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Literature summary for 1.11.1.16 extracted from

  • Garcia-Ruiz, E.; Gonzalez-Perez, D.; Ruiz-Duenas, F.J.; Martinez, A.T.; Alcalde, M.
    Directed evolution of a temperature-, peroxide- and alkaline pH-tolerant versatile peroxidase (2012), Biochem. J., 441, 487-498.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Saccharomyces cerevisiae Pleurotus eryngii

Protein Variants

Protein Variants Comment Organism
E37K/V160A/T184M/Q202L mutant obtained by directed evolution, increase in activity and temperature stability Pleurotus eryngii
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 Pleurotus eryngii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0066
-
Reactive Black 5 mutant E37K/V160A/T184M/Q202L, pH 3.5, temperature not specified in the publication Pleurotus eryngii
0.007
-
Reactive Black 5 wild-type, pH 3.5, temperature not specified in the publication Pleurotus eryngii
0.045
-
Mn2+ wild-type, pH 3.5, temperature not specified in the publication Pleurotus eryngii
0.051
-
H2O2 wild-type, pH 3.5, temperature not specified in the publication Pleurotus eryngii
0.056
-
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) mutant E37K/V160A/T184M/Q202L, pH 3.5, temperature not specified in the publication Pleurotus eryngii
0.12
-
Mn2+ mutant E37K/V160A/T184M/Q202L, pH 3.5, temperature not specified in the publication Pleurotus eryngii
0.2
-
H2O2 mutant E37K/V160A/T184M/Q202L, pH 3.5, temperature not specified in the publication Pleurotus eryngii
0.54
-
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) wild-type, pH 3.5, temperature not specified in the publication Pleurotus eryngii
6.5
-
2,6-dimethoxyphenol mutant E37K/V160A/T184M/Q202L, pH 3.5, temperature not specified in the publication Pleurotus eryngii
32
-
2,6-dimethoxyphenol wild-type, pH 3.5, temperature not specified in the publication Pleurotus eryngii

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
35244
-
x * 35258, calculated, x * 35244, MALDI-TOF Pleurotus eryngii
35258
-
x * 35258, calculated, x * 35244, MALDI-TOF Pleurotus eryngii

Organism

Organism UniProt Comment Textmining
Pleurotus eryngii O94753 isoform Vpl2
-

Posttranslational Modification

Posttranslational Modification Comment Organism
glycoprotein degree of glycosylation is 9% Pleurotus eryngii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2 2,6-dimethoxyphenol + 2 H2O2
-
Pleurotus eryngii coerulignone + 2 H2O
-
?
2 Mn2+ + 2 H+ + H2O2
-
Pleurotus eryngii 2 Mn3+ + 2 H2O
-
?
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + H2O2
-
Pleurotus eryngii ? + H2O
-
?
Reactive Black 5 + H2O2
-
Pleurotus eryngii oxidized Reactive Black 5 + H2O
-
?

Subunits

Subunits Comment Organism
? x * 35258, calculated, x * 35244, MALDI-TOF Pleurotus eryngii

Synonyms

Synonyms Comment Organism
Vpl2
-
Pleurotus eryngii

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
58
-
wild-type, melting temperature Pleurotus eryngii
59.4
-
mutant E37K/V160A/T184M/Q202L, melting temperature Pleurotus eryngii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
10.6
-
Reactive Black 5 mutant E37K/V160A/T184M/Q202L, pH 3.5, temperature not specified in the publication Pleurotus eryngii
11.8
-
Reactive Black 5 wild-type, pH 3.5, temperature not specified in the publication Pleurotus eryngii
54
-
Mn2+ wild-type, pH 3.5, temperature not specified in the publication Pleurotus eryngii
58
-
2,6-dimethoxyphenol mutant E37K/V160A/T184M/Q202L, pH 3.5, temperature not specified in the publication Pleurotus eryngii
75
-
Mn2+ mutant E37K/V160A/T184M/Q202L, pH 3.5, temperature not specified in the publication Pleurotus eryngii
98
-
2,6-dimethoxyphenol wild-type, pH 3.5, temperature not specified in the publication Pleurotus eryngii
135
-
H2O2 wild-type, pH 3.5, temperature not specified in the publication Pleurotus eryngii
220
-
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) wild-type, pH 3.5, temperature not specified in the publication Pleurotus eryngii
365
-
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) mutant E37K/V160A/T184M/Q202L, pH 3.5, temperature not specified in the publication Pleurotus eryngii
490
-
H2O2 mutant E37K/V160A/T184M/Q202L, pH 3.5, temperature not specified in the publication Pleurotus eryngii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
3
-
wild-type Pleurotus eryngii
3.5
-
mutant E37K/V160A/T184M/Q202L Pleurotus eryngii

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.1
-
2,6-dimethoxyphenol wild-type, pH 3.5, temperature not specified in the publication Pleurotus eryngii
9.1
-
2,6-dimethoxyphenol mutant E37K/V160A/T184M/Q202L, pH 3.5, temperature not specified in the publication Pleurotus eryngii
410
-
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) wild-type, pH 3.5, temperature not specified in the publication Pleurotus eryngii
630
-
Mn2+ mutant E37K/V160A/T184M/Q202L, pH 3.5, temperature not specified in the publication Pleurotus eryngii
1190
-
Mn2+ wild-type, pH 3.5, temperature not specified in the publication Pleurotus eryngii
1600
-
Reactive Black 5 mutant E37K/V160A/T184M/Q202L, pH 3.5, temperature not specified in the publication Pleurotus eryngii
1670
-
Reactive Black 5 wild-type, pH 3.5, temperature not specified in the publication Pleurotus eryngii
2400
-
H2O2 mutant E37K/V160A/T184M/Q202L, pH 3.5, temperature not specified in the publication Pleurotus eryngii
2650
-
H2O2 wild-type, pH 3.5, temperature not specified in the publication Pleurotus eryngii
6480
-
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) mutant E37K/V160A/T184M/Q202L, pH 3.5, temperature not specified in the publication Pleurotus eryngii