coli K12 and catalyzes the reduction of free flavins by NADH in preference to NADPH
NADH
although the HpaC enzyme can also use NADPH as a substrate, its specific activities on FMN, FAD, and riboflavin are more than 2 orders of magnitude lower than those observed in the presence of NADH
NADPH
although the HpaC enzyme can also use NADPH as a substrate, its specific activities on FMN, FAD, and riboflavin are more than 2 orders of magnitude lower than those observed in the presence of NADH
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
no effect on enzyme activity by Cl- and NaN3
additional information
-
the enzyme is not susceptible to Ca2+, Mg2+, Mn2+ and Co2+
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
AMP
mixed noncompetitive versus FMN, and competitive versus NADH
Fe2+
complete inactivation at 0.1 mM, 38% activity remains in presence of 10 mM EDTA
HpaB
-
presence and binding of HpaB reduced the enzyme activity of HpaC due to stabilization and reduced oxidation by O2 of FADH2
-
lumichrome
an FMN analogue completely lacking the ribityl side chain, competitive versus FMN, and mixed noncompetitive versus NADH
styrene oxide
slightly activating at 0.1 mM, inhibitory at 0.2 mM
Acriflavine
0.2 mg/ml, 87% inhibition, acriflavine is a mixture of 3,6-diamino-10-methylacridinium chloride and 3,6-diaminoacridine
additional information
iodoacetate and iodoacetamide show no inhibition
-
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
styrene oxide
slightly activating at 0.1 mM, inhibitory at 0.2 mM
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0031
FAD
pH 7.8, 22°C, the value is obtained with NADH as the electron donor
0.0021
FMN
pH 7.8, 22°C, the value is obtained with NADH as the electron donor
0.04
NADH
pH 7.8, 22°C, the value is obtained with FMN as electron acceptor
0.0026
riboflavin
pH 7.8, 22°C, the value is obtained with NADH as the electron donor
additional information
additional information
Michaelis-Menten kinetics
-
additional information
additional information
-
kinetics, binding studies of HpaC and HpaB
-
additional information
additional information
bi-substrate kinetic analysis, stopped-flow kinetic measurements, detailed overview
-
additional information
additional information
-
bi-substrate kinetic analysis, stopped-flow kinetic measurements, detailed overview
-
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
V(max)/Km-values in 1/min*mg: 33.3 for FMN (with NADH as electron donor), 22.7 for riboflavin (with NADH as electron donor), 12.3 for FAD (with NADH as electron donor)
-
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.2
crude cell extract, about, with FAD, or FMN, or riboflavin, and NADH, pH 7.0, 37°C
0.318
-
strain W-KO, recombinant enzyme Fre, growth on 4-hydroxyphenylacetate as carbon source
25.1
-
strain W-KO, recombinant enzyme HpaC, growth on 4-hydroxyphenylacetate as carbon source
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.8
7.5
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.3 - 7.5
activity irreversibly drops to under 5% at pH 5.3 indicating denaturation
6.5 - 8
the enzyme maintains more than 80% of activity between pH values of 6.5 to 8
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30
37
37
-
assay at, StyB activity increases at temperatures up to 37°C, and decreases thereafter
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pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
genes LJ_0548 and LJ_0549, or nfr1 and nfr2, encoding the subunits 1 and 2 of the enzyme
UniProt
brenda
genes LJ_0548 and LJ_0549, or nfr1 and nfr2, encoding the subunits 1 and 2 of the enzyme
UniProt
brenda
ATCC 11105, genes hpaC and fre encoding the NAD(P)H-flavin oxidoreductase HpaC and the FAD reductase Fre
-
-
brenda
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
additional information
-
enzyme activities with different growth conditions, overview, strain W-KO does not grow on 4-hydroxyphenylacetate as carbon source
brenda
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
SMOB-ADP1 belongs to the flavin reductases of the HpaC-like subfamily. NAD(P)H:flavin oxidoreductase structure comparisons, overview
malfunction
physiological function
additional information
malfunction
deletion of the encoding genes genes nfr1 and nfr2 in Lactobacillus johnsonii leads to a 40fold reduction of hydrogen peroxide formation. H2O2 production in this mutant can only be restored by in trans complementation of both genes
malfunction
inhibition of Nox causes a noticeable decrease in the microsclerotium yields. Silencing of Nox decreases the microsclerotium yield by 98.5%, H2O2 and virulence decrease it by 38% and 21.5%, respectively
malfunction
-
inhibition of Nox causes a noticeable decrease in the microsclerotium yields. Silencing of Nox decreases the microsclerotium yield by 98.5%, H2O2 and virulence decrease it by 38% and 21.5%, respectively
malfunction
-
deletion of the encoding genes genes nfr1 and nfr2 in Lactobacillus johnsonii leads to a 40fold reduction of hydrogen peroxide formation. H2O2 production in this mutant can only be restored by in trans complementation of both genes
physiological function
the 4-hydroxyphenylacetate 3-monooxygenase from Escherichia coli W is a two-component enzyme encoded by the hpaB and hpaC genes and catalyzes the initial reaction in the degradation of 4-hydroxyphenylacetate, i.e., the introduction of a second hydroxyl group into the benzene nucleus at a position ortho to the existing hydroxyl group, giving rise to 3,4-dihydroxyphenylacetate
physiological function
the conserved NADH-dependent flavin reductase is prominently involved in H2O2 production in Lactobacillus johnsonii, overview
physiological function
the enzyme Nox is required for microsclerotium differentiation through regulation of intracellular H2O2 concentration. Additionally Nox has a great impact on the virulence in Nomuraea rileyi in cabbage caterpillar
physiological function
-
the enzyme Nox is required for microsclerotium differentiation through regulation of intracellular H2O2 concentration. Additionally Nox has a great impact on the virulence in Nomuraea rileyi in cabbage caterpillar
physiological function
-
the conserved NADH-dependent flavin reductase is prominently involved in H2O2 production in Lactobacillus johnsonii, overview
additional information
stabilizing effect of another Paracoccus denitrificans protein, the NAD(P)H:acceptor oxidoreducase FerB, against spontaneous oxidation of the FerA-produced dihydroflavin. The turnover rate for NADH oxidation initiated by the addition of FMN is comparable to that for the native, untagged FerA, indicating that the His tag does not interfere with catalysis. Enzyme active ite structure analysis, overview
additional information
-
stabilizing effect of another Paracoccus denitrificans protein, the NAD(P)H:acceptor oxidoreducase FerB, against spontaneous oxidation of the FerA-produced dihydroflavin. The turnover rate for NADH oxidation initiated by the addition of FMN is comparable to that for the native, untagged FerA, indicating that the His tag does not interfere with catalysis. Enzyme active ite structure analysis, overview
additional information
-
stabilizing effect of another Paracoccus denitrificans protein, the NAD(P)H:acceptor oxidoreducase FerB, against spontaneous oxidation of the FerA-produced dihydroflavin. The turnover rate for NADH oxidation initiated by the addition of FMN is comparable to that for the native, untagged FerA, indicating that the His tag does not interfere with catalysis. Enzyme active ite structure analysis, overview
Show AA Sequence and Transmembrane Helices (1952 entries)
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
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PDB
SCOP
CATH
UNIPROT
ORGANISM
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
18522
2 * 18522, small component of the 4-hydroxyphenylacetate 3-monooxygenase, calculated from sequence
18600
-
2 * 18600, HpaC, the recombinant small component of the 4-hydroxyphenylacetate 3-monooxygenase is a homodimer, calculated from sequence
20000
210000
20000
-
2 * 20000, HpaC, the recombinant small component of the 4-hydroxyphenylacetate 3-monooxygenase is a homodimer, SDS-PAGE
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
homodimer
homotrimer
additional information
?
x * 19936.4, recombinant enzyme, mass spectrometry, x * 20000, recombinant enzyme, SDS-PAGE, x * 20080.8, sequence calculation
?
-
x * 19936.4, recombinant enzyme, mass spectrometry, x * 20000, recombinant enzyme, SDS-PAGE, x * 20080.8, sequence calculation
dimer
-
2 * 18600, HpaC, the recombinant small component of the 4-hydroxyphenylacetate 3-monooxygenase is a homodimer, calculated from sequence
dimer
-
2 * 20000, HpaC, the recombinant small component of the 4-hydroxyphenylacetate 3-monooxygenase is a homodimer, SDS-PAGE
homodimer
2 * 22665, His10-tagged enzyme, sequence calculation, 2 * 20000-25000, recombinant His10-tagged enzyme, SDS-PAGE, 2 * 18679, native enzyme, sequence calculation
homodimer
2 * 18522, small component of the 4-hydroxyphenylacetate 3-monooxygenase, calculated from sequence
additional information
overproduction of the small HpaC component in Escherichia coli K-12 cells facilitates the purification of the protein, which is a homodimer that catalyzes the reduction of free flavins by NADH in preference to NADPH
additional information
the two 20-kDa subunit proteins contain flavin mononucleotide (FMN) reductase conserved domains
additional information
-
the two 20-kDa subunit proteins contain flavin mononucleotide (FMN) reductase conserved domains
additional information
-
the two 20-kDa subunit proteins contain flavin mononucleotide (FMN) reductase conserved domains
additional information
crystal structure of FerA reveals a twisted seven-stranded antiparallel beta-barrel, enzyme structure modeling, overview
additional information
-
crystal structure of FerA reveals a twisted seven-stranded antiparallel beta-barrel, enzyme structure modeling, overview
additional information
-
crystal structure of FerA reveals a twisted seven-stranded antiparallel beta-barrel, enzyme structure modeling, overview
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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified recombinant enzyme in apoform or in complex with FMN, sitting drop vapour diffusion method, crystallization from 0.1 M HEPES, pH 7.4, 10% v/v 2-propanol, and 20% w/v PEG 4000, a few days at 20°C, method optimization, X-ray diffraction structure determination and analysis at 1.53-1.85 A resolution, small angle X-ray scattering, molecular replacement method using structure PDB ID 1RZ0 as search model
complexed with FMN, FAD or riboflavin, sitting drop vapor diffusion method, using 1.6 M ammonium sulfate and 4-5% (v/v) isopropanol
-
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
-
improvement of the intracellular environment for enhancing L-arginine production of Corynebacterium glutamicum by inactivation of H2O2-forming flavin reductases and optimization of ATP supply. Construction of mutants of gene frd2, strain 5-5(frd2) and deletion strains 5-5DELTAfrd2 and 5-5DELTAfrd12. The extracellular H2O2 concentrations of mutant 5-5DELTAfrd12 are lower than that of the wild-type strain SYPA5-5, and the extracellular H2O2 concentrations of mutant 5-5(frd2) is increased compared to the wild-type. Flavin reductase activities in frd1 and frd2 overexpression and deletion strains with NADH and FAD, overview
additional information
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improvement of the intracellular environment for enhancing L-arginine production of Corynebacterium glutamicum by inactivation of H2O2-forming flavin reductases and optimization of ATP supply. Construction of mutants of gene frd2, strain 5-5(frd2) and deletion strains 5-5DELTAfrd2 and 5-5DELTAfrd12. The extracellular H2O2 concentrations of mutant 5-5DELTAfrd12 are lower than that of the wild-type strain SYPA5-5, and the extracellular H2O2 concentrations of mutant 5-5(frd2) is increased compared to the wild-type. Flavin reductase activities in frd1 and frd2 overexpression and deletion strains with NADH and FAD, overview
additional information
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construction of an HpaC inactivation mutant strain W-KO
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
45 - 60
-
enzyme activity is stable at temperatures below 45°C. After incubation at 60°C for 30 min, more than 90% of the activity is lost
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, no significant loss of activity is observed during 2 months of storage at this temperature
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation, butyl-Sepharose column chromatography, Source 30Q column chromatography, FMN-Sepharose 6B affinity column chromatography, and Superdex 75 gel filtration
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ammonium sulfate precipitation, HiTrap Q column chromatography, and Superdex 200 gel filtration
native enzyme from strain NCC 533 by ammonium sulfate fractionation, anion exchange chromatography, and gel filtration
recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3)pLysS by nickel affinity chromatography
recombinant His-tagged wild-type enzymes HpaC and Fre from strain BL21(DE3)
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recombinant N-terminally His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
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refolded recombinant enzyme by anion exchange chromatography and gel filtration, or by nickel affinity chromatography and gel filtration, when a His10-tagged enzyme is expressed
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
Escherichia coli strain BL21(DE3)pLysS effectively produces an active and soluble form of StyB as about 9% of the total protein content, when cultivated at 20°C with 0.5 mM IPTG
-
expressed in Escherichia coli BL21(DE3) cells
gene frd2, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant overexpression of N-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3)
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gene Nox, DNA and amino acid sequence determination and analysis, phylogenetic tree, quantitative real-time PCR enzyme expresison analysis
gene Pden_2689, recombinant overexpression of C-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)pLysS
overproduction of the small HpaC component of the 4-hydroxyphenylacetate 3-monooxygenase in Escherichia coli K-12 cells
recombinant overexpression of His-tagged wild-type enzymes HpaC and Fre in strain BL21(DE3), complementation of the inactivation mutant by transient expression of different gene hpaC variants, overview
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sequence comparisons and phylogenetic analysis, cloning in Escherichia coli strain DH5alpha, recombinant expression in Escherichia coli strain BL21(DE3) (pLysS) in inclusion bodies
subcloned into an IPTG-inducible expression vector, pBaiH2.2. Escherichia coli DH5a cells transformed with pBaiH2.2 express 10fold higher levels of NADH:FOR upon induction with IPTG than did Eubacterium sp. VPI 12708 cells induced with cholic acid
the NADH-dependent flavin reductase is encoded by two highly similar genes, LJ_0548 and LJ_0549, or ,nfr1 and nfr2, encoding the subunits 1 and 2 of the enzyme, DNA and amino acid sequence determination and analysis, a plasmid with the promoter region of the LJ_0045 lactate dehydrogenase gene and a bidirectional terminator is used for overexpression of the LJ_0548 and LJ_0549 genes
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
expression level of gene Nox is upregulated and co-related to the intracellular H2O2 concentration during microsclerotium initiation
inducible by cholate
expression level of gene Nox is upregulated and co-related to the intracellular H2O2 concentration during microsclerotium initiation
expression level of gene Nox is upregulated and co-related to the intracellular H2O2 concentration during microsclerotium initiation
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RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
refolding of recombinant enzyme from Escherichia coli strain BL21(DE3) (pLysS) inclusion bodies, with Triton X100 used in the washing buffer, refolding from solubilization buffer containing 8 M urea, 10 mM DTT, 50 mM Tris-HCl, pH 8.0, per mg pellet at 50°C for 30 min, refolding in a 20fold volume of refolding buffer.containing 50 mM Tris–HCl, pH 8.0, 240 mM NaCl, 10 mM KCl, 1 mM Na2EDTA, 1 mM reduced glutathione, 0.1 mM oxidized glutathione, and 0.05 % w/v PEG 4000, followed by ultrafiltration
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Prieto, M.A.; Garcia, J.L.
Molecular characterization of 4-hydroxyphenylacetate 3-hydroxylase of Escherichia coli. A two-protein component enzyme
J. Biol. Chem.
269
22823-22829
1994
Escherichia coli W (Q57501)
brenda
Galan, B.; Diaz, E.; Prieto, M.A.; Garcia, J.L.
Functional analysis of the small component of the 4-hydroxyphenylacetate 3-monooxygenase of Escherichia coli W: a prototype of a new flavin:NAD(P)H reductase subfamily
J. Bacteriol.
182
627-636
2000
Escherichia coli, Escherichia coli W (Q57501)
brenda
Louie, T.M.; Xie, X.S.; Xun, L.
Coordinated production and utilization of FADH2 by NAD(P)H-flavin oxidoreductase and 4-hydroxyphenylacetate 3-monooxygenase
Biochemistry
42
7509-7517
2003
Escherichia coli
brenda
Chakraborty, S.; Ortiz-Maldonado, M.; Entsch, B.; Ballou, D.P.
Studies on the mechanism of p-hydroxyphenylacetate 3-hydroxylase from Pseudomonas aeruginosa: a system composed of a small flavin reductase and a large flavin-dependent oxygenase
Biochemistry
49
372-385
2010
Pseudomonas aeruginosa
brenda
Yeo, Y.J.; Shin, S.; Lee, S.G.; Park, S.; Jeong, Y.J.
Production, purification, and characterization of soluble NADH-flavin Oxidoreductase (StyB) from Pseudomonas putida SN1
J. Microbiol. Biotechnol.
19
362-367
2009
Pseudomonas putida
brenda
Okai, M.; Ohtsuka, J.; Asano, A.; Guo, L.; Miyakawa, T.; Miyazono, K.; Nakamura, A.; Okada, A.; Zheng, H.; Kimura, K.; Nagata, K.; Tanokura, M.
High pressure refolding, purification, and crystallization of flavin reductase from Sulfolobus tokodaii strain 7
Protein Expr. Purif.
84
214-218
2012
Sulfurisphaera tokodaii, Sulfurisphaera tokodaii 7
brenda
Nijvipakul, S.; Ballou, D.P.; Chaiyen, P.
Reduction kinetics of a flavin oxidoreductase LuxG from Photobacterium leiognathi (TH1): half-sites reactivity
Biochemistry
49
9241-9248
2010
Photobacterium leiognathi
brenda
Li, Q.; Feng, J.; Gao, C.; Li, F.; Yu, C.; Meng, L.; Zhang, Z.; Ma, C.; Gu, L.; Wu, G.; Xu, P.
Purification and characterization of a flavin reductase from the biodesulfurizing bacterium Mycobacterium goodii X7B
Process Biochem.
47
1144-1149
2012
Mycolicibacterium goodii, Mycolicibacterium goodii X7B
-
brenda
Hertzberger, R.; Arents, J.; Dekker, H.L.; Pridmore, R.D.; Gysler, C.; Kleerebezem, M.; de Mattos, M.J.
H(2)O(2) production in species of the Lactobacillus acidophilus group a central role for a novel NADH-dependent flavin reductase
Appl. Environ. Microbiol.
80
2229-2239
2014
Lactobacillus acidophilus (Q74HL7), Lactobacillus acidophilus (Q74HL8), Lactobacillus acidophilus, Lactobacillus johnsonii (Q74HL7 AND Q74HL8), Lactobacillus johnsonii, Lactobacillus acidophilus NCC 533 (Q74HL7), Lactobacillus acidophilus NCC 533 (Q74HL8), Lactobacillus johnsonii NCC 533 (Q74HL7 AND Q74HL8), Lactobacillus johnsonii NCC 533
brenda
Groening, J.A.; Kaschabek, S.R.; Schloemann, M.; Tischler, D.
A mechanistic study on SMOB-ADP1 an NADH flavin oxidoreductase of the two-component styrene monooxygenase of Acinetobacter baylyi ADP1
Arch. Microbiol.
196
829-845
2014
Acinetobacter baylyi (Q6F936)
brenda
Man, Z.; Rao, Z.; Xu, M.; Guo, J.; Yang, T.; Zhang, X.; Xu, Z.
Improvement of the intracellular environment for enhancing L-arginine production of Corynebacterium glutamicum by inactivation of H2O2-forming flavin reductases and optimization of ATP supply
Metab. Eng.
38
310-321
2016
Corynebacterium crenatum, Corynebacterium crenatum SYPA5-5
brenda
Sedlacek, V.; Klumpler, T.; Marek, J.; Kucera, I.
Biochemical properties and crystal structure of the flavin reductase FerA from Paracoccus denitrificans
Microbiol. Res.
188-189
9-22
2016
Paracoccus denitrificans (A1B5I2), Paracoccus denitrificans, Paracoccus denitrificans 1222 (A1B5I2)
brenda
Liu, J.; Yin, Y.; Song, Z.; Li, Y.; Jiang, S.; Shao, C.; Wang, Z.
NADH flavin oxidoreductase/NADH oxidase and ROS regulate microsclerotium development in Nomuraea rileyi
World J. Microbiol. Biotechnol.
30
1927-1935
2014
Metarhizium rileyi (A0A059UEY0), Metarhizium rileyi, Metarhizium rileyi CQNr01 (A0A059UEY0)
brenda
Baron, S.; Hylemon, P.
Expression of the bile acid-inducible NADH flavin oxidoreductase gene of Eubacterium sp. VPI 12708 in Escherichia coli
Biochim. Biophys. Acta
1249
145-154
1995
[Clostridium] scindens (P32370), [Clostridium] scindens VPI 12708 (P32370)
brenda
Franklund, C.; Baron, S.; Hylemon, P.
Characterization of the baiH gene encoding a bile acid-inducible NADH flavin oxidoreductase from Eubacterium sp. strain VPI 12708
J. Bacteriol.
175
3002-3012
1993
[Clostridium] scindens (P32370), [Clostridium] scindens VPI 12708 (P32370)
brenda
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