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Literature summary extracted from

  • Vina-Gonzalez, J.; Elbl, K.; Ponte, X.; Valero, F.; Alcalde, M.
    Functional expression of aryl-alcohol oxidase in Saccharomyces cerevisiae and Pichia pastoris by directed evolution (2018), Biotechnol. Bioeng., 115, 1666-1674 .
    View publication on PubMed

Application

EC Number Application Comment Organism
1.1.3.7 synthesis expression variant carrying 4 mutations in the chimeric signal peptide (prealphaproK), plus mutations H91N/L170M in the mature protein, shows 350fold improved secretion (4.5 mg/l) in Saccharomyces cerevisiae and is stable. Expression in Pichia pastoris and fermentation in a fed-batch bioreactor enhances production to 25 mg/l. Both recombinant AAO from Saccharomyces cerevisiae and Pichia pastoris are subjected to the same N-terminal processing and have a similar pH activity profile, they differ in their kinetic parameters and thermostability Pleurotus eryngii

Protein Variants

EC Number Protein Variants Comment Organism
1.1.3.7 H91N/L170M expression variant carrying 4 mutations in the chimeric signal peptide (prealphaproK), plus mutations H91N/L170M in the mature protein, shows increased secretion upon expression in Pichia pastoris and Saccharomyces cerevisiae Pleurotus eryngii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.3.7 0.022
-
 4-methoxybenzyl alcohol recombinant protein from glycosylation-deficient Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 0.023
-
 4-methoxybenzyl alcohol recombinant protein from wild-type Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 0.037
-
 4-methoxybenzyl alcohol recombinant protein from wild-type Pichia pastoris, pH 6, 25°C Pleurotus eryngii
1.1.3.7 0.095
-
 2,4-hexadien-1-ol recombinant protein from glycosylation-deficient Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 0.096
-
 2,4-hexadien-1-ol recombinant protein from wild-type Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 0.106
-
 2,4-hexadien-1-ol recombinant protein from wild-type Pichia pastoris, pH 6, 25°C Pleurotus eryngii
1.1.3.7 0.34
-
 veratryl alcohol recombinant protein from wild-type Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 0.36
-
 veratryl alcohol recombinant protein from glycosylation-deficient Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 0.37
-
 benzyl alcohol recombinant protein from wild-type Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 0.38
-
 benzyl alcohol recombinant protein from glycosylation-deficient Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 0.41
-
 veratryl alcohol recombinant protein from wild-type Pichia pastoris, pH 6, 25°C Pleurotus eryngii
1.1.3.7 0.44
-
 benzyl alcohol recombinant protein from wild-type Pichia pastoris, pH 6, 25°C Pleurotus eryngii

Organism

EC Number Organism UniProt Comment Textmining
1.1.3.7 Pleurotus eryngii O94219
-
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
1.1.3.7 glycoprotein recombinant protein from wild-type Saccharomyces cerevisiae, 60% glycosylation Pleurotus eryngii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.3.7 2,4-hexadien-1-ol + 2 O2
-
 Pleurotus eryngii 2,4-hexadienal + 2 H2O2
-
 ?
1.1.3.7 4-methoxybenzyl alcohol + O2
-
 Pleurotus eryngii 4-methoxybenzyl aldehyde + H2O2
-
 ?
1.1.3.7 benzyl alcohol + O2
-
 Pleurotus eryngii benzyl aldehyde + H2O2
-
 ?
1.1.3.7 veratryl alcohol + O2
-
 Pleurotus eryngii veratryl aldehyde + H2O2
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.3.7 ? x * 150000, recombinant protein from wild-type Saccharomyces cerevisiae, x * 63000, recombinant protein from glycosylation-deficient Saccharomyces cerevisiae, x * 63000, recombinant protein from wild-type Pichia pastoris Pleurotus eryngii

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.3.7 61.2
-
 melting temperature, recombinant protein from glycosylation-deficient Saccharomyces cerevisiae Pleurotus eryngii
1.1.3.7 61.3
-
 melting temperature, recombinant protein from wild-type Pichia pastoris Pleurotus eryngii
1.1.3.7 63
-
 melting temperature, recombinant protein from wild-type Saccharomyces cerevisiae Pleurotus eryngii

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.3.7 15.1
-
 benzyl alcohol recombinant protein from wild-type Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 23.4
-
 benzyl alcohol recombinant protein from glycosylation-deficient Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 24.6
-
 veratryl alcohol recombinant protein from wild-type Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 34.4
-
 benzyl alcohol recombinant protein from wild-type Pichia pastoris, pH 6, 25°C Pleurotus eryngii
1.1.3.7 36.8
-
 veratryl alcohol recombinant protein from glycosylation-deficient Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 41
-
 4-methoxybenzyl alcohol recombinant protein from wild-type Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 47
-
 2,4-hexadien-1-ol recombinant protein from wild-type Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 56.9
-
 4-methoxybenzyl alcohol recombinant protein from glycosylation-deficient Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 56.9
-
 veratryl alcohol recombinant protein from wild-type Pichia pastoris, pH 6, 25°C Pleurotus eryngii
1.1.3.7 62
-
 2,4-hexadien-1-ol recombinant protein from glycosylation-deficient Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 70
-
 4-methoxybenzyl alcohol recombinant protein from wild-type Pichia pastoris, pH 6, 25°C Pleurotus eryngii
1.1.3.7 89
-
 2,4-hexadien-1-ol recombinant protein from wild-type Pichia pastoris, pH 6, 25°C Pleurotus eryngii

pI Value

EC Number Organism Comment pI Value Maximum pI Value
1.1.3.7 Pleurotus eryngii isoelelctric focusing, recombinant protein from wild-type Pichia pastoris
-
 4.5
1.1.3.7 Pleurotus eryngii isoelectric focusing, recombinant protein from glycosylation-deficient Saccharomyces cerevisiae
-
 4.6
1.1.3.7 Pleurotus eryngii isoelectric focusing, recombinant protein from wild-type Saccharomyces cerevisiae
-
 4.8

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.3.7 41
-
 benzyl alcohol recombinant protein from wild-type Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 62
-
 benzyl alcohol recombinant protein from glycosylation-deficient Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 72
-
 veratryl alcohol recombinant protein from wild-type Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 78
-
 benzyl alcohol recombinant protein from wild-type Pichia pastoris, pH 6, 25°C Pleurotus eryngii
1.1.3.7 102
-
 veratryl alcohol recombinant protein from glycosylation-deficient Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 139
-
 veratryl alcohol recombinant protein from wild-type Pichia pastoris, pH 6, 25°C Pleurotus eryngii
1.1.3.7 456
-
 2,4-hexadien-1-ol recombinant protein from wild-type Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 653
-
 2,4-hexadien-1-ol recombinant protein from glycosylation-deficient Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 840
-
 2,4-hexadien-1-ol recombinant protein from wild-type Pichia pastoris, pH 6, 25°C Pleurotus eryngii
1.1.3.7 1782
-
 4-methoxybenzyl alcohol recombinant protein from wild-type Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii
1.1.3.7 1909
-
 4-methoxybenzyl alcohol recombinant protein from wild-type Pichia pastoris, pH 6, 25°C Pleurotus eryngii
1.1.3.7 2586
-
 4-methoxybenzyl alcohol recombinant protein from glycosylation-deficient Saccharomyces cerevisiae, pH 6, 25°C Pleurotus eryngii