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Information on EC 3.5.1.4 - amidase and Organism(s) Pseudomonas aeruginosa and UniProt Accession P11436
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3.5.1.4 amidaseSpecify your search results
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- 3.5.1.4
-
deamination
- 1-aminocyclopropane-1-carboxylate
- peptidoglycan
- cytidine
- inosine
- esterase
- nitrile
-
siderophore
-
growth-promoting
- rhodococcus
-
hydratase
-
endocannabinoids
- cannabinoids
- iaa
- penicillin
-
hypermutation
- autolysins
-
endophytic
-
indole-3-acetic
-
rhizosphere
- rhizobacteria
- apobec3s
- deoxycytidine
-
editor
-
activation-induced
- porphobilinogen
- palmitoylethanolamide
-
a-to-i
- glucosamine-6-phosphate
- erythropolis
- porphyria
- 2-arachidonoylglycerol
- endolysins
- monoacylglycerol
- ehna
-
rna-specific
-
biofertilizer
-
dihydrolase
- uroporphyrinogens
-
polypeptide-like
- muropeptides
- deoxycytidylate
-
bacteriolytic
- 5-fluorocytosine
- dcmp
-
2\'-deoxycoformycin
-
diphosphohydrolase
- synthesis
- deoxycoformycin
-
pngase
-
ecto-5\'-nucleotidase
- drug development
- biotechnology
- environmental protection
- medicine
- industry
- pharmacology
The taxonomic range for the selected organisms is: Pseudomonas aeruginosa
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Synonyms
amidase, deaminase, acylase, acetamidase, amide hydrolase, acid transferase, n-acylethanolamine acid amidase, n-acylethanolamine-hydrolyzing acid amidase, atfaah, acylamidase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
acid transferase
-
-
-
-
acylamidase
-
-
-
-
acylase
-
-
-
-
amide hydrolase
-
-
-
-
amide transferase
-
-
-
-
amidohydrolase
-
-
-
-
deaminase
-
-
-
-
ester transferase
-
-
-
-
N-acetylaminohydrolase
-
-
-
-
Wide spectrum amidase
-
-
-
-
additional information
-
the enzyme belongs to the thiol nitrilases family
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a monocarboxylic acid amide + H2O = a monocarboxylate + NH3
hydrolysis of aliphatic amide
-
a monocarboxylic acid amide + H2O = a monocarboxylate + NH3
ping-pong mechanism, substitution mechanism
-
a monocarboxylic acid amide + H2O = a monocarboxylate + NH3
the enzyme contains a nitrilase catalytic triad at the bottom of a 13-A deep, funnel-shaped pocket, accessible from the solvent through a narrow neck with 3-A diameter
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of carboxylic acid amide
transfer of amide group
transfer of acyl group
hydrolysis of carboxylic acid amide
-
-
-
-
transfer of amide group
-
-
-
-
transfer of acyl group
-
-
-
-
PATHWAY SOURCE
PATHWAYS
BRENDA
KEGG
-
-, -, -, -, -
SYSTEMATIC NAME
IUBMB Comments
acylamide amidohydrolase
-
CAS REGISTRY NUMBER
COMMENTARY
9012-56-0
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
hydroxylamine + acylamide
acylhydroxamate + NH3
-
acyltransferase activity
-
?
p-nitroacetanilide + H2O
acetic acid + p-nitroaniline
-
mutant T103I
-
-
?
p-nitrophenylacetamide + H2O
p-nitrophenylacetic acid + NH3
-
mutant W138G and T103I-W138G
-
-
?
phenylacetamide + H2O
phenylacetate + NH3
-
mutant W138G and T103I-W138G
-
-
?
additional information
?
-
-
the enzyme catalyses the hydrolysis of small aliphatic amides
-
-
?
additional information
?
-
-
the enzyme catalyses the hydrolysis of small aliphatic amides
-
-
?
additional information
?
-
-
formation of acetohydroxamic acid using acetamide and hydroxylamine as substrates
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
hydroxylamine + acylamide
acylhydroxamate + NH3
-
acyltransferase activity
-
?
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
(2R,3R)-allo-2-methyl-4-oxo-3-oxetanylcarbamic acid tert-butyl ester
-
-
(2S,3R)-2-methyl-4-oxo-3-oxetanylcarbamic acid 5-phenylpentyl ester
-
potent inhibitor, rapidly cleaved in plasma
(2S,3S)-allo-2-methyl-4-oxo-3-oxetanylcarbamic acid tert-butyl ester
-
-
Chloroacetone
-
pH 8.66, 25°C, irreversible strong inhibition at concentrations of 0.1 mM and above, acetamide protects enzyme from inhibition and slowly recovers activity of inactivated enzyme
ethyl 6-[[(3S)-2-oxooxetan-3-yl]carbamoyl]naphthalene-2-carboxylate
-
-
phenyl 6-[[(3S)-2-oxooxetan-3-yl]carbamoyl]naphthalene-2-carboxylate
-
-
Cyanate
-
Ki of strain AI3 is 1.1 mM, Ki of strain AIU1N is 1.8 mM and Ki of strain OUCH4 is 50 mM
Hydroxyurea
-
Ki of strain AI3 is 1.4 mM, Ki of strain AIU1N is 10.8 mM and Ki of strain OUCH4 is 337 mM, regaines full activity within 24 h at 37°C in 0.05 M Tris buffer at pH 7.2
iodoacetamide
-
hydrolase activity kcat: 0.033 per min with 10 mM at pH 7.0 and 25°C, transferase activity 75%, remaining activity less than 2%
iodoacetate
-
kcat: 0.00325 per min with 250 mM at pH 7.0 and 25°C, remaining activity less than 2%
Urea
-
competitive inhibitor at pH 7.2, 90 mM acetamide protectes AI3 amidase against 2.9 mM urea over 2 h, Ki of strain AI3 is 6.6 mM, Ki of strain AIU1N is 41.3 mM and Ki of strain OUCH4 is not known, regaines full activity within 24 h at 37°C in 0.05 M Tris buffer at pH 7.2
additional information
-
no inhibition with 10 mM guanidinium carbonate, 100 mM thiourea, ammonium carbamate at strain AI 3
-
additional information
-
effects of specific monoclonal antibodies on wild-type and mutant enzyme activity
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0049
(2R,3R)-allo-2-methyl-4-oxo-3-oxetanylcarbamic acid tert-butyl ester
Pseudomonas aeruginosa
-
pH 5.0, 37°C, recombinant enzyme
0.1
(2R,3S)-2-methyl-4-oxo-3-oxetanylcarbamic acid benzyl ester
Pseudomonas aeruginosa
-
pH 5.0, 37°C, recombinant enzyme
0.08
(2R,3S)-2-methyl-4-oxo-3-oxetanylcarbamic acid tert-butyl ester
Pseudomonas aeruginosa
-
pH 5.0, 37°C, recombinant enzyme
0.000127
(2S,3R)-2-methyl-4-oxo-3-oxetanylcarbamic acid 5-phenylpentyl ester
Pseudomonas aeruginosa
-
pH 5.0, 37°C, recombinant enzyme
0.001
(2S,3R)-2-methyl-4-oxo-3-oxetanylcarbamic acid benzyl ester
Pseudomonas aeruginosa
-
pH 5.0, 37°C, recombinant enzyme
0.00022
(2S,3R)-2-methyl-4-oxo-3-oxetanylcarbamic acid tert-butyl ester
Pseudomonas aeruginosa
-
pH 5.0, 37°C, recombinant enzyme
0.00019
(2S,3S)-allo-2-methyl-4-oxo-3-oxetanylcarbamic acid tert-butyl ester
Pseudomonas aeruginosa
-
pH 5.0, 37°C, recombinant enzyme
0.0005
(R)-(2-oxo-3-oxetanyl)carbamic acid tert-butyl ester
Pseudomonas aeruginosa
-
pH 5.0, 37°C, recombinant enzyme
0.0492
(R)-1-methyl-3-(2-oxo-3-oxetanyl)urea
Pseudomonas aeruginosa
-
pH 5.0, 37°C, recombinant enzyme
0.0007
(R)-2-oxo-3-oxetanylcarbamic acid benzyl ester
Pseudomonas aeruginosa
-
pH 5.0, 37°C, recombinant enzyme
0.1
(R,S)-2-oxocyclobutylcarbamic acid benzyl ester
Pseudomonas aeruginosa
-
pH 5.0, 37°C, recombinant enzyme
0.00058
(S)-(2-oxo-3-oxetanyl)carbamic acid tert-butyl ester
Pseudomonas aeruginosa
-
pH 5.0, 37°C, recombinant enzyme
0.015
(S)-1-methyl-3-(2-oxo-3-oxetanyl)urea
Pseudomonas aeruginosa
-
pH 5.0, 37°C, recombinant enzyme
0.00296
(S)-2-oxo-3-oxetanylcarbamic acid benzyl ester
Pseudomonas aeruginosa
-
pH 5.0, 37°C, recombinant enzyme
0.1
cyclobutylcarbamic acid benzyl ester
Pseudomonas aeruginosa
-
pH 5.0, 37°C, recombinant enzyme
0.000115
N-[(3S)-2-oxooxetan-3-yl]biphenyl-4-carboxamide
Pseudomonas aeruginosa
-
pH 5.0, 37°C, recombinant enzyme
0.1
N-[(benzyloxy)carbonyl]-D-serine
Pseudomonas aeruginosa
-
pH 5.0, 37°C, recombinant enzyme
0.1
N-[(benzyloxy)carbonyl]-L-serine
Pseudomonas aeruginosa
-
pH 5.0, 37°C, recombinant enzyme
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.0017
-
growth substrate propionamide, 2-chloropropionamide-amidase activity, measured with 80 mM 2-chloropropionamide, strain PAU3
1.5
10.5
7.4
additional information
10.5
-
substrate 4-nitro-acetanilide, strain AI3 25-fold greater activity than wild-type
additional information
-
effects of specific monoclonal antibodies on wild-type and mutant enzyme activity
additional information
-
effects of specific monoclonal antibodies on wild-type and mutant enzyme activity
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
200000
-
gel filtration and sedimentation equilibrium analysis, sedimentation coeffient 10.6 S
221000
224000
240000
35000
36000
38000
38400
38441
78000
240000
-
calculated from measurement of the subunit molecular weight, strain AI3
38441
-
alpha = beta = gamma = 75°, and space group, R3 or R32, quaternary structure determinated by gel filtration and SDS-PAGE, 6 * 38441 deduced from primary structure
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hexamer
additional information
hexamer
-
with most probably 32 symmetry, crystals of both wild-type and C166S amidase grow with identical, rhombohedral morphology. X-ray diffraction analysis established the unit cell dimensions, a = b = c = 84 A
hexamer
-
alpha = beta = gamma = 75°, and space group, R3 or R32, quaternary structure determinated by gel filtration and SDS-PAGE, 6 * 38441 deduced from primary structure
additional information
-
each amidase monomer is formed by a globular four-layer alphabetabetaalpha sandwich domain with an additional 81-residue long C-terminal segment, this wraps arm-in-arm with a homologous C-terminal chain of another monomer, producing a strongly packed dimer, the active homo-hexameric amidase is built grouping three such dimers around a crystallographic 3fold axis, structure analysis and comparisons, overview
additional information
-
epitope binding of specific monoclonal antibodies to the wild-type and T103I mutant enzyme, overview
additional information
-
epitope binding of specific monoclonal antibodies to the wild-type and T103I mutant enzyme, overview
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
high resolution crystallographic structure shows that each amidase monomer is formed by a globular four-layer alphabetbetaalpha sandwich domain with an additional 81-residue long C-terminal segment. This wraps arm-in-arm with a homologous C-terminal chain of another monomer, producing a strongly packed dimer. In the crystal, the biological active homo-hexameric amidase is built grouping three such dimers around a crystallographic 3fold axis
-
purified wild-type enzyme, 25 mg/ml protein in 50 mM Tris-HCl, pH 7.2, containing 5 mM DTT and 1 mM EDTA, hexagonal crystals, X-ray diffraction structure determination and analysis at 1.25-1.4 A resolution
-
purified wild-type enzyme, 25 mg/ml protein in 50 mM Tris-HCl, pH 7.2, containing 5 mM DTT and 1 mM EDTA, hexagonal crystals, X-ray diffraction structure determination and analysis at 1.25-1.4 A resolution, modeling
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
K134R
200fold reduction in activity compared to wild type enzyme
T103I
T103I-W138G
-
mutant with increased instability and less themrmostable
additional information
-
the constitutive mutant strain L10 is isolated by mutagenesis from the wild-type strain 8602
T103I
-
mutant able to use acetanilide as a carbon source, less thermostable
T103I
-
site-directed mutagenesis, binding of specific monoclonal antibodies to the mutant enzyme compared to the wild-type enzyme
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-30°C, 0.05 M Tris buffer, pH 7.2, 50% v/v glycerol, 1 mM 2-mercaptoethanol
-
purified amidase stored in 0.05 M Tris buffer, pH 7.2, containing 1 mM mercaptoethanol and 50%, v/v, glycerol, at -30°C
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
affinity chromatography on an acetamide epoxy-activated Sepharose column
affinity chromatography on epoxy-activated Sepharose 6B-acetamide and gel filtration
-
ammonium sulfate precipitation and dialysis and gel filtration on DEAE-Sephadex A-50
-
recombinant wild-type and mutant enzymes by acetamide affinity chromatography and gel filtration
-
recombinant wild-type and mutant enzymes from Escherichia coli by acetamide affinity chromatography and gel filtration
-
streptomycin sulfate treatment, a heat treatment, ammonium sulfate fractionation and column chromatography on DEAE-Sephadex , yield 30%, purification was 19fold
-
wild-type enzyme by acetamide affinity chromatography and gel filtration
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene amiE, expression in Escherichia coli strain JM109 using plasmid pKK, production method evaluation and optimization, expression of amidase in soluble form and in aggregates in insoluble fractions, overview
-
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
solubilization of amidase aggregates in insoluble fractions with up to 1.5 M, of solubilizing agent such as L-arginine or GdnHCl, followed by washing, centrifugation and dilution
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Nawaz, S.M.; Khan, A.A.; Seng J.E.; Leakey, J.E.; Siitonen, P.H.; Cerniglia, C.E.
Purification and characterization of an amidase from an acrylamide-degrading Rhodococcus sp.
Appl. Environ. Microbiol.
60
3343-3348
1994
Brevibacterium sp., Pseudomonas sp., Pseudomonas aeruginosa, Rhodococcus sp.
Silman, N.J.; Carver, A.M.; Jones, C.W.
Directed evolution of amidase in Methylophilus methylotrophus; purification and properties of amidases from wild-type and mutants strains
J. Gen. Microbiol.
137
169-178
1991
Arthrobacter sp., Methylophilus methylotrophus, Pseudomonas aeruginosa
-
Brown, P.R.; Smyth, M.J.; Clarke, P.H.; Rosemeyer, M.A.
The subunit structure of the aliphatic amidase from Pseudomonas aeruginosa
Eur. J. Biochem.
34
177-187
1973
Pseudomonas aeruginosa, Pseudomonas aeruginosa PAC 142 (L10R), Pseudomonas aeruginosa PAC 111 (C11)
Woods, M.J.; Orsi, B.A.
Selective inhibition and the kinetic mechanism of the aliphatic amidase of Pseudomonas aeruginosa
Biochem. Soc. Trans.
552
1344-1345
1974
Pseudomonas aeruginosa
-
Woods, M.J.; Findlater, J.D.; Orsi, B.A.
Kinetic mechanismen of the aliphatic amidase from Pseudomonas aeruginosa
Biochim. Biophys. Acta
567
225-237
1979
Pseudomonas aeruginosa
Gregoriou, M.; Brown, P.R.
Inhibition of the aliphatic amidase from Pseudomonas aeruginosa by urea and related compounds
Eur. J. Biochem.
96
101-108
1979
Pseudomonas aeruginosa, Pseudomonas aeruginosa AI3
Wyndham, R.C.; Slater, J.H.
A comparative study of acquired amidase activity in Pseudomaonas species
J. Gen. Microbiol.
132
2185-2204
1986
Pseudomonas aeruginosa, Pseudomonas putida, Pseudomonas putida PP3
-
Ambler, R.P.; Auffret, A.D.; Clarke, P.H.
The amino acid sequence of the aliphatic amidase from Pseudomonas aeruginosa
FEBS Lett.
215
285-290
1987
Pseudomonas aeruginosa, Pseudomonas aeruginosa PAC 142 (L10R)
Farnaud, S.; Tata, Renee.; Sohi, M.K.; Wan, P.R.; Sutton, B.J.
Evidence that cysteine-166 is the active-site nucleophile of Pseudomonas aeruginasa amidase: crystallization and preliminary X-ray diffraction analysis of the enzyme
Biochem. J.
340
711-714
1999
Escherichia coli, Pseudomonas aeruginosa, Escherichia coli JM109
Pacheco, R.; Serralheiro, M.L.M.; Karmali, A.; Haris, P.I.
Measuring enzymatic activity of a recombinant amidase using Fourier transform infrared spectroscopy
Anal. Biochem.
322
208-214
2003
Pseudomonas aeruginosa, Pseudomonas aeruginosa 8602
Novo, C.; Farnaud, S.; Tata, R.; Clemente, A.; Brown, P.R.
Support for a three-dimensional structure predicting a Cys-Glu-Lys catalytic triad for Pseudomonas aeruginosa amidase comes from site-directed mutagenesis and mutations altering substrate specificity
Biochem. J.
365
731-738
2002
Pseudomonas aeruginosa (P11436), Pseudomonas aeruginosa
Hollaway, M.R.; Clarke, P.H.; Ticho, T.
Chloroacetone as an active-site-directed inhibitor of the aliphatic amidase from Pseudomonas aeruginosa
Biochem. J.
191
811-826
1980
Pseudomonas aeruginosa, Pseudomonas aeruginosa PAC142
Domingos, A.; Karmali, A.; Brown, P.R.
One-step affinity purification of amidase from mutant strains of Pseudomonas aeruginosa
Biochimie
71
1179-1184
1989
Pseudomonas aeruginosa
Karmali, A.; Pacheco, R.; Tata, R.; Brown, P.
Substitutions of Thr-103-Ile and Trp-138-Gly in amidase from Pseudomonas aeruginosa are responsible for altered kinetic properties and enzyme instability
Mol. Biotechnol.
17
201-212
2001
Pseudomonas aeruginosa
Andrade, J.; Karmali, A.; Carrondo, M.A.; Frazao, C.
Crystallization, diffraction data collection and preliminary crystallographic analysis of hexagonal crystals of Pseudomonas aeruginosa amidase
Acta Crystallogr. Sect. F
63
214-216
2007
Pseudomonas aeruginosa
Andrade, J.; Karmali, A.; Carrondo, M.A.; Frazao, C.
Structure of amidase from Pseudomonas aeruginosa showing a trapped acyl transfer reaction intermediate state
J. Biol. Chem.
282
19598-19605
2007
Pseudomonas aeruginosa
Martins, S.; Karmali, A.; Andrade, J.; Custodio, A.; Serralheiro, M.L.
Characterization of monoclonal antibodies against altered (T103I) amidase from Pseudomonas aeruginosa
Mol. Biotechnol.
30
207-219
2005
Pseudomonas aeruginosa
Martins, S.; Lourenco, S.; Karmali, A.; Serralheiro, M.L.
Monoclonal antibodies recognize conformational epitopes on wild-type and recombinant mutant amidases from Pseudomonas aeruginosa
Mol. Biotechnol.
37
136-145
2007
Pseudomonas aeruginosa, Pseudomonas aeruginosa 8602
Martins, S.; Andrade, J.; Karmali, A.; Serralheiro, M.L.
Screening of suitable immobilized metal chelates for adsorption of monoclonal antibodies against mutant amidase from Pseudomonas aeruginosa
J. Mol. Recognit.
19
340-347
2006
Pseudomonas aeruginosa
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