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

BRENDA support

BRENDA Home
show all | hide all No of entries

Information on EC 1.14.99.15 - 4-methoxybenzoate monooxygenase (O-demethylating)

for references in articles please use BRENDA:EC1.14.99.15
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
IUBMB Comments
The bacterial enzyme consists of a ferredoxin-type protein and an iron-sulfur flavoprotein (FMN). Also acts on 4-ethoxybenzoate, N-methyl-4-aminobenzoate and toluate. The fungal enzyme acts best on veratrate.
Specify your search results
Select one or more organisms in this record: ?
This record set is specific for:
UNIPROT: Q2IU02
Show additional data
Do not include text mining results
Include (text mining) results
Include results (AMENDA + additional results, but less precise)
Word Map
hide
The enzyme appears in viruses and cellular organisms
Synonyms
cyp199a2, cyp199a4, 4-methoxybenzoate monooxygenase, 4-methoxybenzoate o-demethylase, 4-methoxybenzoate monooxygenase (o-demethylating), more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4-methoxybenzoate 4-monooxygenase (O-demethylating)
-
-
-
-
4-methoxybenzoate O-demethylase
-
-
-
-
oxygenase, 4-methoxybenzoate 4-mono- (O-demethylating)
-
-
-
-
p-anisic O-demethylase
-
-
-
-
piperonylate-4-O-demethylase
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
redox reaction
-
-
-
-
oxidation
-
-
-
-
reduction
-
-
-
-
O-demethylation
-
-
-
-
S-demethylation
-
-
-
-
N-demethylation
-
-
-
-
dealkylation
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
4-methoxybenzoate,hydrogen-donor:oxygen oxidoreductase (O-demethylating)
The bacterial enzyme consists of a ferredoxin-type protein and an iron-sulfur flavoprotein (FMN). Also acts on 4-ethoxybenzoate, N-methyl-4-aminobenzoate and toluate. The fungal enzyme acts best on veratrate.
CAS REGISTRY NUMBER
COMMENTARY hide
37256-78-3
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4-methoxyacetophenone + AH2 + O2
4-hydroxyacetophenone + formaldehyde + A + H2O
show the reaction diagram
low activity
-
-
?
4-methoxybenzaldehyde + AH2 + O2
4-hydroxybenzaldehyde + formaldehyde + A + H2O
show the reaction diagram
low activity
-
-
?
4-methoxybenzamide + AH2 + O2
4-hydroxybenzamide + formaldehyde + A + H2O
show the reaction diagram
low activity
-
-
?
4-methoxybenzoate + AH2 + O2
4-hydroxybenzoate + formaldehyde + A + H2O
show the reaction diagram
4-methoxyphenylacetate + AH2 + O2
4-hydroxyphenylacetate + formaldehyde + A + H2O
show the reaction diagram
low activity
-
-
?
4-methoxyphenylboronic acid + AH2 + O2
4-hydroxyphenylboronic acid + formaldehyde + A + H2O
show the reaction diagram
low activity
-
-
?
4-methylbenzoate + AH2 + O2
?
show the reaction diagram
low activity
-
-
?
4-methylphenylacetate + AH2 + O2
?
show the reaction diagram
low activity
-
-
?
veratrate + AH2 + O2
?
show the reaction diagram
-
-
-
?
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
4-methoxybenzoate + AH2 + O2
4-hydroxybenzoate + formaldehyde + A + H2O
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Cl-
addition of chloride to the phosphate buffered samples weakens substrate binding, chloride binding site structure, overview
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
the enzyme belongs to the superfamily of heme-dependent cytochrome P450 monooxygenases
additional information
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
Q2IU02_RHOP2
Rhodopseudomonas palustris (strain HaA2)
410
0
44538
TrEMBL
-
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
enzyme CYP199A4 free and bound to substrate 4-methoxybenzoate, hanging drop vapour diffusion method, for the free enzyme: mixing of 0.001 ml of protein solution containing 50 mg/ml protein in 20 mM HEPES, pH 7.4, 150 mM KCl, 1 mM DTT, with 0.001 ml of reservoir solution containing 0.1 M Bis-Tris, pH 5.5, 1.45-1.5 M ammonium sulfate, and 0.1 M sodium chloride, and equilibration against 0.2 ml of reservoir solution, 20°C, 1 week, for the substrate-bound enzyme: mixing of 0.001 ml of protein solution containing 40 mg/ml protein in 20 mM HEPES, pH 7.4, 150 mM KCl, and 10 mM 2-mercaptoethanol and saturated with 4-methoxybenzoate, with 0.001 ml of reservoir solution containing 0.1 M Bis-Tris, pH 5.5, 1.45 M ammonium sulfate, and 0.1 M sodium chloride, and equilibration against 0.2 ml of reservoir solution, 20°C, 2 weeks, X-ray diffraction structure determination and analysis at 2.6 A and 2.0 A resolution, respectively
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
F185I
site-directed mutagenesis, the mutation reduces the spin state shift from low- to high-spin on the addition of 4-methoxybenzoate by about 25% compared to the wild-type enzyme, the mutant shows reduced NADH consumption and product formation
F185V
site-directed mutagenesis, the mutation reduces the spin state shift from low- to high-spin on the addition of 4-methoxybenzoate by about 35% compared to the wild-type enzyme, the mutant shows reduced NADH consumption and product formation
R243T
site-directed mutagenesis, the mutation reduces the spin state shift from low- to high-spin on the addition of 4-methoxybenzoate by about 25% compared to the wild-type enzyme, the mutant shows reduced NADH consumption and product formation
R92E
site-directed mutagenesis, the spin state shift is similar to the wild-type enzyme, but the mutant shows 3fold higher KD for the substrate, NADH consumption is reduced 9fold compared to the wild-type enzyme
S95V
site-directed mutagenesis, the mutation abolishes the spin state shift from low- to high-spin on the addition of 4-methoxybenzoate and results in a 99% drop in the NADH consumption rate comared to the wild-type enzyme
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by two different steps of anion exchange chromatography, followed by gel filtration
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Bell, S.; Yang, W.; Tan, A.; Zhou, R.; Johnson, E.; Zhang, A.; Zhou, W.; Rao, Z.; Wong, L.
The crystal structures of 4-methoxybenzoate bound CYP199A2 and CYP199A4: Structural changes on substrate binding and the identification of an anion binding site
Dalton Trans.
41
8703-8714
2012
Rhodopseudomonas palustris (Q2IU02), Rhodopseudomonas palustris (Q6N8N2), Rhodopseudomonas palustris, Rhodopseudomonas palustris CGA009 (Q6N8N2), Rhodopseudomonas palustris HaA2 (Q2IU02)
Manually annotated by BRENDA team
Coleman, T.; Chao, R.R.; De Voss, J.J.; Bell, S.G.
The importance of the benzoic acid carboxylate moiety for substrate recognition by CYP199A4 from Rhodopseudomonas palustris HaA2
Biochim. Biophys. Acta
1864
667-675
2016
Rhodopseudomonas palustris (Q2IU02), Rhodopseudomonas palustris HaA2 (Q2IU02), Rhodopseudomonas palustris HaA2
Manually annotated by BRENDA team