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Literature summary for 1.13.11.60 extracted from

  • Brodhun, F.; Goebel, C.; Hornung, E.; Feussner, I.
    Identification of PpoA from Aspergillus nidulans as a fusion protein of a fatty acid heme dioxygenase/peroxidase and a cytochrome P450 (2009), J. Biol. Chem., 284, 11792-11805.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Aspergillus nidulans

Protein Variants

Protein Variants Comment Organism
H1004A mutant enzyme with abolished 8-hydroperoxide isomerase activity, whereas dioxygenase activity is still present Aspergillus nidulans
Y374 the mutant enzyme shows no detectable activity when incubated with (9Z,12Z)-octadeca-9,12-dienoate as a substrate. When incubated with the intermediate product (8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate it is able to catalyze isomerization to (5S,8R,9Z,12Z)-5,8-dihydroperoxy-9,12-octadecadienoate Aspergillus nidulans

Inhibitors

Inhibitors Comment Organism Structure
additional information suicide inactivation, activity falls to zero within a few min although substrates were available in sufficient amounts Aspergillus nidulans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.005
-
palmitoleate pH 7.2, 24°C Aspergillus nidulans
0.00671
-
oleate pH 7.2, 24°C Aspergillus nidulans
0.0183
-
linoleate pH 7.2, 24°C Aspergillus nidulans
0.0226
-
(11Z)-icos-11-enoic acid pH 7.2, 24°C Aspergillus nidulans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
110000
-
4 * 110000, SDS-PAGE Aspergillus nidulans
440000
-
gel filtration Aspergillus nidulans

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
linoleate + O2 Aspergillus nidulans the enzyme is involved in the regulation of the life cycle of Aspergillus nidulans. Synthesis of the psi factor (8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate, that influences the development of the asexual conidiophores and sexual cleistothecia (8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
-
?

Organism

Organism UniProt Comment Textmining
Aspergillus nidulans Q6RET3
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Aspergillus nidulans

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2
-
pH 7.2, 24°C Aspergillus nidulans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(11Z)-icos-11-enoic acid + O2
-
Aspergillus nidulans 10-hydroperoxyeicosenoic acid
-
?
(11Z,14Z)-icosa-11,14-dienoic acid + O2
-
Aspergillus nidulans (11Z,14Z)-10-hydroperoxyeicosa-11,14-dienoic acid
-
?
(11Z,14Z,17Z)-icosa-11,14,17-trienoic acid + O2
-
Aspergillus nidulans (11Z,14Z,17Z)-10-hydroperoxyeicosa-11,14,17-trienoic acid
-
?
linoleate + O2 the enzyme is involved in the regulation of the life cycle of Aspergillus nidulans. Synthesis of the psi factor (8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate, that influences the development of the asexual conidiophores and sexual cleistothecia Aspergillus nidulans (8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
-
?
linoleate + O2 i.e. (9Z,12Z)-octadeca-9,12-dienoate. The N-terminal heme peroxidase domain might be responsible for the dioxygenase reaction as the first step of the PpoA reaction, i.e. oxidation of linoleic acid to (8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate as an intermediate product. The C-terminal P450 domain catalyzes the second reaction step, the isomerization of (8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate to (5S,8R,9Z,12Z)-5,8-dihydroperoxy-9,12-octadecadienoate, and is therefore termed the 8-hydroperoxide isomerase P450 domain Aspergillus nidulans (8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate
-
?
oleate + O2
-
Aspergillus nidulans (9Z)-8-hydroperoxyoctadec-9-enoate
-
?
palmitoleate + O2
-
Aspergillus nidulans (9Z)-8-hydroperoxyhexadec-9-enoate
-
?

Subunits

Subunits Comment Organism
tetramer 4 * 110000, SDS-PAGE Aspergillus nidulans

Synonyms

Synonyms Comment Organism
PpoA
-
Aspergillus nidulans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
24
-
assay at Aspergillus nidulans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5
-
palmitoleate pH 7.2, 24°C Aspergillus nidulans
4.95
-
oleate pH 7.2, 24°C Aspergillus nidulans
5.9
-
(11Z)-icos-11-enoic acid pH 7.2, 24°C Aspergillus nidulans
6.3
-
linoleate pH 7.2, 24°C Aspergillus nidulans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.2
-
assay at Aspergillus nidulans

Cofactor

Cofactor Comment Organism Structure
cytochrome P450 PpoA contains a high spin ferriheme. PpoA uses different heme domains to catalyze two separate reactions. Within the N-terminal heme peroxidase domain, linoleic acid is oxidized to (8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate by abstracting a H-atom from C-8 of the fatty acid, yielding a carbon-centered radical that reacts with molecular dioxygen. In the second reaction step, (8R,9Z,12Z)-8-hydroperoxy-9,12-octadecadienoate is isomerized within the C-terminal P450 heme thiolate domain to (5S,8R,9Z,12Z)-5,8-dihydroxy-9,12-octadecadienoate Aspergillus nidulans
heme PpoA contains a high spin ferriheme. PpoA uses different heme domains to catalyze two separate reactions. Within the N-terminal heme peroxidase domain, linoleic acid is oxidized to (8R)-hydroperoxyoctadecadienoic acid by abstracting a H-atom from C-8 of the fatty acid, yielding a carbon-centered radical that reacts with molecular dioxygen. In the second reaction step, 8-hydroperoxyoctadecadienoic acid is isomerized within the C-terminal P450 heme thiolate domain to 5,8-dihydroxyoctadecadienoic acid Aspergillus nidulans

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
261
-
(11Z)-icos-11-enoic acid pH 7.2, 24°C Aspergillus nidulans
344
-
linoleate pH 7.2, 24°C Aspergillus nidulans
700
-
palmitoleate pH 7.2, 24°C Aspergillus nidulans
738
-
oleate pH 7.2, 24°C Aspergillus nidulans