Information on EC 3.5.1.87 - N-carbamoyl-L-amino-acid hydrolase

for references in articles please use BRENDA:EC3.5.1.87
Word Map on EC 3.5.1.87
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:

The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.5.1.87
-
RECOMMENDED NAME
GeneOntology No.
N-carbamoyl-L-amino-acid hydrolase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
an N-carbamoyl-L-2-amino acid (a 2-ureido carboxylate) + H2O = an L-2-amino acid + NH3 + CO2
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of C-N bond
SYSTEMATIC NAME
IUBMB Comments
N-carbamoyl-L-amino-acid amidohydrolase
This enzyme, along with EC 3.5.1.77 (N-carbamoyl-D-amino-acid hydrolase), EC 5.1.99.5 (hydantoin racemase) and hydantoinase, forms part of the reaction cascade known as the "hydantoinase process", which allows the total conversion of D,L-5-monosubstituted hydantoins into optically pure D- or L-amino acids [3]. The enzyme from Alcaligenes xylosoxidans has broad specificity for carbamoyl-L-amino acids, although it is inactive on the carbamoyl derivatives of glutamate, aspartate, arginine, tyrosine or tryptophan. The enzyme from Sinorhizobium meliloti requires a divalent cation for activity and can hydrolyse N-carbamoyl-L-tryptophan as well as N-carbamoyl L-amino acids with aliphatic substituents [2]. The enzyme is inactive on derivatives of D-amino acids. In addition to N-carbamoyl L-amino acids, the enzyme can also hydrolyse formyl and acetyl derivatives to varying degrees [1,2].
CAS REGISTRY NUMBER
COMMENTARY hide
116412-22-7
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
wild-type and inducer-independent mutant strain RU-ORPN1F
-
-
Manually annotated by BRENDA team
MH602, thermophile bacterium
-
-
Manually annotated by BRENDA team
isolated from industrial waste water, gene atcC
H9B8T5
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
an N-carbamoyl-L-2-amino acid + H2O
an L-2-amino acid + NH3 + CO2
show the reaction diagram
-
-
-
-
?
N-acetyl-L-alanine + H2O
L-alanine + acetate
show the reaction diagram
-
-
-
-
?
N-acetyl-L-methionine + H2O
L-methionine + acetate
show the reaction diagram
-
-
-
-
?
N-acetyl-L-methionine + H2O
L-methionine + CO2
show the reaction diagram
-
-
-
-
?
N-acetyl-L-phenylalanine + H2O
L-phenylalanine + acetate
show the reaction diagram
-
-
-
-
?
N-acetyl-L-tyrosine + H2O
L-tyrosine + acetate
show the reaction diagram
-
-
-
-
?
N-acetyl-L-valine + H2O
L-valine + acetate
show the reaction diagram
-
-
-
-
?
N-carbamoyl-D,L-serine + H2O
L-serine + CO2 + NH3
show the reaction diagram
-
-
-
-
?
N-carbamoyl-DL-2-aminoburyric acid + H2O
L-2-aminobutyric acid + CO2 + NH3
show the reaction diagram
N-carbamoyl-DL-2-aminohexanoic acid + H2O
2-aminohexanoic acid + CO2 + NH3
show the reaction diagram
-
24% of activity with N-carbamoyl-DL-alanine
-
?
N-carbamoyl-DL-alanine + H2O
L-alanine + CO2 + NH3
show the reaction diagram
N-carbamoyl-DL-aminobutyric acid + H2O
L-aminobutyric acid + CO2 + NH3
show the reaction diagram
-
-
-
?
N-carbamoyl-DL-aspartic acid + H2O
DL-aspartic acid + CO2 + NH3
show the reaction diagram
-
relative activity 7.81%
-
-
?
N-carbamoyl-DL-cysteine + H2O
DL-cysteine + CO2 + NH3
show the reaction diagram
-
relative activity 119%
-
-
?
N-carbamoyl-DL-homophenylalanine + H2O
L-homophenylalanine + CO2 + NH3
show the reaction diagram
N-carbamoyl-DL-methionine + H2O
2-aminohexanoic acid + CO2 + NH3
show the reaction diagram
N-carbamoyl-DL-methionine + H2O
L-methionine + CO2 + NH3
show the reaction diagram
N-carbamoyl-DL-norleucine + H2O
L-norleucine + CO2 + NH3
show the reaction diagram
N-carbamoyl-DL-norvaline + H2O
L-norvaline + CO2 + NH3
show the reaction diagram
N-carbamoyl-DL-phenylalanine + H2O
L-phenylalanine + CO2 + NH3
show the reaction diagram
N-carbamoyl-DL-phenylglycine + H2O
L-phenylglycine + CO2 + NH3
show the reaction diagram
-
low activity
-
-
?
N-carbamoyl-DL-serine + H2O
DL-serine + CO2 + NH3
show the reaction diagram
-
relative activity 37.8%
-
-
?
N-carbamoyl-DL-serine + H2O
L-serine + CO2 + NH3
show the reaction diagram
N-carbamoyl-DL-threonine + H2O
L-threonine + CO2 + NH3
show the reaction diagram
N-carbamoyl-DL-valine + H2O
L-valine + CO2 + NH3
show the reaction diagram
N-carbamoyl-glycine + H2O
glycine + CO2 + NH3
show the reaction diagram
N-carbamoyl-L-alanine + H2O
L-alanine + CO2 + NH3
show the reaction diagram
N-carbamoyl-L-asparagine + H2O
L-asparagine + CO2 + NH3
show the reaction diagram
N-carbamoyl-L-cysteine + H2O
L-cysteine + CO2 + NH3
show the reaction diagram
N-carbamoyl-L-glutamic acid + H2O
L-glutamic acid + CO2 + NH3
show the reaction diagram
N-carbamoyl-L-isoleucine + H2O
L-isoleucine + CO2 + NH3
show the reaction diagram
N-carbamoyl-L-leucine + H2O
L-leucine + CO2 + NH3
show the reaction diagram
N-carbamoyl-L-methionine + H2O
L-methionine + CO2 + NH3
show the reaction diagram
N-carbamoyl-L-phenylalanine + H2O
L-phenylalanine + CO2 + NH3
show the reaction diagram
N-carbamoyl-L-phenylglycine + H2O
L-phenylglycine + CO2 + NH3
show the reaction diagram
-
-
-
-
?
N-carbamoyl-L-thienylalanine + H2O
L-thienylalanine + CO2 + NH3
show the reaction diagram
-
-
-
?
N-carbamoyl-L-tryptophan + H2O
L-tryptophan + CO2 + NH3
show the reaction diagram
N-carbamoyl-L-tyrosine + H2O
L-tyrosine + CO2 + NH3
show the reaction diagram
N-carbamoyl-L-valine + H2O
L-valine + CO2 + NH3
show the reaction diagram
N-formyl-DL-2-aminobutyric acid + H2O
L-2-aminobutyric acid + CO2
show the reaction diagram
-
-
-
-
?
N-formyl-DL-alanine + H2O
L-alanine + CO2
show the reaction diagram
N-formyl-DL-aminobutyric acid + H2O
L-aminobutyric acid + CO2
show the reaction diagram
-
-
-
?
N-formyl-DL-ethionine + H2O
L-ethionine + CO2
show the reaction diagram
N-formyl-DL-homophenylalanine + H2O
L-homophenylalanine + CO2
show the reaction diagram
N-formyl-DL-leucine + H2O
L-leucine + CO2
show the reaction diagram
-
5.2% of activity with N-carbamoyl-DL-alanine
-
?
N-formyl-DL-methionine + H2O
L-methionine + CO2
show the reaction diagram
N-formyl-DL-norleucine + H2O
L-norleucine + CO2
show the reaction diagram
N-formyl-DL-norvaline + H2O
L-norvaline + CO2
show the reaction diagram
N-formyl-DL-phenylalanine + H2O
L-phenylalanine + CO2
show the reaction diagram
-
-
-
?
N-formyl-DL-phenylglycine + H2O
L-phenylglycine + CO2
show the reaction diagram
N-formyl-DL-tryptophan + H2O
L-tryptophan + CO2
show the reaction diagram
N-formyl-L-alanine + H2O
L-alanine + CO2
show the reaction diagram
-
-
-
-
?
N-formyl-L-methionine + H2O
L-methionine + CO2
show the reaction diagram
-
-
-
-
?
N-formyl-L-methionine + H2O
L-methionine + HCOOH
show the reaction diagram
-
-
-
-
?
N-formyl-L-phenylalanine + H2O
L-phenylalanine + CO2
show the reaction diagram
-
-
-
-
?
N-formyl-L-tyrosine + H2O
L-tyrosine + CO2
show the reaction diagram
-
-
-
-
?
N-formyl-L-valine + H2O
L-valine + CO2
show the reaction diagram
-
-
-
-
?
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
an N-carbamoyl-L-2-amino acid + H2O
an L-2-amino acid + NH3 + CO2
show the reaction diagram
-
-
-
-
?
N-carbamoyl-DL-methionine + H2O
L-methionine + CO2 + NH3
show the reaction diagram
N-carbamoyl-glycine + H2O
glycine + CO2 + NH3
show the reaction diagram
-
-
-
-
?
N-carbamoyl-L-alanine + H2O
L-alanine + CO2 + NH3
show the reaction diagram
-
-
-
-
?
N-carbamoyl-L-cysteine + H2O
L-cysteine + CO2 + NH3
show the reaction diagram
N-carbamoyl-L-methionine + H2O
L-methionine + CO2 + NH3
show the reaction diagram
N-formyl-DL-methionine + H2O
L-methionine + CO2
show the reaction diagram
additional information
?
-
-
strictly specific for the L-form of N-carbamoyl-amino acids as substrates and exhibits high activity in the hydrolysis of N-carbamoyl-L-cysteine as substrate
-
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
similar enzyme activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme and HQSA pretreated enzyme compared to activity without addition of any effectors
Cu2+
-
slightly activated activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme and HQSA pretreated enzyme compared to activity without addition of any effectors
Mg2+
-
activates slightly
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
-
54% inhibition at 2 mM
2-mercaptoethanol
-
10 mM causes 29% inhibition, suggesting that no cysteine residue is crucial for enzyme activity, although they might have a structural role
4-chloromercuribenzoic acid
-
-
4-hydroxymercuribenzoate
-
complete inhibition at 1 mM
5,5'-dithiobis(2-nitrobenzoic acid)
5,5'-dithiobis-(2-nitrobenzoic acid)
-
specific activity 0.39, relative activity 58%
8-hydroxyquinoline
-
2 mM, 65% inhibition
8-Hydroxyquinoline-5-sulfonic acid
-
10 mM, decreases the activity to 13%
alpha,alpha'-dipyridyl
-
2.5 mM, 26% inhibition
ATP
-
0.5 mM, approximately 50% inhibition, 1.5 mM, approximately 85% inhibition
beta-mercaptoethanol
-
specific activity 0.44, relative activity 66%
Cd2+
-
2 mM, 86% inhibition
Cs+
-
2 mM, slight inhibition
diisopropyl fluorophosphate
-
5 mM, 18% inhibition
dithiothreitol
-
10 mM causes 41% inhibition, suggesting that no cysteine residue is crucial for enzyme activity, although they might have a structural role
Fe2+
-
2 mM, strong inhibition
iodoacetate
-
2 mM, 43% inhibitiion
iodoacetic acid
K+
-
2 mM, specific activity 0.56, relative activity 84%
Mercuric chloride
-
complete inhibition at 1 mM
Mg2+
-
slightly inhibited activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme and HQSA pretreated enzyme
N-bromosuccinimide
-
complete inhibition at 1 mM
N-ethyl-maleimide
N-ethylmaleimide
-
2 mM, 100% inhibitiion
NaAsO2
-
5 mM, 20% inhibition
o-phenanthroline
-
10 mM, 47% inhibition
p-chloromercuribenzoic acid
phenylhydrazine
-
5 mM, 48% inhibition
phenylmethylsulfonyl fluoride
-
5 mM, 31% inhibition
sodium propionate
-
10 mM, 20% inhibition, competitive inhibition of N-carbamoyl-L-alanine hydrolysis
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Fe2+
-
2 mM, specific activity 2.03, relative activity 303%
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
36.12
N-acetyl-L-alanine
-
pH 7.5, 65°C, preincubation with CoCl2
5.5 - 12.66
N-acetyl-L-methionine
12.9
N-acetyl-L-phenylalanine
-
pH 7.5, 65°C, preincubation with CoCl2
116
N-acetyl-L-tyrosine
-
pH 7.5, 65°C, preincubation with CoCl2
13.92
N-acetyl-L-valine
-
pH 7.5, 65°C, preincubation with CoCl2
0.85
N-carbamoyl-DL-2-aminohexanoic acid
-
-
3.2
N-carbamoyl-DL-methionine
-
-
10
N-carbamoyl-DL-serine
-
-
0.94 - 4.19
N-carbamoyl-L-alanine
1.5
N-carbamoyl-L-asparagine
-
-
4.91
N-carbamoyl-L-cysteine
-
determined at 40°C for 20 min at pH 8.0 after preincubation with 2 mM Ni2+
51.23
N-carbamoyl-L-glutamic acid
-
determined at 40°C for 20 min at pH 8.0 after preincubation with 2 mM Ni2+
0.31
N-carbamoyl-L-isoleucine
-
-
0.86
N-carbamoyl-L-leucine
-
-
0.2 - 17.8
N-carbamoyl-L-methionine
0.92 - 5.82
N-carbamoyl-L-phenylalanine
0.65
N-carbamoyl-L-tryptophan
-
determined at 40°C for 20 min at pH 8.0 after preincubation with 2 mM Ni2+
4.8 - 12.99
N-carbamoyl-L-tyrosine
0.4 - 34.47
N-carbamoyl-L-valine
6.6
N-Carbamoylglycine
-
-
16
N-formyl-DL-alanine
-
-
5
N-formyl-L-alanine
-
pH 7.5, 65°C, preincubation with CoCl2
9.77 - 12.9
N-formyl-L-methionine
13.82
N-formyl-L-phenylalanine
-
pH 7.5, 65°C, preincubation with CoCl2
23.78
N-Formyl-L-tyrosine
-
pH 7.5, 65°C, preincubation with CoCl2
6.75
N-formyl-L-valine
-
pH 7.5, 65°C, preincubation with CoCl2
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
27.86
N-acetyl-L-alanine
-
pH 7.5, 65°C, preincubation with CoCl2
2.26
N-acetyl-L-methionine
-
pH 7.5, 65°C, preincubation with CoCl2
0.01
N-acetyl-L-phenylalanine
-
pH 7.5, 65°C, preincubation with CoCl2
0.08
N-acetyl-L-tyrosine
-
pH 7.5, 65°C, preincubation with CoCl2
0.08
N-acetyl-L-valine
-
pH 7.5, 65°C, preincubation with CoCl2
7.56
N-Carbamoyl-glycine
-
pH 9.0, 50°C
6.11 - 48.96
N-carbamoyl-L-alanine
16.5
N-carbamoyl-L-cysteine
-
pH 9.0, 50°C
0.9 - 5.32
N-carbamoyl-L-methionine
0.16
N-carbamoyl-L-phenylalanine
-
pH 7.5, 65°C, preincubation with CoCl2
16.1
N-carbamoyl-L-thienylalanine
-
-
5.1
N-carbamoyl-L-tryptophan
-
-
0.18 - 6.5
N-carbamoyl-L-tyrosine
0.84
N-carbamoyl-L-valine
-
pH 7.5, 65°C, preincubation with CoCl2
5
N-formyl-DL-tryptophan
-
-
293.5
N-formyl-L-alanine
-
pH 7.5, 65°C, preincubation with CoCl2
839.1
N-formyl-L-methionine
-
pH 7.5, 65°C, preincubation with CoCl2
12.12
N-formyl-L-phenylalanine
-
pH 7.5, 65°C, preincubation with CoCl2
1.46
N-Formyl-L-tyrosine
-
pH 7.5, 65°C, preincubation with CoCl2
27.26
N-formyl-L-valine
-
pH 7.5, 65°C, preincubation with CoCl2
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
771.3
N-acetyl-L-alanine
-
pH 7.5, 65°C, preincubation with CoCl2
178.5
N-acetyl-L-methionine
-
pH 7.5, 65°C, preincubation with CoCl2
0.39
N-acetyl-L-phenylalanine
-
pH 7.5, 65°C, preincubation with CoCl2
0.69
N-acetyl-L-tyrosine
-
pH 7.5, 65°C, preincubation with CoCl2
5.96
N-acetyl-L-valine
-
pH 7.5, 65°C, preincubation with CoCl2
1.89
N-Carbamoyl-glycine
-
pH 9.0, 50°C
2.57 - 11680
N-carbamoyl-L-alanine
40.2
N-carbamoyl-L-cysteine
-
pH 9.0, 50°C
5.9 - 1834
N-carbamoyl-L-methionine
27.49
N-carbamoyl-L-phenylalanine
-
pH 7.5, 65°C, preincubation with CoCl2
13.86
N-carbamoyl-L-tyrosine
-
pH 7.5, 65°C, preincubation with CoCl2
255.3
N-carbamoyl-L-valine
-
pH 7.5, 65°C, preincubation with CoCl2
58700
N-formyl-L-alanine
-
pH 7.5, 65°C, preincubation with CoCl2
85880
N-formyl-L-methionine
-
pH 7.5, 65°C, preincubation with CoCl2
877
N-formyl-L-phenylalanine
-
pH 7.5, 65°C, preincubation with CoCl2
61.4
N-Formyl-L-tyrosine
-
pH 7.5, 65°C, preincubation with CoCl2
4039
N-formyl-L-valine
-
pH 7.5, 65°C, preincubation with CoCl2
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.66
-
N-carbamoyl-L-methionine
2.91
-
pH 8.0, 50°C, immobilized enzyme
10
-
pH 8.0, 50°C, free enzyme
15.9
-
protein recovered from inclusion bodies
21.5
-
at pH 7.5 and 42°C
33.34
-
activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme, pH 7.5, 65°C
40.2
-
120% relative activity of 8-hydroxyquinoline-5-sulfonic acid (HQSA) pretreated enzyme compared to activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme, pH 7.5, 65°C
65.69
-
Mn2+, 197% relative activity of 8-hydroxyquinoline-5-sulfonic acid (HQSA) pretreated enzyme compared to activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme without addition of any effectors, pH 7.5, 65°C
95.67
-
Mn2+, 286.93% relative activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme compared to activity without addition of any effectors, pH 7.5, 65°C
111.3
-
Ni2+, 333.96% relative activity of 8-hydroxyquinoline-5-sulfonic acid (HQSA) pretreated enzyme compared to activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme without addition of any effectors, pH 7.5, 65°C
114
-
Ni2+, 341.94% relative activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme compared to activity without addition of any effectors, pH 7.5, 65°C
145
-
purified enzyme, pH 7.5, 40°C
179
-
Co2+, 536.91% relative activity of 8-hydroxyquinoline-5-sulfonic acid (HQSA) pretreated enzyme pretreated enzyme compared to activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme without addition of any effectors, pH 7.5, 65°C
221.4
-
Co2+, 663.89% relative activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme compared to activity without addition of any effectors, pH 7.5, 65°C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8 - 8.3
-
N-carbamoyl-L-valine hydrolysis
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 11
-
upon immobilization the operational pH range and temperature range are markedly broadened, ca. 35% relative activity at pH 5 for immoblized enzyme, 85% for free enzyme. At pH 11 ca. 60% relative activity for free enzyme and 50% for the immobilized enzyme is observed
5.5 - 9
7 - 11
-
enzyme activity significantly decreases below, enzyme is highly active up to pH 11.0
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40 - 50
-
immobilized enzyme
45 - 70
-
biocatalyst system, optimum temperature ranges are 45-55°C for N-formyl-amino acids and 55-70°C for N-carbamoyl-derivatives
60
-
activity assay
65
-
maximal activity for hydrolysis of N-acetyl and N-formyl-amino acids and N-carbamoyl-amino acids
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 75
-
activity range, profile overview
25 - 50
-
-
40 - 80
-
upon immobilization the operational pH range and temperature range are markedly broadened, ca. 85% relative activity at 40°C for immoblized enzyme, 100% for free enzyme. At 80°C ca. 35% relative activity for free enzyme and 40% for the immobilized enzyme is observed
40 - 70
-
activity range
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.45
-
isoelectric focusing
PDB
SCOP
CATH
ORGANISM
UNIPROT
Burkholderia ambifaria (strain MC40-6)
Burkholderia cenocepacia (strain ATCC BAA-245 / DSM 16553 / LMG 16656 / NCTC 13227 / J2315 / CF5610)
Burkholderia multivorans (strain ATCC 17616 / 249)
Burkholderia vietnamiensis (strain G4 / LMG 22486)
Paraburkholderia phymatum (strain DSM 17167 / CIP 108236 / LMG 21445 / STM815)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
42000
-
SDS-PAGE
44300
-
3 * 44300, SDS-PAGE
44860
-
molecular mass deduced from the amino acid sequence
47350
-
calculated from sequence
48000
H9B8T5
x * 48000, recombinant enzyme, SDS-PAGE
65000
-
2 * 65000, SDS-PAGE
93000
-
gel filtration
109000
-
recombinant enzyme expressed in E. coli, gel filtration
132000
-
gel filtration
134000
-
gel filtration
170000
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homotrimer
tetramer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging-drop vapour diffusion. X-ray data are collected to a resolution of 2.75 A. The crystals belong to space group P21212, with unit-cell parameters a = 103.2, b = 211.7, c = 43.1 A and two subunits in the asymmetric unit
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 9.5
-
80% of initial activity after 30 min and 30°C
288945
8
-
45°C, the bienzymatic system retains almost total operability after 24 h of incubation at pH 8.0, overview
735201
additional information
-
the immobilized enzyme exhibits higher pH stability than the free enyzme
713443
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 55
-
no loss of activity after 6 d at 37°C, 50% loss of activity after 2 h at 50°C
25
-
activity is lost after 60 min at 25°C
30 - 35
-
no loss of activity after 30 min at pH 7.4, 30% activity lost after 30 min at 35°C
30 - 70
-
immobilized enzyme, in 100 mM borate-HCl, pH 8.0, 18 h, conversion rates decrease at temperatures over 55°C and are lost completely at over 70°C, for the immobilized enzyme system in production of optically pure L-amino acids, overview
45
-
pH 8.0, the bienzymatic system retains almost total operability after 24 h of incubation at 45°C, overview
50
-
purified enzyme, 60 min, about 86% activity remaining, half-life is 180 min
65 - 70
-
thermal stability studies performed on BsLcar show that maximum activity gradually decreases at temperatures over 65°C for 60 min, with 60% activity remaining after incubation at 70°C for 60 min
80
-
incubation at 80°C results in a complete loss of both, the free and immobilized enzyme
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
the hydrolysis acitvity of the immobilized L-N-carbamoylase drops by 40% after 14 cycles
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, the immobilized enzyme retaines approximately 50% of its activity after 5 months, while the free enzyme loses the complete enzyme activity with a half-life of 12 days
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; recombinant His6-tagged enzyme, metal affinity chromatography
-
a one-step purification procedure of the recombinant L-N-carbamoylase fused to the HIS6-tag is employed using immobilized cobalt affinity chromatography
-
ammonium sulfate, phenyl-Sepharose CL-4B, partially purified
-
ammonium sulfate, phenyl-Sepharose, DEAE-cellulose, phenyl-Sepharose, Sephadex G-100
-
anion-exchange chromatography, hydrophobic interaction chromatography hydroxyapatite chromatography used for purification of native expressed protein
-
DEAE-Sephacel, ammonium sulfate, phenyl-Sepharose CL-4B, Mono Q, Superose 12 HR
-
native enzyme 60fold to homogeneity by heat treatment at 70°C for 2 min, anion exchange chromatography of the supernatant, hydrophobic interaction chromatography, ultrafiltration, and gel filtration
-
one-step purification procedure of SmLcar mutants fused to the His6 tag employing cobalt affinity chromatography
-
purified 58fold to homogeneity with a yield of 16.1% by three steps of column chromatography
-
purified to over 95% purity
-
purity of 95%, purified enzyme is used for analytical analysis and enzyme immobilization
-
recombinant enzyme by cobalt affinity chromatography, ultrafiltration and dialysis
-
recombinant enzyme from Escherichia coli
H9B8T5
streamline DEAE, Mono Q
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Asp6 and His6-tagged enzyme, expression in Escherichia coli
-
expressed in Escherichia coli
expressed in Escherichia coli BL21
expression in Escherichia coli
gene atcC, organized in the L-cysteine synthesis gene cluster, DNA and amino acid sequence determination and analysis, recombinant expression in Escherichia coli
H9B8T5
gene Bslcar, recombinant expression
-
gene Bslcar, recombinant overexpression in Escherichia coli strain BL21 using the pJAVI80Rha plasmid
-
L-N-carbamoylase gene of Sinorhizobium meliloti was cloned into the pBluescipt II SK (+) vector for expression in Escherichia coli
-
NCC amidohydrolase gene is cloned into DH5alpha and pet15b vectors for protein expression in Escherichia coli
-
the L-N-carbamoylase gene is amplified by PCR and cloned into the pBSK vector for expression of the protein in Escherichia coli BL21(DE3) cells
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E132D
-
SmLcar mutant
E132H
-
SmLcar mutant
E132Q
-
SmLcar mutant
H230A
-
SmLcar mutant
N279A
-
SmLcar mutant
R292I
-
SmLcar mutant
R292M
-
SmLcar mutant
additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
protein from inclusion bodies is solubilized, recovered and purified by metal affinity chromatography
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
-
production of optically pure L-amino acids by an enzymatic method named hydantoinase process
synthesis
additional information
Show AA Sequence (1338 entries)
Please use the Sequence Search for a specific query.