Information on EC 3.5.1.32 - hippurate hydrolase

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

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EC NUMBER
COMMENTARY hide
3.5.1.32
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RECOMMENDED NAME
GeneOntology No.
hippurate hydrolase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
hippurate + H2O = benzoate + glycine
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
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-
-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Phenylalanine metabolism
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SYSTEMATIC NAME
IUBMB Comments
N-benzoylamino-acid amidohydrolase
Acts on various N-benzoylamino acids.
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CAS REGISTRY NUMBER
COMMENTARY hide
37278-43-6
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
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Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
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-
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
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-
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Manually annotated by BRENDA team
strain ATCC 43431, gene hipO
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Manually annotated by BRENDA team
no activity in Bacillus lentus
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-
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Manually annotated by BRENDA team
no activity in Bacillus macquariensis
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-
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Manually annotated by BRENDA team
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4-acetyl-N-benzoyl-L-alanine + H2O
4-acetylbenzoic acid + L-alanine
show the reaction diagram
-
low activity
-
?
4-acetyl-N-benzoylglycine + H2O
4-acetylbenzoic acid + glycine
show the reaction diagram
-
-
-
?
4-aminobenzoyl-glutamate + H2O
4-aminobenzoate + glutamate
show the reaction diagram
-
-
-
-
?
4-chloro-N-benzoyl-L-alanine + H2O
4-chlorobenzoic acid + L-alanine
show the reaction diagram
-
-
-
?
4-chloro-N-benzoylglycine + H2O
4-chlorobenzoic acid + glycine
show the reaction diagram
-
-
-
?
4-hydroxy-N-benzoyl-L-alanine
4-hydroxybenzoic acid + L-alanine
show the reaction diagram
-
low activity
-
?
4-hydroxy-N-benzoylglycine + H2O
4-hydroxybenzoic acid + glycine
show the reaction diagram
-
-
-
?
4-hydroxy-N-benzoylglycine-L-histidyl-L-leucine + H2O
4-hydroxybenzoic acid + L-histidyl-L-leucine
show the reaction diagram
-
-
-
?
4-nitro-N-benzoyl-L-alanine + H2O
4-nitrobenzoic acid + L-alanine
show the reaction diagram
-
-
-
?
4-nitro-N-benzoylglycine + H2O
4-nitrobenzoic acid + glycine
show the reaction diagram
-
best substrate
-
?
4-nitro-N-benzoylglycylglycylglycine + H2O
4-nitrobenzoic acid + glycylglycylglycine
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-glycyl-L-phenylalanine + H2O
benzoic acid + ?
show the reaction diagram
-
-
-
?
glycyl-L-leucine + H2O
glycine + L-leucine
show the reaction diagram
-
-
-
?
glycylglycylglycine + H2O
glycine + glycylglycine
show the reaction diagram
-
-
-
?
hippurate + H2O
benzoate + glycine
show the reaction diagram
itoic acid + H2O
2,3-dihydroxybenzoic acid + glycine
show the reaction diagram
-
low specificity
-
?
N-acetyl-DL-alanine + H2O
acetic acid + DL-alanine
show the reaction diagram
-
-
-
?
N-acetyl-DL-asparagine + H2O
acetic acid + DL-asparagine
show the reaction diagram
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low activity
-
?
N-acetyl-DL-methionine + H2O
acetic acid + DL-methionine
show the reaction diagram
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low activity
-
?
N-acetyl-DL-phenylalanine + H2O
acetic acid + DL-phenylalanine
show the reaction diagram
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low activity
-
?
N-acetyl-DL-tryptophan + H2O
acetic acid + DL-tryptophan
show the reaction diagram
-
-
-
?
N-acetyl-DL-valine + H2O
acetic acid + DL-valine
show the reaction diagram
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very low activity
-
?
N-acetyl-glycine + H2O
acetic acid + glycine
show the reaction diagram
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low activity
-
?
N-acetyl-L-cysteine + H2O
acetic acid + L-cysteine
show the reaction diagram
-
-
-
?
N-acetyl-L-glutamine + H2O
acetic acid + L-glutamine
show the reaction diagram
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low activity
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?
N-acetylalanine + H2O
acetate + alanine
show the reaction diagram
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-
-
-
?
N-acetylglycine + H2O
acetate + glycine
show the reaction diagram
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-
-
-
?
N-benzoyl-(S-benzoyl)-DL-cysteine + H2O
benzoic acid + S-benzoyl-DL-cysteine
show the reaction diagram
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low activity
-
?
N-benzoyl-DL-alanine + H2O
benzoic acid + DL-alanine
show the reaction diagram
-
-
-
?
N-benzoyl-DL-alloisoleucine + H2O
benzoic acid + DL-alloisoleucine
show the reaction diagram
-
-
-
?
N-benzoyl-DL-arginine + H2O
benzoic acid + DL-arginine
show the reaction diagram
-
-
-
?
N-benzoyl-DL-asparagine + H2O
benzoic acid + DL-asparagine
show the reaction diagram
-
-
-
?
N-benzoyl-DL-glutamine + H2O
benzoic acid + DL-glutamine
show the reaction diagram
-
-
-
?
N-benzoyl-DL-histidine + H2O
benzoic acid + DL-histidine
show the reaction diagram
-
-
-
?
N-benzoyl-DL-leucine + H2O
benzoic acid + DL-leucine
show the reaction diagram
-
-
-
?
N-benzoyl-DL-methionine + H2O
benzoic acid + DL-methionine
show the reaction diagram
N-benzoyl-DL-phenylalanine + H2O
benzoic acid + DL-phenylalanine
show the reaction diagram
-
-
-
?
N-benzoyl-DL-serine + H2O
benzoic acid + DL-serine
show the reaction diagram
-
-
-
?
N-benzoyl-DL-threonine + H2O
benzoic acid + DL-threonine
show the reaction diagram
-
-
-
?
N-benzoyl-DL-tryptophan + H2O
benzoic acid + DL-tryptophan
show the reaction diagram
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low activity
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?
N-benzoyl-DL-tyrosine + H2O
benzoic acid + DL-tyrosine
show the reaction diagram
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-
-
?
N-benzoyl-DL-valine + H2O
benzoic acid + DL-valine
show the reaction diagram
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low activity
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?
N-benzoyl-L-alanine + H2O
benzoic acid + L-alanine
show the reaction diagram
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-
-
?
N-benzoyl-L-aminobutyric acid + H2O
benzoic acid + L-aminobutyric acid
show the reaction diagram
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low activity
-
?
N-benzoyl-L-valine + H2O
benzoic acid + L-valine
show the reaction diagram
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poor substrate
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?
N-benzoylalanine + H2O
benzoate + alanine
show the reaction diagram
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-
-
-
?
N-benzoylglutamate + H2O
benzoate + glutamate
show the reaction diagram
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-
-
-
?
N-benzoylglycine + H2O
benzoic acid + glycine
show the reaction diagram
N-benzoylglycylglycylglycine + H2O
benzoic acid + glycylglycylglycine
show the reaction diagram
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-
-
?
N-benzoylmethionine + H2O
benzoate + methionine
show the reaction diagram
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-
-
-
?
N-benzoylphenylalanine + H2O
benzoate + phenylalanine
show the reaction diagram
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-
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?
N-benzoylvaline + H2O
benzoate + valine
show the reaction diagram
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?
additional information
?
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substrate specificity, the enzyme also shows metallocarboxypeptidase activity with carboxypeptidase substrates in vitro, displaying its highest activity against N-benzoyl-linked small aliphatic acids, overview, the enzyme does not hydrolyze typical substrates for aminopeptidases, lysine-p-nitroanilide, leucine-p-nitroanilide, or endopeptidases, N-benzoylarginine-p-nitroanilide, succinylphenylalanine-4-nitroanilide
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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
hippurate + H2O
benzoate + glycine
show the reaction diagram
-
-
-
-
?
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ba2+
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10% activation at 10 mM
K+
-
20% activation at 10 mM
Mg2+
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20% activation at 10 mM
Zn2+
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1.98 molecules of zinc per enzyme subunit molecule, metalloenzyme
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
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99% inhibition at 0.5 mM
2,2'-dipyridyl
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80% inhibition at 1 mM
2-mercaptoethanol
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20% inhibition at 2 mM
8-hydroxyquinoline
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60% inhibition at 3.3 mM
Ag+
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98% inhibition at 2 mM
Ba2+
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slight inhibition at 1 mM
Cd2+
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65% inhibition at 2 mM
EDTA
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40% inhibition at 10 mM
Fe3+
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42% inhibition at 1 mM
iodoacetic acid
Mn2+
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60% inhibition at 1 mM
o-phenanthroline
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complete inhibition at 1 mM
p-chloromercuribenzoic acid
Zn2+
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35% inhibition at 2 mM
additional information
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no significant inhibition by E64, PMSF, and pepstatin at 0.5 mM
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
EDTA
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23.9% activation at 10 mM
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.1
4-acetyl-N-benzoyl-L-alanine
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-
0.54
4-acetyl-N-benzoylglycine
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-
1.769
4-aminobenzoyl-glutamate
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pH 7.5, 37°C, purified recombinant wild-type enzyme
0.71
4-chloro-N-benzoyl-L-alanine
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-
0.37
4-chloro-N-benzoylglycine
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-
0.95
4-hydroxy-N-benzoyl-L-alanine
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-
1.25
4-nitro-N-benzoyl-L-alanine
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1.2
4-nitro-N-benzoylglycine
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0.0006 - 2.778
hippurate
5.45
N-acetyl-L-alanine
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pH 7.5, 37°C, purified recombinant wild-type enzyme
3.377
N-acetylglycine
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pH 7.5, 37°C, purified recombinant wild-type enzyme
0.87
N-benzoyl-L-alanine
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-
0.87
N-benzoyl-L-aminobutyric acid
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-
0.169
N-benzoylalanine
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pH 7.5, 37°C, purified recombinant wild-type enzyme
0.159
N-benzoylglutamate
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pH 7.5, 37°C, purified recombinant wild-type enzyme
0.303
N-benzoylmethionine
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pH 7.5, 37°C, purified recombinant wild-type enzyme
0.195
N-benzoylphenylalanine
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pH 7.5, 37°C, purified recombinant wild-type enzyme
0.594
N-benzoylvaline
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pH 7.5, 37°C, purified recombinant wild-type enzyme
0.54
N-[(4-hydroxyphenyl)carbonyl]glycine
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-
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
14.75
4-aminobenzoyl-glutamate
-
pH 7.5, 37°C, purified recombinant wild-type enzyme
1.4 - 98.73
hippurate
2 - 22.12
N-acetyl-L-alanine
0.052 - 10.18
N-acetylglycine
29.38 - 81
N-benzoylalanine
0.17 - 48.72
N-benzoylglutamate
2.6 - 57.13
N-benzoylmethionine
5.95
N-benzoylphenylalanine
-
pH 7.5, 37°C, purified recombinant wild-type enzyme
0.5 - 13.33
N-benzoylvaline
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
120
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purified recombinant wild-type enzyme
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
42000
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4 * 42000, SDS-PAGE
42400
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4 * 42400, recombinant His6-tagged enzyme, SDS-PAGE
170000
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gel filtration
193000
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recombinant His6-tagged enzyme, gel filtration
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SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
tetramer
additional information
-
the enzyme structure shows highly ordered alpha-helix and beta-sheet sequences
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Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
tetrahydral crystals
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.3
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irreversible denaturation
172036
5 - 10
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extremely labile below and above
172035
6 - 8
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stable between, labile in acidic pH
172033
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
-
60% inactivation after 24 h
50
-
stable up to
60
-
rapid inactivation in absence of substrate
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 10-20% loss of activity within few weeks
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant His6-tagged enzyme from Escherichia coli strain M15 by nickel affinity chromatography
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to homogeneity, chromatography techniques
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
gene hipO, expression of His6-tagged enzyme in Escherichia coli strain M15
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D104N
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site-directed mutagenesis, inactive mutant
D76N
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site-directed mutagenesis, the mutant shows over 12fold reduced activity compared to the wild-type enzyme
E134Q
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site-directed mutagenesis, inactive mutant
E135Q
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site-directed mutagenesis, inactive mutant
H161A
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site-directed mutagenesis, inactive mutant
H356A
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site-directed mutagenesis, inactive mutant
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
-
enzymatic determination of Bacillus subtilis siderophore itoic acid and 2,3-dihydroxybenzoic acid
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Show AA Sequence (1145 entries)
Please use the Sequence Search for a specific query.
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LINKS TO OTHER DATABASES (specific for EC-Number 3.5.1.32)
ExplorEnz
ExPASy
KEGG
MetaCyc
SABIO-RK
NCBI: PubMed, Protein, Nucleotide, Structure, Genome, OMIM
IUBMB Enzyme Nomenclature
PROSITE Database of protein families and domains
InterPro (database of protein families, domains and functional sites)