Information on EC 1.11.1.16 - versatile peroxidase

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

EC NUMBER
COMMENTARY hide
1.11.1.16
-
RECOMMENDED NAME
GeneOntology No.
versatile peroxidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
1-(4-hydroxy-3-methoxyphenyl)-2-(2-methoxyphenoxy)propane-1,3-diol + H2O2 = 4-hydroxy-3-methoxybenzaldehyde + 2-methoxyphenol + glycolaldehyde + H2O
show the reaction diagram
2 manganese(II) + 2 H+ + H2O2 = 2 manganese(III) + 2 H2O
show the reaction diagram
(2)
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
manganese oxidation I
-
-
SYSTEMATIC NAME
IUBMB Comments
reactive-black-5:hydrogen-peroxide oxidoreductase
A hemoprotein. This ligninolytic peroxidase combines the substrate-specificity characteristics of the two other ligninolytic peroxidases, EC 1.11.1.13, manganese peroxidase and EC 1.11.1.14, lignin peroxidase. Unlike these two enzymes, it is also able to oxidize phenols, hydroquinones and both low- and high-redox-potential dyes, due to a hybrid molecular architecture that involves multiple binding sites for substrates [2,4].
CAS REGISTRY NUMBER
COMMENTARY hide
114995-15-2
-
42613-30-9
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
growth without Mn2+ added, stimulation of enzyme synthesis by addition of glycolate, glyoxylate, or oxalate to medium
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
despite the presence of Mn2+ in the medium, a transformant overexpressing the enzyme produces mnp4 transcripts as well as versatile peroxidase activity as early as 4 days after inoculation. The level of expression is constant throughout 10 days of incubation and the activity is comparable to the typical activity in Mn2+-deficient media
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1,4-benzohydroquinone + H2O2
? + H2O
show the reaction diagram
-
-
-
-
?
1,4-dimethoxybenzene + H2O2
1,4-benzoquinone + H2O
show the reaction diagram
-
Mn2+-independent activity
-
-
?
1-methylanthracene + H2O2
1-methylanthraquinone + H2O
show the reaction diagram
-
at 43% of the rate with 9-methylanthracene
-
-
?
1-naphthol + H2O2
? + H2O
show the reaction diagram
-
-
-
?
2 2,6-dimethoxyphenol + 2 H2O2
coerulignone + 2 H2O
show the reaction diagram
2 2,6-dimethoxyphenol + H2O2
coerulignone + 2 H2O
show the reaction diagram
-
-
-
-
?
2 Mn2+ + 2 H+
2 Mn3+ + H2
show the reaction diagram
-
-
-
-
?
2 Mn2+ + 2 H+ + H2O2
2 Mn3+ + 2 H2O
show the reaction diagram
2 Mn2+ + H2O2 + 2 H+
2 Mn3+ + 2 H2O
show the reaction diagram
-
-
-
-
?
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + H+ + H2O2
?
show the reaction diagram
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + H2O2
? + H2O
show the reaction diagram
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + H2O2 + H+
?
show the reaction diagram
-
-
-
-
?
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + H2O2 + H+
? + H2O
show the reaction diagram
-
-
-
?
2,6-dimethoxybenzohydroquinone + H2O2
? + H2O
show the reaction diagram
-
-
-
-
?
2,6-dimethoxyphenol + H+ + H2O2
?
show the reaction diagram
2,6-dimethoxyphenol + H2O2
?
show the reaction diagram
-
-
-
-
?
2,6-dimethoxyphenol + H2O2
coerulignone + ?
show the reaction diagram
2,7-diaminofluorene + H2O2
? + H2O
show the reaction diagram
-
during oxidation of 2,7-diaminofluorene, both with and without Mn2+, biphasic kinetics with apparent saturation in both micromolar and millimolar ranges are obtained
-
-
?
2-chloro-1,4-dimethoxybenzene + H2O2
2-chloro-1,4-benzoquinone + H2O
show the reaction diagram
-
Mn2+-independent activity
-
-
?
2-methoxy-1,4-benzohydroquinone + H2O2
? + H2O
show the reaction diagram
-
-
-
-
?
2-methylanthracene + H2O2
2-methylanthraquinone + H2O
show the reaction diagram
-
at 24% of the rate with 9-methylanthracene
-
-
?
3-hydroxyanthranilic acid + H2O2
? + H2O
show the reaction diagram
-
Mn2+-independent activity
-
-
?
3-methyl-2-benzothiazolinone hydrazone + H2O2
? + H2O
show the reaction diagram
-
enzyme has several substrate binding sites for 3-methyl-2-benzothiazolinone hydrazone, in addition to low and high affinity binding sites for Mn2+
-
-
?
4-aminobenzoic acid + H2O2
? + H2O
show the reaction diagram
-
-
-
-
?
4-hydroquinone + H2O2
4-benzoquinone + H2O
show the reaction diagram
-
-
-
-
?
9-methylanthracene + H2O2
10-methylanthracene-9-one + H2O
show the reaction diagram
-
-
-
-
?
acetosyringone + H2O2 + H+
oxidized acetosyringone + H2O
show the reaction diagram
-
-
-
-
?
Acid Blue 62 + H2O2 + H+
oxidized Acid Blue 62 + H2O
show the reaction diagram
-
-
-
-
?
amaranth + H2O2
? + H2O
show the reaction diagram
-
-
-
?
anthracene + H2O2
9,10-anthraquinone + H2O
show the reaction diagram
-
-
-
-
?
anthracene + H2O2
anthraquinone + H2O
show the reaction diagram
-
at 4.8% of the rate with 9-methylanthracene
-
-
?
beta-carotene + H+ + H2O2
?
show the reaction diagram
-
-
-
?
bovine pancreatic RNase
oxidized bovine pancreatic RNase
show the reaction diagram
-
no redox mediators involved
-
-
?
carbazole + H2O2
? + H2O
show the reaction diagram
-
at 4.8% of the rate with 9-methylanthracene
-
-
?
catechol + H2O2
2-benzoquinone + H2O
show the reaction diagram
-
-
-
-
?
chrysene + H2O2
? + H2O
show the reaction diagram
-
-
-
-
?
fluoranthene + H2O2
? + H2O
show the reaction diagram
-
-
-
-
?
fluorene + H2O2
9-fluorenone + H2O
show the reaction diagram
-
-
-
-
?
fulvic acid + H2O2
? + H2O
show the reaction diagram
-
-
-
-
?
guaiacol + H2O2
3,3'-dimethoxy-4,4'-biphenylquinone + H2O
show the reaction diagram
-
-
-
-
?
guaiacol + H2O2
? + H2O
show the reaction diagram
guaiacol + H2O2 + H+
oxidized guaiacol + H2O
show the reaction diagram
humic acid + H2O2
? + H2O
show the reaction diagram
-
-
-
-
?
lignin + H2O2
? + H2O
show the reaction diagram
manganese(II)-substituted polyoxometalate + H2O2
manganese(III)-substituted polyoxometalate + H2O2
show the reaction diagram
-
-
-
-
?
methoxyhydroquinone + H2O2
? + H2O
show the reaction diagram
methylene blue + H2O2
? + H2O
show the reaction diagram
-
-
-
-
?
Mn2+ + H+ + H2O2
Mn3+ + H2O
show the reaction diagram
Mn2+ + H2O2
?
show the reaction diagram
-
-
-
-
?
Mn2+ + H2O2
Mn3+ + H2O
show the reaction diagram
Mn2+ + H2O2 + 2,6-dimethoxyphenol
?
show the reaction diagram
-
-
-
-
?
Mn2+ + H2O2 + guaiacol
?
show the reaction diagram
-
-
-
-
?
Mn2+ + H2O2 + phenol red
?
show the reaction diagram
-
-
-
-
?
Mn2+ + H2O2 + Reactive Black 5
?
show the reaction diagram
-
-
-
-
?
Mn2+ + H2O2 + remazol black-5
?
show the reaction diagram
-
incubation of enzyme with dyes rose bengal, remazol brilliant violet, remazol black-5, remazol blue-19, and remazol orange-16, results in the decolorization of all the dyes tested within a range of 71-84% after 16 h incubation with the enzyme at 100 U/l
-
-
-
Mn2+ + H2O2 + remazol blue-19
?
show the reaction diagram
-
incubation of enzyme with dyes rose bengal, remazol brilliant violet, remazol black-5, remazol blue-19, and remazol orange-16, results in the decolorization of all the dyes tested within a range of 71-84% after 16 h incubation with the enzyme at 100 U/l
-
-
-
Mn2+ + H2O2 + remazol brilliant violet
?
show the reaction diagram
-
incubation of enzyme with dyes rose bengal, remazol brilliant violet, remazol black-5, remazol blue-19, and remazol orange-16, results in the decolorization of all the dyes tested within a range of 71-84% after 16 h incubation with the enzyme at 100 U/l
-
-
-
Mn2+ + H2O2 + remazol orange-16
?
show the reaction diagram
-
incubation of enzyme with dyes rose bengal, remazol brilliant violet, remazol black-5, remazol blue-19, and remazol orange-16, results in the decolorization of all the dyes tested within a range of 71-84% after 16 h incubation with the enzyme at 100 U/l
-
-
-
Mn2+ + H2O2 + rose bengal
?
show the reaction diagram
-
incubation of enzyme with dyes rose bengal, remazol brilliant violet, remazol black-5, remazol blue-19, and remazol orange-16, results in the decolorization of all the dyes tested within a range of 71-84% after 16 h incubation with the enzyme at 100 U/l
-
-
-
Mn2+ + H2O2 + veratryl alcohol
?
show the reaction diagram
-
-
-
-
?
Mordant Black 9 + H2O2 + H+
oxidized Mordant Black 9 + H2O
show the reaction diagram
-
-
-
-
?
NADH + H2O2
NAD+ + H2O
show the reaction diagram
-
-
-
-
?
o-anisidine + H2O2
? + H2O
show the reaction diagram
-
Mn2+-independent activity
-
-
?
Orange II + H+ + H2O2
?
show the reaction diagram
Orange II + H2O2
? + H2O
show the reaction diagram
-
-
-
?
p-anisidine + H2O2
? + H2O
show the reaction diagram
-
Mn2+-independent activity
-
-
?
p-dimethoxybenzene + H2O2
benzoquinone + H2O
show the reaction diagram
-
catalyzed by isoforms PS3, PS1
-
-
?
phenanthrene + H2O2
9,10-phenanthrenequinone + H2O
show the reaction diagram
-
-
-
-
?
Phenol Red + H+ + H2O2
?
show the reaction diagram
phenol red + H2O2
oxidized phenol red + H2O
show the reaction diagram
-
Mn2+-dependent activity
-
-
?
Poly R-478
oxidized Poly R-478
show the reaction diagram
-
no redox mediators involved
-
-
?
Poly R-478 + H2O2
?
show the reaction diagram
-
decolorization of the dye
-
-
?
pyrene + H2O2
? + H2O
show the reaction diagram
-
-
-
-
?
Reactive Black 5 + H+ + H2O2
?
show the reaction diagram
Reactive Black 5 + H2O2
?
show the reaction diagram
Reactive Black 5 + H2O2
? + H2O
show the reaction diagram
Reactive Black 5 + H2O2
oxidized Reactive Black 5 + H2O
show the reaction diagram
Reactive Blue 38 + H2O2
? + H2O
show the reaction diagram
-
-
-
-
?
Reactive Blue 5 + H2O2 + H+
oxidized Reactive Blue 5 + H2O
show the reaction diagram
Reactive Blue 72 + H2O2
? + H2O
show the reaction diagram
-
-
-
-
?
Reactive Violet 5 + H2O2
? + H2O
show the reaction diagram
-
-
-
-
?
RNase A + H2O2
?
show the reaction diagram
-
-
-
-
?
syringaldazine + H2O2
?
show the reaction diagram
-
-
-
-
?
syringaldehyde + H2O2 + H+
oxidized syringaldehyde + H2O
show the reaction diagram
syringol + H+ + H2O2
?
show the reaction diagram
-
-
-
?
syringol + H2O2
? + H2O
show the reaction diagram
-
-
-
-
?
vanillylidenacetone + H2O2
? + H2O
show the reaction diagram
-
Mn2+-dependent activity
-
-
?
veratryl alcohol + H+ + H2O2
veratraldehyde + H2O
show the reaction diagram
-
-
-
?
veratryl alcohol + H2O2 + H+
veratraldehyde + H2O
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
Mn2+ + H+ + H2O2
Mn3+ + H2O
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Iron
-
at pH 4.5, Ca2+-depleted enzyme has a high-spin Fe3+
Manganese
Zn2+
-
5 mM, 108% of initial activity
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)
-
substrate inhibition
bovine pancreatic RNase
-
competitive to oxidation of veratryl alcohol, non-competitive to oxidation of Mn2+
-
Co2+
-
5 mM, 80% of initial activity
Cu2+
-
5 mM, 18% of initial activity
Fe2+
-
5 mM, 11% of initial activity
Mg2+
-
5 mM, 85% of initial activity
Mn2+
-
above 0.1 mM, severe inhibition of oxidation of veratryl alcohol
N-bromosuccinimide
-
modification of tryptophan residues by N-bromosuccinimide drastically reduces the Mn(II)-independent activity and dye decoloration, while Mn(II)-dependent activity is maintained. Effect is not reversed by addition of mediators
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
70
1-naphthol
pH 2.5, 30°C
0.0007 - 2.86
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)
0.01 - 76
2,6-dimethoxyphenol
0.009 - 0.038
2,7-diaminofluorene
0.0059 - 2.24
4-hydroquinone
0.035
acetosyringone
-
pH 5, 25°C
0.03
Acid Blue 62
-
pH 5, 25°C
0.05
beta-carotene
at 30°C and pH 4.5
0.034 - 10.5
catechol
0.035 - 39.8
guaiacol
0.002 - 5
H2O2
6.4
manganese(II)-substituted polyoxometalate
-
in 0.1 M sodium tartrate, pH 5.0, at 20°C
-
0.017 - 3
methoxyhydroquinone
0.007 - 0.013
methylene blue
0.012 - 76.4
Mn2+
0.32
Mordant black 9
-
pH 5, 25°C
0.0358
Orange II
-
at pH 3.0 and 25°C
2.4
p-dimethoxybenzene
-
pH 3.0, isoenzyme PS1
0.008
Phenol red
-
pH 4.5, 30°C
0.0013 - 0.0248
Reactive Black 5
0.089
Reactive Blue 38
-
pH 4.0
0.04
Reactive Blue 5
-
pH 5, 25°C
0.027
Reactive Blue 72
-
pH 4.0
0.047
Reactive Violet 5
-
pH 4.0
0.035 - 0.066
syringaldazine
0.048
syringaldehyde
-
pH 5, 25°C
0.14 - 1
syringol
0.005
vanillylidenacetone
-
presence of Mn2+, isoenzyme MP-1, pH 5.0; presence of Mn2+, isoenzyme MP-2, pH 5.0
0.116 - 54.7
veratryl alcohol
additional information
fulvic acid
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.7 - 12300
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)
2.3 - 293
2,6-dimethoxyphenol
0.19 - 0.83
2,7-diaminofluorene
2.4
2-methylanthracene
-
pH 4.0
7.2 - 108
4-hydroquinone
1000
beta-carotene
at 30°C and pH 4.5
8 - 185.6
catechol
0.018 - 0.045
fulvic acid
9.3 - 185.6
guaiacol
0.6 - 490
H2O2
0.02 - 0.021
humic acid
47
manganese(II)-substituted polyoxometalate
-
in 0.1 M sodium tartrate, pH 5.0, at 20°C
-
4 - 19
methoxyhydroquinone
0.18 - 2.92
methylene blue
2 - 467
Mn2+
0.83
Orange II
-
at pH 3.0 and 25°C
4
p-dimethoxybenzene
-
pH 3.0, isoenzyme PS1
0.4 - 54
Reactive Black 5
19.8
Reactive Blue 38
-
pH 4.0
10
Reactive Blue 72
-
pH 4.0
16.9
Reactive Violet 5
-
pH 4.0
0.22 - 6.1
syringaldazine
3 - 54500
syringol
1.4 - 1600
veratryl alcohol
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1 - 6480
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)
1.3 - 476.2
2,6-dimethoxyphenol
21.1 - 21.8
2,7-diaminofluorene
8.9 - 1600
4-hydroquinone
2.4
acetosyringone
-
pH 5, 25°C
240
Acid Blue 62
-
pH 5, 25°C
1 - 70.7
catechol
0.6 - 17.6
guaiacol
4 - 5961
H2O2
7.36
manganese(II)-substituted polyoxometalate
-
in 0.1 M sodium tartrate, pH 5.0, at 20°C
-
5.7 - 2850
Mn2+
50
Mordant black 9
-
pH 5, 25°C
15.7
Orange II
-
at pH 3.0 and 25°C
1600 - 2224.4
Reactive Black 5
200
Reactive Blue 5
-
pH 5, 25°C
3.3 - 174.3
syringaldazine
1.2
syringaldehyde
-
pH 5, 25°C
1.3 - 50.6
veratryl alcohol
additional information
fulvic acid
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.4
with beta-carotene as substrate, at 30°C and pH 4.5
1.2
with syringol as substrate, at 30°C and pH 4.5
1.4
with 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) as substrate, at 30°C and pH 4.5
2.2
-
substrate Reactive Black 5, pH 3.5, 25°C
2.3
with veratryl alcohol as substrate, at 30°C and pH 4.5
7.2
-
substrate veratryl alcohol, pH 3.0, 25°C
8.4
-
substrate 1,4-benzohydroquinone, pH 5.0, absence of Mn2+
8.8
-
substrate 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), pH 3.5, 25°C
9.6
-
substrate 2-methoxy-1,4-benzohydroquinone, pH 5.0, absence of Mn2+
10.8
-
substrate 2,6-dimthoxy-1,4-benzohydroquinone, pH 5.0, absence of Mn2+
10.86
-
unpurified enzyme, with Mn2+ as substrate, at pH 5.0 and 25°C
26.6
-
pH 4.5, 30°C
61.48
-
after 5.66fold purification, with Mn2+ as substrate, at pH 5.0 and 25°C
80
-
pH 4.5
194
-
substrate Mn2+, pH 5.0, 25°C
334
-
growth on rich peptone medium containing 0.5 mM Mn2+
559
-
after growth on rich peptone medium
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3 - 3.5
-
lignin peroxidase activity in absence of Mn2+
4.2
-
and 3.0, substrate 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), presence of Mn2+
6.5
-
and 3.0, substrate veratryl alcohol, absence of Mn2+; and 3.0, substrate veratryl alcohol, presence of Mn2+; and 5.0, substrate 2,7-diaminofluorene, presence of Mn2+
7
-
and 4.5, substrate 2,7-diaminofluorene, absence of Mn2+; substrate syringaldazine, absence and presence of Mn2+; substrate syringaldazine, absence of Mn2+
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3 - 6.5
more than 60% activity between pH 3.0 and 6.5
4.5 - 6
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40
-
lignin peroxidase activity in absence of Mn2+
60
-
manganese peroxidase activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.5
-
isoelectric focusing
3.6
isoelectric focusing
3.67
-
isoenzyme PS1
3.75
-
isoelectric focusing, isoenzyme MP-2
3.8
-
isoenzyme PS3
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
32000
-
4 * 32000, SDS-PAGE
35244
-
x * 35258, calculated, x * 35244, MALDI-TOF
35258
-
x * 35258, calculated, x * 35244, MALDI-TOF
36400
x * 36400, calculated
40000
-
x * 40000, SDS-PAGE
41000
x * 43000, SDS-PAGE, x * 41000, SDS-PAGE of deglycosylated enzyme
42000
-
x * 45000, isoenzymes PS1, PS2, x * 42000, isoenzyme PS3
45000
-
x * 45000, isoenzymes PS1, PS2, x * 42000, isoenzyme PS3
47000
-
x * 47000, SDS-PAGE
120000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
tetramer
additional information
-
W170 exposed on enzyme surface is a substrate-binding site both for veratryl alcohol and for polymeric substrates
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
construction of enzyme model and identification of active sites for oxidation of Mn2+ and of aromatic substrates
hanging drop vapor diffusion method, crystal structures of untreated versatile peroxidase (immediately after expression in Escherichia coli and in vitro reconstitution), native versatile peroxidase (treated with Mn2+), D175A variant, and wild-type verstile peroxidase (from Pleurotus eryngii culture)
mutant enzyme W164Y, sitting-drop vapor diffusion method, resolution 1.94 A
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mutants E140G, P141G, K176G, and E140G/K176G, to 1.6, 2.0, 1.5, 1.7 and 2.35 A resolution, respectively
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sitting drop vapor diffusion method, using 0.1 M sodium MES buffer at pH 6.5, 25% (w/v) PEG 4000 and 0.2 M MgCl2
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wild-type and mutant M247F
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resonance Raman and electrochemical study. In solution, enzyme shows a heterogeneous spin population, with the five-coordinated quantum mechanically mixed-spin state being the most populated in the latter. The spin population is sensitively dependent on the pH, temperature, and physical, i.e., solution versus crystal versus immobilized, state of the enzymes. The redox potential for the Fe2+/Fe3+ couple is -260 mV
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structural changes in the mutants D153H, D153A, R244L, N246H, N246A, D153A/N246A are confined to the distal heme environment
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 55
-
Residual activity of roughly 100% is found after incubation at 25 to 55°C with all three substrates. Stability decreases gradually at temperatures from 60 to 70°C
35
-
pH 4.5, stable for 1 h
58
-
wild-type, melting temperature
59.4
-
mutant E37K/V160A/T184M/Q202L, melting temperature
79
melting temperature
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, stable
-
4°C, pH 4-7, wild-type is stable for 96 h
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4°C, stable for at least 72h
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation, HiTrap Q column chromatography, and Mono Q column chromatography
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DEAE FF' column chromatography and Superdex 200 gel filtration
HiTrap Sepharose column chromatography
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recombinant protein
resource-Q chromatography
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Resource-Q column chromatography
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