1.11.1.16: versatile peroxidase
This is an abbreviated version!
For detailed information about versatile peroxidase, go to the full flat file.
Word Map on EC 1.11.1.16
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1.11.1.16
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lignin
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pleurotus
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peroxidases
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ligninolytic
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eryngii
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laccase
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white-rot
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bjerkandera
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ostreatus
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veratryl
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adusta
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chrysosporium
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phanerochaete
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lignin-degrading
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non-phenolic
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2,6-dimethoxyphenol
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delignification
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aryl-alcohols
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degradation
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synthesis
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industry
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analysis
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paper production
- 1.11.1.16
- lignin
- pleurotus
- peroxidases
-
ligninolytic
- eryngii
- laccase
-
white-rot
- bjerkandera
- ostreatus
-
veratryl
- adusta
- chrysosporium
-
phanerochaete
-
lignin-degrading
-
non-phenolic
- 2,6-dimethoxyphenol
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delignification
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aryl-alcohols
- degradation
- synthesis
- industry
- analysis
- paper production
Reaction
Synonyms
B-type dye-decolorizing peroxidase, bacterial lignin peroxidase, DypB, manganese peroxidase 4, Mb peroxidase, metMb peroxidase, Mnp4, More, myoglobin, R1B4, versatile peroxidase, versatile peroxidase MnP2, versatile peroxidase VPL2 precursor, VP1, Vpl2, VPS1
ECTree
1.11.1.16 versatile peroxidaseAdvanced search results
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results (Engineering
Engineering on EC 1.11.1.16 - versatile peroxidase
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PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
A173R
kcat/KM for Mn2+ is 1.4fold lower than wild-type value, kcat/Km for veratryl alcohol is 1.4fold higher than wild-type value, kcat/Km for Reactive Black 5 is 1.3fold higher than wild-type value
D175A
kcat/KM for Mn2+ is 842fold lower than wild-type value, kcat/Km for veratryl alcohol is3.2 fold higher than wild-type value, kcat/Km for Reactive Black 5 is 1.8fold higher than wild-type value
E140G
substitution of bulky residue at the main heme access channel, kinetic analysis
E140G/K176G
variant attains catalytic efficiencies for oxidation of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) at the heme channel similar to those of the exposed tryptophan site W164
E140G/P141G
substitution of bulky residue at the main heme access channel, kinetic analysis
E140G/P141G/K176G
variant attains catalytic efficiencies for oxidation of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) at the heme channel similar to those of the exposed tryptophan site W164
E140G/P182S/Q229P
site-directed mutagenesis, the mutant BB-8 is active over an enhanced pH range compared to wild-type and displays strong hyperactivation after incubation at alkaline pH with a 3fold increase in activity, The active pH range for mutant BB-8 is expanded considerably for several substrates, including ABTS, sinapic acid and guaiacol. Consequently, BB-8 is active in the acid range (pH 3-4) and remarkably, in the pH interval from 5 to 9 in which the activity of the parental VP is negligible. The kinetic parameters measured for ABTS reveals enhanced catalytic efficiency at acid pH as result of increased affinity, which permits BB-8 to remain active at basic pHs. This effect is mostly attributed to the E140G mutation that enables the mutant to work with similar catalytic efficiency at pH 6 as the parental type at pH 3.5, due to the widening of the heme channel. Whilst the activity against Mn2+ is diminished due to the P182S mutation introduced close to this catalytic site, this mutation offers the first experimental insight into the role of the Mn2+ site for the direct (non-mediated) oxidation of ABTS at neutral/basic pH
E140G/W164S/K176G
variant attains catalytic efficiencies for oxidation of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) at the heme channel similar to those of the exposed tryptophan site W164
E36A
kcat/KM for Mn2+ is 258fold lower than wild-type value, kcat/Km for veratryl alcohol is identical to wild-type value, kcat/Km for Reactive Black 5 is 1.2fold higher than wild-type value
E36A/E40A
kcat/KM for Mn2+ is 16000fold lower than wild-type value, kcat/Km for veratryl alcohol is 1.3fold higher than wild-type value, kcat/Km for Reactive Black 5 is 1.1fold higher than wild-type value
E36A/E40A/D175A
kcat for Mn2+ is 149fold lower than wild-type value, kcat/Km for veratryl alcohol is nearly identical to wild-type value, kcat/Km for Reactive Black 5 is 2fold higher than wild-type value
E36A/E40A/D175A/P327ter
kcat for Mn2+ is 149fold lower than wild-type value, kcat/Km for veratryl alcohol is 1.6fold lower than wild-type value, kcat/Km for Reactive Black 5 is 2.4fold higher than wild-type value
E36D
kcat/KM for Mn2+ is 77fold lower than wild-type value, kcat/Km for veratryl alcohol is 1.3fold higher than wild-type value, kcat/Km for Reactive Black 5 is 3.5fold higher than wild-type value
E37K/H39R/V160A/T184M/Q202L/D213A/G330R
site-directed mutagenesis of enzyme mutant E37K/V160A/T184M/Q202L introducing three additional stabilizing point mutations, the final mutant (2-1B) shows an overall enhancement of 8°C in kinetic thermostability compared to wild-type enzyme, the specific activity increases 2.5fold, and the expression rate is enhanced by 52 fold. The thermostability mutant 2-1B displays remarkable stability at alkaline pH (with a residual activity above 60% at pH 9 after 120 h of incubation), which is rather unusual in fungal peroxidases. Although 2-1B is stable at alkaline conditions, there is hardly any activity at its three catalytic sites at basic pH
E37K/V160A/T184M/Q202L
E40A
kcat/KM for Mn2+ is 1231fold lower than wild-type value, kcat/Km for veratryl alcohol is 1.2fold lower than wild-type value, kcat/Km for Reactive Black 5 is nearly identical to wild-type value
E40D
kcat/KM for Mn2+ is 54fold lower than wild-type value, kcat/Km for veratryl alcohol is 1.3fold lower than wild-type value, kcat/Km for Reactive Black 5 is 2.4fold higher than wild-type value
F142G
substitution of bulky residue at the main heme access channel, kinetic analysis
K176D
substitution of bulky residue at the main heme access channel, kinetic analysis
K176G
substitution of bulky residue at the main heme access channel, kinetic analysis
K215G
substitution of bulky residue at the main heme access channel, kinetic analysis
K215Q
substitution of bulky residue at the main heme access channel, kinetic analysis
N11D/G35K/E40K/T45A/S86R/P141A/F186L/T323I
site-directed mutagenesis
P141G
substitution of bulky residue at the main heme access channel, kinetic analysis
P76G
substitution of bulky residue at the main heme access channel, kinetic analysis
R257A/A260F
R257D
V160I/A260G
site-directed mutagenesis, the mutant MV4 shows increased dye degradation activity compared to the wild-type enzymen with Evans blue, Amido black 10B, and especially with Guinea green B
V160I/A260V
site-directed mutagenesis, the mutant MV5 shows increased dye degradation activity compared to the wild-type enzyme with Evans blue, and Guinea green B, but not with Amido black 10B
V160L/A260S
site-directed mutagenesis, the mutant MV1 shows increased dye degradation activity compared to the wild-type enzyme
V160Y
site-directed mutagenesis, the mutant MV2 shows increased dye degradation activity compared to the wild-type enzyme with Evans blue and Amido black 10B, but not with Guinea green B
V160Y/A260R
site-directed mutagenesis, the mutant MV3 shows increased dye degradation activity compared to the wild-type enzyme with Evans blue and Amido black 10B, but not with Guinea green B
W164H
W164S
W164X
site-directed mutagenesis, no activity at the catalytic Trp164 at basic pH due to the fact that the reduction potential of the Trp164 radical decreases as the pH increases, hindering the oxidation of high-redox potential substrates at neutral/basic pH. The long-range electron transfer pathway from Trp164 to the heme is permanently cancelled out at pHs above pH 5.0, thereby diverting the oxidative route for the oxidation of low-redox potential substrates to the other two catalytic sites at the time that the oxidation of high-redox potential compounds is supressed
W164Y
site-directed mutagenesis, substitution of Trp-164 by a histidine, serine, or tyrosine residues causes a complete loss of activity on veratryl alcohol and Reactive Black 5
W164Y/R257A/A260F
site-directed mutagenesis, substitution of Trp-164 by a histidine, serine, or tyrosine residues causes a complete loss of activity on veratryl alcohol and Reactive Black 5
Q266F
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kinetic properties for H2O2 almost identical to those of wild-type, less than half the RNase A-oxidizing activity of wild-type
R263N
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kinetic properties for H2O2 almost identical to those of wild-type, additional N-glycosylation
V166/168L
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kinetic properties for H2O2 almost identical to those of wild-type
W170A
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kinetic properties for H2O2 almost identical to those of wild-type, no oxidation of veratryl alcohol, decrease in oxodation of RNase A
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
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
additional information
E37K/V160A/T184M/Q202L
mutant obtained by directed evolution, increase in activity and temperature stability
E37K/V160A/T184M/Q202L
site-directed mutagenesis, the secretion of the mutant enzyme from recombinant Saccharomyces cerevisiae improves 129fold compared to wild-type, yielding 22 mg/l of active, soluble and stable enzyme, overexpression in Pichia pastoris, the enzyme is secreted
R257A/A260F
43% decrease in efficiency for oxidizing veratryl alcohol
R257D
versatile peroxidase activity on Reactive Black 5 is eliminated by the R257D mutation
W164H
site-directed mutagenesis, substitution of Trp-164 by a histidine, serine, or tyrosine residues causes a complete loss of activity on veratryl alcohol and Reactive Black 5
W164S
site-directed mutagenesis, substitution of Trp-164 by a histidine, serine, or tyrosine residues causes a complete loss of activity on veratryl alcohol and Reactive Black 5
W164S
loss activity. Residue is responsible for high redox potential substrate oxidation
N246A
mutation inimally 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
additional information
introduction of radical-forming aromatic amino acids by chemical modification of the protein surface is performed using carbodiimide and succiniimide as carboxyl group activators, and the catalytic implications of these additional surface active-sites on the oxidation of 2,6-dimethylphenol, Mn2+, and remazol brilliant blue R (RBBR) are determined. These three different substrates are oxidized in different active sites of the enzyme molecule, of which the high redox RBBR is the only one that is transformed by an external radical formed in the protein surface. Both catalytic constants kcat and KM are significantly affected by the chemical modifications. Tryptophan- and tyrosine-modified versatile peroxidase shows higher catalytic transformation than the unmodified enzyme for RBBR, while the Mn2+ oxidation is significantly reduced by all chemical modifications. Formation of additional protein-based radicals after the chemical modification with radical-forming amino acids is determined by electron paramagnetic resonance studies. The chemical modification could modify any free amino or free carboxylic groups. The access channel to the heme edge is formed by two lysines (K212 and K275) and a glutamic acid (E169) that are prone to be modified. Method overview
additional information
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introduction of radical-forming aromatic amino acids by chemical modification of the protein surface is performed using carbodiimide and succiniimide as carboxyl group activators, and the catalytic implications of these additional surface active-sites on the oxidation of 2,6-dimethylphenol, Mn2+, and remazol brilliant blue R (RBBR) are determined. These three different substrates are oxidized in different active sites of the enzyme molecule, of which the high redox RBBR is the only one that is transformed by an external radical formed in the protein surface. Both catalytic constants kcat and KM are significantly affected by the chemical modifications. Tryptophan- and tyrosine-modified versatile peroxidase shows higher catalytic transformation than the unmodified enzyme for RBBR, while the Mn2+ oxidation is significantly reduced by all chemical modifications. Formation of additional protein-based radicals after the chemical modification with radical-forming amino acids is determined by electron paramagnetic resonance studies. The chemical modification could modify any free amino or free carboxylic groups. The access channel to the heme edge is formed by two lysines (K212 and K275) and a glutamic acid (E169) that are prone to be modified. Method overview
additional information
versatile peroxidase from the fungus Bjerkandera adusta confers abiotic stress tolerance in transgenic tobacco plants. Thirty independent T2 transgenic VP lines overexpressing the fungal Bjerkandera adusta VP gene are selected on kanamycin. The VP22, VP24, and VP27 lines show significant manganese peroxidase (MnP) activity. The highest is VP22, which shows 10.87fold more manganese peroxidase activity than the wild-type plants and leads to a 34% increase in plant height and 28% more biomass. The VP22, VP24, and VP27 lines show enhanced tolerance to drought, 200 mM NaCl, and 400 mM sorbitol. Also, these transgenics display significant tolerance to methyl viologen, an active oxygen-generating compound. The average stem length of transgenic tobacco plants expressing the VP gene is approximately 34% taller than the wild-type plants, and there is a 28% average increase in fresh weight after three months in greenhouse conditions. Phenotypes, detailed overview
additional information
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versatile peroxidase from the fungus Bjerkandera adusta confers abiotic stress tolerance in transgenic tobacco plants. Thirty independent T2 transgenic VP lines overexpressing the fungal Bjerkandera adusta VP gene are selected on kanamycin. The VP22, VP24, and VP27 lines show significant manganese peroxidase (MnP) activity. The highest is VP22, which shows 10.87fold more manganese peroxidase activity than the wild-type plants and leads to a 34% increase in plant height and 28% more biomass. The VP22, VP24, and VP27 lines show enhanced tolerance to drought, 200 mM NaCl, and 400 mM sorbitol. Also, these transgenics display significant tolerance to methyl viologen, an active oxygen-generating compound. The average stem length of transgenic tobacco plants expressing the VP gene is approximately 34% taller than the wild-type plants, and there is a 28% average increase in fresh weight after three months in greenhouse conditions. Phenotypes, detailed overview
additional information
Bjerkandera adusta UAMH8258
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versatile peroxidase from the fungus Bjerkandera adusta confers abiotic stress tolerance in transgenic tobacco plants. Thirty independent T2 transgenic VP lines overexpressing the fungal Bjerkandera adusta VP gene are selected on kanamycin. The VP22, VP24, and VP27 lines show significant manganese peroxidase (MnP) activity. The highest is VP22, which shows 10.87fold more manganese peroxidase activity than the wild-type plants and leads to a 34% increase in plant height and 28% more biomass. The VP22, VP24, and VP27 lines show enhanced tolerance to drought, 200 mM NaCl, and 400 mM sorbitol. Also, these transgenics display significant tolerance to methyl viologen, an active oxygen-generating compound. The average stem length of transgenic tobacco plants expressing the VP gene is approximately 34% taller than the wild-type plants, and there is a 28% average increase in fresh weight after three months in greenhouse conditions. Phenotypes, detailed overview
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additional information
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immobilization of Lentinus squarrosulus on inert polyurethane foam (PUF) with optimized medium for production enhances the versatile peroxidase yield multifold. Maximal yield of versatile peroxidase achieved through optimization and immobilization strategies is 116 U/ml
additional information
Lentinus squarrosulus 12292 ITS
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immobilization of Lentinus squarrosulus on inert polyurethane foam (PUF) with optimized medium for production enhances the versatile peroxidase yield multifold. Maximal yield of versatile peroxidase achieved through optimization and immobilization strategies is 116 U/ml
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additional information
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generation of a recombinant enzyme rVP1 expressed from Escherichia coli in inclusion bodies, renaturation of the recombinant enzyme. Most derivatives of G-type lignin increase slightly by the recombinant enzyme rVP1 treatment compared with the control. The ratio of guaiacyl-type to syringyl-type derivatives (G/S) after rVP1 treatment is 5.4times higher than that of the control. The polymerization of alkali lignin may be attributed to the transformation of S-type into G-type lignin by demethoxylation
additional information
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generation of a recombinant enzyme rVP1 expressed from Escherichia coli in inclusion bodies, renaturation of the recombinant enzyme. Most derivatives of G-type lignin increase slightly by the recombinant enzyme rVP1 treatment compared with the control. The ratio of guaiacyl-type to syringyl-type derivatives (G/S) after rVP1 treatment is 5.4times higher than that of the control. The polymerization of alkali lignin may be attributed to the transformation of S-type into G-type lignin by demethoxylation
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additional information
an engineered N-terminally truncated variant of mutant E37K/V160A/T184M/Q202L displays similar biochemical properties to those of the non-truncated counterpart in terms of kinetics, stability and spectroscopic features. Additional cycles of evolution raised the melting temperature by 8 degrees and significantly increased the enzyme's stability at alkaline pHs. In addition, the Km for H2O2 is enhanced up to 15fold while the catalytic efficiency is maintained, and there is an improvement in peroxide stability
additional information
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an engineered N-terminally truncated variant of mutant E37K/V160A/T184M/Q202L displays similar biochemical properties to those of the non-truncated counterpart in terms of kinetics, stability and spectroscopic features. Additional cycles of evolution raised the melting temperature by 8 degrees and significantly increased the enzyme's stability at alkaline pHs. In addition, the Km for H2O2 is enhanced up to 15fold while the catalytic efficiency is maintained, and there is an improvement in peroxide stability
additional information
due to its broad substrate scope and minor requirements, versatile peroxidase is an extremely attractive blueprint to be designed by the directed evolution tool-box, directed evolution for functional expression in Saccharomyces cerevisiae, directed evolution for activity at alkaline pH, overview
additional information
improvement of degradation of azo dyes by versatile peroxidase through saturation mutagenesis and application in form of VP-coated yeast cell walls. Via saturation mutagenesis, two amino acids in the catalytic tryptophan environment of the enzyme are altered (V160 and A260). Library screening with three azo dyes reveals that these two positions have a significant influence on substrate specificity. Enzyme variants with up to 16fold higher catalytic efficiency for different azo dyes are isolated and sequenced. Immobilization of versatile peroxidase on the surface of yeast cells in purified cell wall fragments after lysis, the enzyme VP embedded in the cell wall retains about 70 % of its initial activity after 10 cycles of dye degradation each lasting 12 h
additional information
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improvement of degradation of azo dyes by versatile peroxidase through saturation mutagenesis and application in form of VP-coated yeast cell walls. Via saturation mutagenesis, two amino acids in the catalytic tryptophan environment of the enzyme are altered (V160 and A260). Library screening with three azo dyes reveals that these two positions have a significant influence on substrate specificity. Enzyme variants with up to 16fold higher catalytic efficiency for different azo dyes are isolated and sequenced. Immobilization of versatile peroxidase on the surface of yeast cells in purified cell wall fragments after lysis, the enzyme VP embedded in the cell wall retains about 70 % of its initial activity after 10 cycles of dye degradation each lasting 12 h
