Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.11.1.16 extracted from

  • Durdic, K.I.; Ostafe, R.; Durdevic Delmas, A.; Popovic, N.; Schillberg, S.; Fischer, R.; Prodanovic, R.
    Saturation mutagenesis to improve the degradation of azo dyes by versatile peroxidase and application in form of VP-coated yeast cell walls (2020), Enzyme Microb. Technol., 136, 109509 .
    View publication on PubMed

Application

Application Comment Organism
environmental protection the enzyme immobilized on yeast cell wall fragments can be used for longterm bioremediation of environments contaminated with azo dyes Pleurotus eryngii

Cloned(Commentary)

Cloned (Comment) Organism
generation of an enzyme saturation mutagenesis library by site-directed muztagenesis, recombinant expression of enzyme mutants in Saccharomyces cerevisiae strain EBY100, DNA and amino acid sequence determination and analysis.The recombinant enzyme shows microheterogeneity due to hyperglycosylation in Saccharomyces cerevisiae Pleurotus eryngii

Protein Variants

Protein Variants Comment Organism
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 Pleurotus eryngii
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 Pleurotus eryngii
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 Pleurotus eryngii
V160L/A260S site-directed mutagenesis, the mutant MV1 shows increased dye degradation activity compared to the wild-type enzyme Pleurotus eryngii
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 Pleurotus eryngii
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 Pleurotus eryngii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetic parameters for wild-type enzyme and five selected mutant variants with dye substrates, comparisons, overview Pleurotus eryngii

Localization

Localization Comment Organism GeneOntology No. Textmining
extracellular the enzyme is secreted Pleurotus eryngii
-
-

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2 Mn(II) + 2 H+ + H2O2 Pleurotus eryngii
-
2 Mn(III) + 2 H2O
-
?

Organism

Organism UniProt Comment Textmining
Pleurotus eryngii Q9UVP6
-
-

Purification (Commentary)

Purification (Comment) Organism
extraction and purification of Aga2-VP fusion enzyme proteins from Saccharomyces cerevisiae strain EBY100 cell walls by dialysis, and ultrafiltration, followed by ion exchange chromatography Pleurotus eryngii

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
activities of wild-type enzyme and five selected mutant variants with dye substrates, comparisons, overview Pleurotus eryngii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2 Mn(II) + 2 H+ + H2O2
-
Pleurotus eryngii 2 Mn(III) + 2 H2O
-
?
guaiacol + H2O2
-
Pleurotus eryngii oxidized guaiacol + 2 H2O
-
?
additional information the enzyme shows a broad substrate spectrum Pleurotus eryngii ?
-
-
veratryl alcohol + H2O2
-
Pleurotus eryngii 3,4-dimethoxybenzoic acid + 2 H2O
-
?

Subunits

Subunits Comment Organism
? x * 42000, wild-type enzyme, sequence calculation x * 51500, Aga2-fusion enzyme, sequence calculation, x * 50000-65000, recombinant hyperglycosylated Aga2-fusion enzyme, SDS-PAGE Pleurotus eryngii

Synonyms

Synonyms Comment Organism
VPS1 UniProt Pleurotus eryngii
VPS1 gene name, UniProt Pleurotus eryngii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5
-
assay at Pleurotus eryngii

Cofactor

Cofactor Comment Organism Structure
heme
-
Pleurotus eryngii

General Information

General Information Comment Organism
evolution versatile peroxidase (VP) is a lignin-degrading heme-containing oxidoreductase classified as a class II peroxidase, which is secreted by several species of basidiomycetes, mostly from the genera Pleurotus and Bjerkandera Pleurotus eryngii
physiological function versatile peroxidase (VP) from Pleurotus eryngii is a heme-containing peroxidase with a broad substrate spectrum that can break down many structurally distinct pollutants, including azo dyes. Versatile peroxidase is a hybrid enzyme that combines the catalytic characteristics of MnP (i.e., the ability to oxidase Mn2+ to Mn3+, which when complexed by organic acids can oxidize aromatic compounds, EC 1.11.1.13) with the LiP-like ability (EC 1.11.1.14) to use the long-range electron transfer (LRET) pathway based on surface-exposed catalytic tryptophan for the oxidation of compounds with a higher redox potential. Versatile peroxidase can directly oxidize many high-redox-potential dyes, whereas LiP requires various redox mediators to complete the same reaction. Verstaile peroxidase can also oxidize veratryl alcohol but has a much lower affinity for it as compared to lignin peroxidase (LiP) Pleurotus eryngii