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

BRENDA support

Literature summary for 1.1.3.7 extracted from

  • Ferreira, P.; Hernandez-Ortega, A.; Lucas, F.; Carro, J.; Herguedas, B.; Borrelli, K.W.; Guallar, V.; Martinez, A.T.; Medina, M.
    Aromatic stacking interactions govern catalysis in aryl-alcohol oxidase (2015), FEBS J., 282, 3091-3106 .
    View publication on PubMed
Search for more literature for 1.1.3.7

Cloned(Commentary)

Cloned (Comment) Organism
gene aao, recombinant enzyme expression in Escherichia coli Pleurotus eryngii

Protein Variants

Protein Variants Comment Organism
Y92F site-directed mutagenesis, replacement of Tyr92 by phenylalanine does not alter the AAO kinetic constants (on 4-methoxybenzyl alcohol), compared to the wild-type enzyme, most probably because the stacking interaction is still possible Pleurotus eryngii
Y92L site-directed mutagenesis, replacement with a leucine produces a decrease in catalytic efficiency for the alcohol substrate (2.6fold lower), accompanied by approximately twofold increases in both Km(Al) and Kd compared to the wild-type enzyme. The mutation causes a strong decrease in catalytic efficiencies for both O2 (6fold lower) and 4-methoxybenzyl alcohol (860fold lower). As the turnover rate for the Y92W variant is reduced tenfold, the main effect of the mutation concerns the availability of the alcohol substrate at the AAO active site (with 75fold higher Km values). The stacking interactions are strongly affected by this mutation Pleurotus eryngii
Y92W site-directed mutagenesis, introduction of a tryptophan residue at this position only causes a slight increase in KMO2, but strongly reduces the affinity for the substrate (i.e. the pre-steady state Kd and steady-state Km increase by 150fold and 75fold, respectively) and therefore the steady-state catalytic efficiency, compared to the wild-type enzyme, suggesting that proper stacking is impossible with this bulky residue Pleurotus eryngii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information steady-state and stopped-flow kinetics, bi-substrate kinetics analysis, kinetic mechanisms, overview Pleurotus eryngii
0.025
-
 4-methoxybenzyl alcohol pH 6.0, 12°C, recombinant wild-type enzyme Pleurotus eryngii
0.03
-
 4-methoxybenzyl alcohol pH 6.0, 12°C, recombinant mutant Y92F Pleurotus eryngii
0.051
-
 4-methoxybenzyl alcohol pH 6.0, 12°C, recombinant mutant Y92L Pleurotus eryngii
1.6
-
 5-hydroxymethylfurfural pH 6.0, 25°C Pleurotus eryngii
1.89
-
 4-methoxybenzyl alcohol pH 6.0, 12°C, recombinant mutant Y92W Pleurotus eryngii
3.3
-
 2,5-diformylfuran pH 6.0, 25°C Pleurotus eryngii

Localization

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

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2,5-diformylfuran + 2 O2 Pleurotus eryngii
-
 2,5-furandicarboxylic acid + H2O2
-
 ir
5-hydroxymethylfurfural + O2 Pleurotus eryngii
-
 2,5-diformylfuran + H2O2
-
 ?
benzyl alcohol + O2 Pleurotus eryngii
-
 benzaldehyde + H2O2
-
 ?
additional information Pleurotus eryngii the ability of fungal aryl-alcohol oxidase (AAO) to oxidize 5-hydroxymethylfurfural (HMF) results in almost complete conversion into 2,5-formylfurancarboxylic acid (FFCA) in a few hours. The reaction starts with alcohol oxidation, yielding 2,5-diformylfuran (DFF), which is rapidly converted into FFCA by carbonyl oxidation, most probably without leaving the enzyme active site. AAO is combined with an unspecific peroxygenase, UPO, EC 1.11.2.1, from Agrocybe aegerita for full oxidative conversion of 5-hydroxymethylfurfural in an enzymatic cascade. This peroxygenase belongs to the recently described superfamily of hemethiolate peroxidases, and is capable of incorporating peroxide-borne oxygen into diverse substrate molecules. In contrast to AAO, the UPO reaction starts with oxidation of the HMF carbonyl group, yielding 2,5-hydroxymethylfurancarboxylic, which is converted into 2,5-formylfurancarboxylic acid and some 2,5-furandicarboxylic acid ?
-
 ?
additional information Pleurotus eryngii the enzyme typically catalyze the oxidative dehydrogenation of polyunsaturated alcohols using molecular oxygen as the final electron acceptor and producing hydrogen peroxide ?
-
 ?

Organism

Organism UniProt Comment Textmining
Pleurotus eryngii O94219
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme from Escherichia coli Pleurotus eryngii

Reaction

Reaction Comment Organism Reaction ID
an aromatic primary alcohol + O2 = an aromatic aldehyde + H2O2 kinetic and reaction mechanisms involving residue Tyr92, hydride transfer reaction and aromatic stacking interactions on catalysis. Sequential reaction mechanism involving a ternary complex between AAO and its reducing/oxidizing substrates versus a ping-pong mechanism, overview Pleurotus eryngii
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,5-diformylfuran + 2 O2
-
 Pleurotus eryngii 2,5-furandicarboxylic acid + H2O2
-
 ir
3,4-dimethoxybenzyl alcohol + O2 ternary mechanism Pleurotus eryngii 3,4-dimethoxybenzaldehyde + H2O2
-
 ?
3-chloro-4-methoxybenzyl alcohol + O2 ternary mechanism Pleurotus eryngii 3-chloro-4-methoxybenzaldehyde + H2O2
-
 ?
3-chlorobenzyl alcohol + O2 ping-pong mechanism Pleurotus eryngii 3-chlorobenzaldehyde + H2O2
-
 ?
3-fluorobenzyl alcohol + O2 ping-pong mechanism Pleurotus eryngii 3-fluorobenzaldehyde + H2O2
-
 ?
4-methoxybenzyl alcohol + O2 ternary mechanism Pleurotus eryngii 4-methoxybenzaldehyde + H2O2
-
 ?
5-(hydroxymethyl)furan-2-carboxylic acid + O2 very low activity Pleurotus eryngii 2,5-furandicarboxylic acid + ?
-
 ?
5-hydroxymethylfurfural + O2
-
 Pleurotus eryngii 2,5-diformylfuran + H2O2
-
 ?
5-hydroxymethylfurfural + O2
-
 Pleurotus eryngii 5-(hydroxymethyl)furan-2-carboxylic acid + H2O2
-
 ?
benzyl alcohol + O2
-
 Pleurotus eryngii benzaldehyde + H2O2
-
 ?
additional information the ability of fungal aryl-alcohol oxidase (AAO) to oxidize 5-hydroxymethylfurfural (HMF) results in almost complete conversion into 2,5-formylfurancarboxylic acid (FFCA) in a few hours. The reaction starts with alcohol oxidation, yielding 2,5-diformylfuran (DFF), which is rapidly converted into FFCA by carbonyl oxidation, most probably without leaving the enzyme active site. AAO is combined with an unspecific peroxygenase, UPO, EC 1.11.2.1, from Agrocybe aegerita for full oxidative conversion of 5-hydroxymethylfurfural in an enzymatic cascade. This peroxygenase belongs to the recently described superfamily of hemethiolate peroxidases, and is capable of incorporating peroxide-borne oxygen into diverse substrate molecules. In contrast to AAO, the UPO reaction starts with oxidation of the HMF carbonyl group, yielding 2,5-hydroxymethylfurancarboxylic, which is converted into 2,5-formylfurancarboxylic acid and some 2,5-furandicarboxylic acid Pleurotus eryngii ?
-
 ?
additional information the enzyme typically catalyze the oxidative dehydrogenation of polyunsaturated alcohols using molecular oxygen as the final electron acceptor and producing hydrogen peroxide Pleurotus eryngii ?
-
 ?
additional information enzyme AAO is also able to oxidize some furanic compounds such as 5-hydroxymethylfurfural (HMF) and 2,5-diformylfuran (DFF), it has very low activity on 2,5-hydroxymethylfurancarboxylic acid, no activity with 2,5-formylfurancarboxylic acid. NMR analysis of the compounds Pleurotus eryngii ?
-
 ?
additional information the enzyme shows a T-shaped stacking interaction between the Tyr92 side chain and the alcohol substrate at the catalytically competent position for concerted hydride and proton transfers. Bi-substrate kinetics analysis reveals that reactions with 3-chloro- or 3-fluorobenzyl alcohols (halogen substituents) proceed via a ping-pong mechanism. But mono- and dimethoxylated substituents (in 4-methoxybenzyl and 3,4-dimethoxybenzyl alcohols) alter the mechanism and a ternary complex is formed. Stacking energies, reaction mechanism, and kinetic analysis, role of Tyr92 in substrate binding and governing the kinetic mechanism in AAO, overview. Tyr-substrate binding energy and active site structure Pleurotus eryngii ?
-
 ?

Synonyms

Synonyms Comment Organism
AAO
-
 Pleurotus eryngii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
12
-
 steady-state kinetic assays at Pleurotus eryngii
25
-
 assay at Pleurotus eryngii
25
-
 stopped-flow kinetic assays at Pleurotus eryngii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
11
-
 4-methoxybenzyl alcohol pH 6.0, 12°C, recombinant mutant Y92W Pleurotus eryngii
20.1
-
 5-hydroxymethylfurfural pH 6.0, 25°C Pleurotus eryngii
31.4
-
 2,5-diformylfuran pH 6.0, 25°C Pleurotus eryngii
100
-
 4-methoxybenzyl alcohol pH 6.0, 12°C, recombinant mutant Y92L Pleurotus eryngii
120
-
 4-methoxybenzyl alcohol pH 6.0, 12°C, recombinant mutant Y92F Pleurotus eryngii
129
-
 4-methoxybenzyl alcohol pH 6.0, 12°C, recombinant wild-type enzyme Pleurotus eryngii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
-
 Pleurotus eryngii

Cofactor

Cofactor Comment Organism Structure
FAD
-
 Pleurotus eryngii

General Information

General Information Comment Organism
metabolism the enzyme is important in the 5-hydroxymethylfurfural degradation pathway, verview Pleurotus eryngii
physiological function aryl-alcohol oxidase generates H2O2 for lignin degradation at the expense of benzylic and other Pi system-containing primary alcohols, which are oxidized to the corresponding aldehydes Pleurotus eryngii
physiological function the enzyme is involved in lignin degradation. Within this multienzymatic process, which enables the recycling of carbon fixed by photosynthesis in land ecosystems, AAO reduces O2, providing the H2O2 required by ligninolytic peroxidases to oxidize the recalcitrant lignin polymer Pleurotus eryngii

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0167
-
 5-(hydroxymethyl)furan-2-carboxylic acid pH 6.0, 25°C Pleurotus eryngii
0.157
-
 2,5-diformylfuran pH 6.0, 25°C Pleurotus eryngii
0.215
-
 5-hydroxymethylfurfural pH 6.0, 25°C Pleurotus eryngii
6
-
 4-methoxybenzyl alcohol pH 6.0, 12°C, recombinant mutant Y92W Pleurotus eryngii
1940
-
 4-methoxybenzyl alcohol pH 6.0, 12°C, recombinant mutant Y92L Pleurotus eryngii
4450
-
 4-methoxybenzyl alcohol pH 6.0, 12°C, recombinant mutant Y92F Pleurotus eryngii
5160
-
 4-methoxybenzyl alcohol pH 6.0, 12°C, recombinant wild-type enzyme Pleurotus eryngii