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Information on EC 1.11.1.16 - versatile peroxidase

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EC Tree
     1 Oxidoreductases
         1.11 Acting on a peroxide as acceptor
             1.11.1 Peroxidases
                1.11.1.16 versatile peroxidase
IUBMB Comments
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].
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UNIPROT: Q0SE24
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Word Map
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
Synonyms
myoglobin, versatile peroxidase, mb peroxidase, versatile peroxidase mnp2, metmb peroxidase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
B-type dye-decolorizing peroxidase
-
bacterial lignin peroxidase
-
additional information
see also EC 1.11.1.14, cf. EC 1.11.1.19
PATHWAY SOURCE
PATHWAYS
-
-
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
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1-phenyl-1,2-ethandiol + H2O2
? + 2 H2O
show the reaction diagram
substrate of N246A mutant enzyme
-
-
?
1-phenyl-1-propanol + H2O2
? + 2 H2O
show the reaction diagram
substrate of N246A mutant enzyme
-
-
?
1-phenyl-2-propanol + H2O2
? + 2 H2O
show the reaction diagram
substrate of N246A mutant enzyme
-
-
?
1-phenylethanol + H2O2
? + 2 H2O
show the reaction diagram
substrate of N246A mutant enzyme
-
-
?
2 Mn(II) + 2 H+ + H2O2
2 Mn(III) + 2 H2O
show the reaction diagram
activity of EC 1.11.1.13
-
-
?
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + 2 H+ + H2O2
oxidized 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + 2 H2O
show the reaction diagram
-
-
-
?
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + H2O2 + H+
? + H2O
show the reaction diagram
-
-
-
?
2,6-dimethoxyphenol + 2 H+ + H2O2
oxidized 2,6-dimethoxyphenol + 2 H2O
show the reaction diagram
-
-
-
?
Azure B + 2 H+ + H2O2
oxidized Azure B + 2 H2O
show the reaction diagram
dye decolorization, substrate of N246A mutant enzyme
-
-
?
guaiacylglycerol-beta-guaiacyl ether + H2O2
glycerol + guaiacol + 2 H2O
show the reaction diagram
GGE, substrate of mutant enzyme N246A
-
-
?
Reactive Black 5 + 2 H+ + H2O2
oxidized Reactive Black 5 + 2 H2O
show the reaction diagram
dye decolorization, substrate of N246A mutant enzyme
-
-
?
remazol brilliant blue R + H2O2
oxidized remazol brilliant blue R + 2 H2O
show the reaction diagram
substrate of N246A mutant enzyme
-
-
?
veratryl alcohol + H2O2
3,4-dimethoxybenzoic acid + 2 H2O
show the reaction diagram
substrate of N246A mutant enzyme
-
-
?
additional information
?
-
enzyme DypB degrades solvent-obtained fractions of a Kraft lignin. The recombinant mutant enzyme Rh_DypB shows a classical peroxidase activity which is significantly increased by adding Mn2+ ions, kinetic parameters for H2O2, Mn2+, ABTS, and 2,6-DMP are determined. The enzyme shows broad dye-decolorization activity, especially in the presence of Mn2+, oxidizes various aromatic monomers from lignin, and cleaves the guaiacylglycerol-beta-guaiacyl ether (GGE), i.e., the Calpha-Cbeta bond of the dimeric lignin model molecule of beta-O-4 linkages. Under optimized conditions, 2 mM GGE is fully cleaved by recombinant Rh_DypB, generating guaiacol in only 10 min, at a rate of 12.5 micromol/min/mg enzyme. Screening of oxidation activity on monomeric lignin model compounds, overview
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-
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
NaCl
activates 2 fold at 1 M, pH 5.0, 25°C
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
DMSO
50% inhibition at 20%, 90% inhibition at 30%, pH 5.0, 25°C
Tween-80
80% inhibition at 5%, pH 5.0, 25°C
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
14.9 - 15.6
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)
17.5 - 29.5
2,6-dimethoxyphenol
0.015 - 5
H2O2
3.1 - 11.2
Mn(II)
additional information
additional information
Michaelis-Menten kinetics by mutant enzyme N246A
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
12.6 - 15.4
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)
0.45 - 0.5
2,6-dimethoxyphenol
1.8 - 51
H2O2
19 - 21.4
Mn(II)
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.85 - 0.99
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)
0.015 - 0.29
2,6-dimethoxyphenol
4 - 850
H2O2
1.7 - 6.9
Mn(II)
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
recombinant enzyme, in absense of Mn2+
6
recombinant enzyme, in presense of Mn2+
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
55
recombinant enzyme, in absense of Mn2+
65
recombinant enzyme, in presense of Mn2+
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3 - 9
recombinant enzyme, activity range, in absense of Mn2+
47
recombinant enzyme, activity range, in presense of Mn2+
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
Q0SE24_RHOJR
Rhodococcus jostii (strain RHA1)
350
0
37222
TrEMBL
-
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
determination of the secondary structure of the N246Avariant with alpha-helix and beta-strand content of about 29% and about 19%, respectively
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
structural changes in the mutants D153H, D153A, R244L, N246H, N246A, D153A/N246A are confined to the distal heme environment
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D153A
mutation minimally affects the second order rate constant for Compound I formation and the specificity constant for H2O2, but substitution dramatically reduces the stability of Compound I
D153A/N246A
mutation reduces the second order rate constant for Compound I formation and the specificity constant for H2O2 less than 30fold, substitution dramatically reduces the stability of Compound I
D153H
mutant is more than an order of magnitude less reactive with H2O2 than wild-type
N246A
N246H
no detectable peroxidase activity
R244L
mutation abolishes the peroxidase activity, and heme iron of the mutant shows a pH-dependent transition from high spin pH 5 to low spin pH 8.5
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 7
recombinant enzyme, stable at
763987
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 37
the recombinant enzyme maintains a large part of the starting activity following incubation for 24 h in this temperature range
63 - 65
the recombinant enzyme shows a good thermostability with a melting temperature of 63-65°C
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged N246A mutant enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
design of a synthetic gene encoding the N246Avariant of DypB from Rhodococcus jostii strain RHA1 (Rh_DypB) is designed by back translation of the protein sequence, recombinant overexpression of His-tagged N246A mutant enzyme in Escherichia coli strain BL21(DE3), the enzyme is fully produced as folded holoenzyme, thus without the need for a further reconstitution step
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
peroxidases are well-known biocatalysts produced by all organisms, especially microorganisms, and used in a number of biotechnological applications
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Singh, R.; Grigg, J.C.; Armstrong, Z.; Murphy, M.E.; Eltis, L.D.
Distal heme pocket residues of B-type dye-decolorizing peroxidase: arginine but not aspartate is essential for peroxidase activity
J. Biol. Chem.
287
10623-10630
2012
Rhodococcus jostii (Q0SE24)
Manually annotated by BRENDA team
Vignali, E.; Tonin, F.; Pollegioni, L.; Rosini, E.
Characterization and use of a bacterial lignin peroxidase with an improved manganese-oxidative activity
Appl. Microbiol. Biotechnol.
102
10579-10588
2018
Rhodococcus jostii (Q0SE24)
Manually annotated by BRENDA team