Information on EC 1.14.13.2 - 4-hydroxybenzoate 3-monooxygenase

New: Word Map on EC 1.14.13.2
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Search Reference ID:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
1.14.13.2
-
RECOMMENDED NAME
GeneOntology No.
4-hydroxybenzoate 3-monooxygenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
4-hydroxybenzoate + NADPH + H+ + O2 = protocatechuate + NADP+ + H2O
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
4-chlorobenzoate degradation
-
-
4-hydroxymandelate degradation
4-methylphenol degradation to protocatechuate
-
-
Benzoate degradation
-
-
bisphenol A degradation
-
-
Metabolic pathways
-
-
Microbial metabolism in diverse environments
-
-
SYSTEMATIC NAME
IUBMB Comments
4-hydroxybenzoate,NADPH:oxygen oxidoreductase (3-hydroxylating)
A flavoprotein (FAD). Most enzymes from Pseudomonas are highly specific for NADPH (cf. EC 1.14.13.33 4-hydroxybenzoate 3-monooxygenase [NAD(P)H]).
CAS REGISTRY NUMBER
COMMENTARY hide
9059-23-8
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain ADPU54
-
-
Manually annotated by BRENDA team
strain ADPU54
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain PPH
-
-
Manually annotated by BRENDA team
strain PPH
-
-
Manually annotated by BRENDA team
Burkholderia xenovorans
-
-
-
Manually annotated by BRENDA team
strain HS-2
-
-
Manually annotated by BRENDA team
strain HS-2
-
-
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
Comamonas testosteroni Kh 122-3S
Kh 122-3S
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
enzyme is expressed in mutant strain MAO4, but not in wild-type strain
-
-
Manually annotated by BRENDA team
strain GU2
-
-
Manually annotated by BRENDA team
strain GU2
-
-
Manually annotated by BRENDA team
KR1
-
-
Manually annotated by BRENDA team
KR1
-
-
Manually annotated by BRENDA team
A 3.12
-
-
Manually annotated by BRENDA team
M-6
-
-
Manually annotated by BRENDA team
strain WCS358
UniProt
Manually annotated by BRENDA team
CBS3
-
-
Manually annotated by BRENDA team
strain PPD
-
-
Manually annotated by BRENDA team
557, 420, and 1G
-
-
Manually annotated by BRENDA team
135
-
-
Manually annotated by BRENDA team
135
-
-
Manually annotated by BRENDA team
172
-
-
Manually annotated by BRENDA team
400
-
-
Manually annotated by BRENDA team
400
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
Burkholderia xenovorans
-
the pobA gene encoding the 4-hydroxybenzoate 3-monooxygenase is expressed during growth on hydroxybenzoic acids and glucose
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2,3-dihydroxybenzoate + NADPH + O2
? + NADP+ + H2O
show the reaction diagram
2,4-dihydroxybenzoate + NADPH + O2
2,3,4-trihydroxybenzoate + 2,4,5-trihydroxybenzoate + NADP+ + H2O
show the reaction diagram
2,5-dihydroxybenzoate + NADPH + O2
? + NADP+ + H2O
show the reaction diagram
2-chloro-4-hydroxybenzoate + NADH + O2
?
show the reaction diagram
2-fluoro-4-hydroxybenzoate + NADH + O2
?
show the reaction diagram
3,4-dihydroxybenzoate + NADPH + O2
? + NADP+ + H2O
show the reaction diagram
3,5-dihydroxybenzoate + NADPH + O2
? + NADP+ + H2O
show the reaction diagram
3-bromo-4-hydroxybenzoate + NADPH + O2
?
show the reaction diagram
-
3.2% of the activity with 4-hydroxybenzoate
-
-
?
3-Chloro-4-hydroxybenzoate + NADPH + O2
?
show the reaction diagram
-
6.5% of the activity with 4-hydroxybenzoate
-
-
?
3-chlorophenol + NADPH + O2
? + NADP+ + H2O
show the reaction diagram
-
-
-
?
3-Fluoro-4-hydroxybenzoate + NADPH + O2
?
show the reaction diagram
-
about 1% of the activity with 4-hydroxybenzoate
-
-
?
3-hydroxyanthranilate + NADPH + O2
? + NADP+ + H2O
show the reaction diagram
-
-
-
?
4-aminobenzoate + NADPH + O2
?
show the reaction diagram
-
about 1% of the activity with 4-hydroxybenzoate
-
-
?
4-chlorophenol + NADPH + O2
? + NADP+ + H2O
show the reaction diagram
-
-
-
?
4-chlororesorcinol + NADPH + O2
? + NADP+ + H2O
show the reaction diagram
-
-
-
?
4-hydroxybenzoate + NADH + O2
protocatechuate + NAD+ + H2O
show the reaction diagram
4-hydroxybenzoate + NADPH + ferricyanide
protocatechuate + NADP+ + ferrocyanide
show the reaction diagram
-
-
-
?
4-hydroxybenzoate + NADPH + H+ + O2
protocatechuate + NADP+ + H2O
show the reaction diagram
4-hydroxybenzoate + NADPH + O2
?
show the reaction diagram
4-hydroxybenzoate + NADPH + O2
protocatechuate + NADP+ + H2O
show the reaction diagram
4-mercaptobenzoate + NADPH + O2
4,4'-dithiobisbenzoate + ?
show the reaction diagram
4-nitrophenol + NADPH + O2
? + NADP+ + H2O
show the reaction diagram
-
-
-
?
4-toluate + NADPH + O2
?
show the reaction diagram
-
0.29% of the activity with 4-hydroxybenzoate
-
-
?
benzene sulfonate + NADPH + O2
?
show the reaction diagram
-
0.34% of the activity with 4-hydroxybenzoate
-
-
?
hydroquinone + NADP+ + O2
? + NADP+ + H2O
show the reaction diagram
-
-
-
?
m-hydroxybenzoate + NADPH + O2
protocatechuate + NADP+ + H2O
show the reaction diagram
-
-
-
?
p-hydroxybenzoate + NADPH + O2
? + NADP+ + H2O
show the reaction diagram
is transformed to a lesser extent than m-hydroxybenzoate
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
4-hydroxybenzoate + NADPH + H+ + O2
protocatechuate + NADP+ + H2O
show the reaction diagram
4-hydroxybenzoate + NADPH + O2
?
show the reaction diagram
4-hydroxybenzoate + NADPH + O2
protocatechuate + NADP+ + H2O
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-deaza-FAD
-
when 1-deaza-FAD is used as cofactor, the enzyme carries out each step in catalysis except the transfer of oxygen to 4-hydroxybenzoate
6-Hydroxy-FAD
-
when 6-hydroxy-FAD is used as cofactor, the enzyme has a lower turnover rate than the native enzyme
arabinoflavin adenine dinucleotide
-
like native enzyme the arabinoflavin adenine dinucleotide containing 4-hydroxybenzoate hydroxylase preferentially binds the phenolate form of the substrate. The oxidative part of the catalytic cycle of a FAD-containing 4-hydroxybenzoate hydroxylase differs from the native enzyme. Partial uncoupling of hydroxylation results in the formation of about 0.3 mol of 3,4-dihydroxybenzoate and 0.7 mol of H2O2 per mol of NADPH oxidized
additional information
-
helix H2 is involved in determining the coenzyme specificity
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
metal ions (at 1 mM) Co2+, Ni2+, Mg2+, and Hg2+ shows little influence on hydroxylase activity
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(-)-epigallocatechin-3-O-gallate
-
non-competitive, binds to the enzyme in the proximity of the FAD binding site via formation of three hydrogen bonds
2,4-Dihydroxybenzoate
-
competitive with 4-hydroxybenzoate
3,4-dihydroxybenzoate
3-Chloro-4-hydroxybenzoate
-
mixed-type
3-hydroxybenzoate
4-Aminobenzoate
4-Aminosalicylate
-
competitive
4-Fluorobenzoate
4-hydroxy-3-nitrobenzoic acid
-
2 mM, 23% inhibition
4-hydroxybenzaldehyde
4-hydroxybenzoate
4-hydroxybutyrate
-
92 mM, pH 8, very slight inhibition
4-hydroxycinnamate
-
-
4-hydroxyphenylacetic acid
-
2 mM, 16% inhibition
4-nitrophenol
-
2 mM, 39% inhibition
6-Aminonicotinate
-
competitive
6-Hydroxynicotinate
acetate
-
92 mM, pH 8, very slight inhibition
Benzoate
citrate
-
92 mM, pH 8, very slight inhibition
coumaric acid
-
2 mM, 30% inhibition
Cu2+
-
1 mM, moderate inhibition
diethyl dicarbonate
Fe3+
-
1 mM, moderate inhibition
formate
-
92 mM, pH 8, very slight inhibition
fumarate
-
92 mM, pH 8, very slight inhibition
Hg2+
-
0.1 mM HgCl2, complete inhibition
iodoacetamide
-
reversed by dithiothreitol
Maleate
-
92 mM, pH 8, very slight inhibition
Mn2+
-
1 mM, moderate inhibition
n-dodecyl gallate
-
non-competitive
N-iodosuccinimide
-
reversed by dithiothreitol
NEM
-
0.1 mM, 59% inhibition
o-Iodosobenzoate
-
reversed by dithiothreitol
p-hydroxy-3-iodomethylbenzoate
-
1 mM, irreversible crosslinking to the substrate binding site
Phenylglyoxal
-
pseudo-first order kinetics, incorporation into the substrate-binding site
phosphate
Propionate
-
92 mM, pH 8, very slight inhibition
protocatechuate
-
above 1 mM
salicylate
Tartrate
-
92 mM, pH 8, very slight inhibition
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,4-Dihydroxybenzoate
-
increases the rate of NADP oxidation by 4-hydroxybenzoate, no hydroxylation during subsequent reoxidation by O2
3,4-dihydroxybenzoate
-
increases the rate of NADP oxidation by 4-hydroxybenzoate, no hydroxylation during subsequent reoxidation by O2
5-Hydroxypicolinate
-
stimulates rapid oxidation of NADPH
Benzoate
-
increases the rate of NADP oxidation by 4-hydroxybenzoate, no hydroxylation during subsequent reoxidation by O2
dithiothreitol
-
4 mM, slightly increases activity
EDTA
-
5 mM or 20 mM, slightly increases activity
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.014 - 0.25
2,4-Dihydroxybenzoate
0.18
3-Chloro-4-hydroxybenzoate
-
-
0.0006 - 0.11
4-hydroxybenzoate
0.0122 - 0.18
NADH
0.027
NADP+
-
reaction with 2,4-dihydroxybenzoate
0.0226 - 0.32
NADPH
0.037 - 0.05
O2
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.36 - 63
4-hydroxybenzoate
6.8 - 16.9
NADH
6.6 - 63
NADPH
0.2 - 7.5
p-hydroxybenzoate
additional information
additional information
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.014
(-)-epigallocatechin-3-O-gallate
-
-
0.31
2,4-Dihydroxybenzoate
-
-
0.55
3,4-dihydroxybenzoate
-
-
0.05
4-Aminobenzoate
-
-
5.9 - 50
Cl-
0.0181
n-dodecyl gallate
-
-
4.3
phosphate
-
4C, pH 6.8
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 7.3
-
reaction with NADH
7.2
-
strain 400 and 557
7.5 - 7.8
-
reaction with NADPH
8 - 8.1
-
HEPES or Tris-SO42- buffer
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
44000
estimated from amino acid sequence
65000
-
gel filtration
75000
-
gel filtration
76000 - 82000
-
gel filtration
78000
-
gel filtration
83000 - 90000
-
equilibrium sedimentation
83600
-
low speed sedimentation without reaching equilibrium
85000
-
gel filtration
89000
-
gel filtration
90000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 71000, SDS-PAGE
monomer
-
the monomeric form of PHBH, which is not achieved under conventional conditions, is isolated by entrapment in reverse micelles and by addition of DMSO. The PHBH monomer is catalytically more efficient than the PHBH dimer
tetramer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging-drop vapour diffusion method in the presence of NaH2PO4 and K2HPO4 as precipitants. X-ray diffraction data are collected to a maximum resolution of 2.5 A on a synchrotron beamline. The crystal belongs to the hexagonal space group P6(3)22, with unit-cell parameters a = b = 94.72, c = 359.68 A, gamma = 120. The asymmetric unit contains two molecules
-
crystal structure of mutant enzyme Y201F, Y385F, and N300D
-
hanging drop vapor diffusion, hanging drops containing 4 mg/ml protein, 100 mM potassium phosphate, pH 7.0, 0.05 mM glutathione, 30 mM sodium sulfite, 0.02 mM FAD, 450 mM ammonium sulfate are equilibrated at 30C for 7-10 days against a well solution of similar composition, but containing 900 mM ammonium sulfate, crystals of R220Q PHBH diffract to approx. 2.0 A
-
crystal of the enzyme complexed with 4-hydroxybenzoate are obtained using the hanging-drop method
-
crystal structure of wild-type p-hydroxybenzoate hydroxylase complexed with 4-aminobenzoate, 2,4-dihydroxybenzoate, and 2-hydroxy-4-aminobenzoate and of the Tyr222Ala mutant complexed with 2-hydroxy-4-aminobenzoate
-
crystallization of mutant enzymes H162R and R269T by hanging drop vapour diffusion method
-
crystals of a arabinoflavin adenine dinucleotide -containing 4-hydroxybenzoate hydroxylase in complex with 4-hydroxybenzoate are obtained using the hanging drop method
-
enzyme bound in a crystal is able to convert 4-hydroxybenzoate
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 6.5
-
optimal stability
390027
6.4 - 8
-
at 0C or at 25C, 50 h, stable
390018
7 - 7.5
-
optimal stability
390036
9 - 9.5
-
optimal stability
390036
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
21
-
room temperature, without glycerol and EDTA, complete inactivation after 20 h
52
-
pH 7.5, 20 mM potassium phosphate buffer, 20 min, stable
65
-
1 mM 4-hydroxybenzoate, 95% loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
enzyme rapidly loses activity in dilute solutions, below 2 mg/ml, without stabilizer
extremely unstable, undergoing rapid inactivation unless protected by substrates and other stabilizing agents
-
stable to freezing and thawing
-
very stable, even in absence of stabilizing agents
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-70C, 5 mM potassium phosphate, pH 7.5, 5% glycerol, stable for several months
-
0C-4C, as a precipitate under a solution of 50 mM potassium phosphate and 0.5 mM EDTA, pH 6.5-7.0, with 70% saturated ammonium sulfate, idefinitely stable
-
4C, 24 h, 40% loss of activity without stabilizer, 10% loss of activity in presence of 4-hydroxybenzoate, FAD and EDTA
-
4C, 50 mM phosphate buffer, 50 mM Tris-HCl, pH 8.0, 10% glycerol, 10 mM EDTA, loss of activity after 72 h
-
4C, as ammonium sulfate paste, indefinitely stable
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
microheterogeneity of the highly purified enzyme. Different form of enzyme molecules are due to the partial oxidation of Cys116 in the sequence of the enzyme
-
recombinant enzyme
-
recombinant PHBH
-
recombinat enzyme, cloned in Escherichia coli
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
expression in Escherichia coli
-
expression in Escherichia coli, mutant enzyme S212A
-
expression of mutant enzymes Y201F and Y385F in Escherichia coli
-
expression of wild-type and E49Q mutant PHPH in Escherichia coli
-
mobA gene from pSEB2 cloned into pZR80. Random mutations introduced into the mobA gene and cloned into pCR2.1-TOPO and transformed into Escherichia coli Top10
mutant enzymes K297M, N300D and Y385F
-
mutant enzymes R42S and R42K expressed in transformed Escherichia coli TG2 cells
-
overexpression in Escherichia coli
-
plasmid mutagenesis for high-level expression of 4-hydroxybenzoate hydroxylase
-
When expressed in Escherichia coli, p-hydroxybenzoate hydroxylase transforms p-hydroxybenzoate into protocatechuate
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
4-hydroxybenzoate 3-hydroxylase-encoding pobA transcripts are nearly absent in presence of benzoate and benzoate drastically decreases the transcription of this gene. Repression is mediated by pobR, the transcriptional activator of pobA gene
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A16T/S394P/D416A
low ability to hydroxylate 3-aminophenol
A400G
transforms 3-aminophenol with efficiency almost like mutant A400G/K429R
A400G/K429R
among mutants, highest enzymatic activity to hydroxylate 3-aminophenol
H135P
alters the enzyme's substrate specificity; low ability to hydroxylate 3-aminophenol
H135P/I217L/Y304H
low ability to hydroxylate 3-aminophenol
K326I
lacks the ability to transform phenol to catechol as the wild-type
K429R
can not transform 3-aminophenol at all
N102T/I259S/V399M
low ability to hydroxylate 3-aminophenol
N227H
is not able to transform 3-aminophenol
N227H/D416A
almost has the same transformation efficiency as mutant N227H/Q292R/D416A
N227H/Q292R/D416A
among mutants, highest enzymatic activity to hydroxylate 3-aminophenol
Q292R
is not able to transform 3-aminophenol
R152L/F364V
low ability to hydroxylate 3-aminophenol
V257A
mutation enables the mutant to transform phenol to catechol, also has enhanced ability to transform resorcinol, hydroquinone, p-hydroxybenzoate, 2,5-dihydroxybenzoate, 3,4-dihydroxybenzoate, 3-chlorophenol, 4-chlorophenol, 4-chlororesorcinol, and 4-nitrophenol, thus broadens the substrate range. Is not capable of hydroxylating benzoate, o-hydroxybenzoate (salicylate), 2,4-dihydroxybenzoate, 2,6-dihydroxybenzoate, 2-chlorophenol, 3-aminophenol, 4-methoxybenzoate, 3-toluate, o-cresol, m-cresol, or p-cresol as the wild-type
A400G
-
transforms 3-aminophenol with efficiency almost like mutant A400G/K429R
-
H135P
-
alters the enzyme's substrate specificity; low ability to hydroxylate 3-aminophenol
-
K326I
-
lacks the ability to transform phenol to catechol as the wild-type
-
V257A
-
mutation enables the mutant to transform phenol to catechol, also has enhanced ability to transform resorcinol, hydroquinone, p-hydroxybenzoate, 2,5-dihydroxybenzoate, 3,4-dihydroxybenzoate, 3-chlorophenol, 4-chlorophenol, 4-chlororesorcinol, and 4-nitrophenol, thus broadens the substrate range. Is not capable of hydroxylating benzoate, o-hydroxybenzoate (salicylate), 2,4-dihydroxybenzoate, 2,6-dihydroxybenzoate, 2-chlorophenol, 3-aminophenol, 4-methoxybenzoate, 3-toluate, o-cresol, m-cresol, or p-cresol as the wild-type
-
D38A
-
kcat/KM for 4-hydroxybenzoate is 16.8fold higher than wild-type value
D38Y
-
kcat/KM for 4-hydroxybenzoate is 11.8fold higher than wild-type value
D38Y/T42R
-
kcat/KM for 4-hydroxybenzoate is 32fold higher than wild-type value
T42R
-
kcat/KM for 4-hydroxybenzoate is 7.2fold higher than wild-type value
P293S
-
mutation decreases the stability of the folded mutant protein compared to the wild-type PHBH
R220Q
-
1% of wild-type activity, lower affinity to 4-hydroxybenzoate than wild-type
S212A
-
the turnover of the substrate 2,4-dihydroxybenzoate is 1.5-fold faster than the rate observed with the wild-type
H162D
-
no reliable turnover rate due to impaired NADPH binding
H162K
-
less efficient than wild-type enzyme due to a clear increase in the apparent Km-value for NADPH
H162N
-
no reliable turnover rate due to impaired NADPH binding
H162R
-
rather efficient enzyme with similar catalytic properties as wild-type enzyme
H162S
-
no reliable turnover rate due to impaired NADPH binding
H162T
-
no reliable turnover rate due to impaired NADPH binding
H162Y
-
rather efficient enzyme with similar catalytic properties as wild-type enzyme
R269D
-
no reliable turnover rate due to impaired NADPH binding
R269K
-
rather efficient enzyme with similar catalytic properties as wild-type enzyme
R269N
-
no reliable turnover rate due to impaired NADPH binding
R269S
-
less efficient than wild-type enzyme due to a clear increase in the apparent Km-value for NADPH
R269T
-
no reliable turnover rate due to impaired NADPH binding
R269Y
-
no reliable turnover rate due to impaired NADPH binding
R42K
-
low activity results from impaired binding of NADPH
R42S
-
low activity results from impaired binding of NADPH
Y222A
-
mutation makes the lifetime distribution of FAD in the enzyme simpler by removing the ultrafast 10-15 ps lifetime component
Y222V
-
mutation makes the lifetime distribution of FAD in the enzyme simpler by removing the ultrafast 10-15 ps lifetime component
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
-
a bienzyme-based Clark electrode is developed for the interference-free determination of L-glutamate. This sensor is based on the specific dehydrogenation by L-glutamate dehydrogenase (EC 1.4.1.3) in combination with p-hydroxybenzoate hydroxylase (EC 1.14.13.2). The enzymes are entrapped by a poly(carbamoyl) sulfonate hydrogel on a Teflon membrane
degradation
Show AA Sequence (1679 entries)
Please use the Sequence Search for a certain query.