Information on EC 3.5.2.5 - allantoinase

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

EC NUMBER
COMMENTARY
3.5.2.5
-
RECOMMENDED NAME
GeneOntology No.
allantoinase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
(S)-allantoin + H2O = allantoate
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis
-
-
-
-
hydrolysis
B5L363, -
-
hydrolysis
P77671
-
PATHWAY
KEGG Link
MetaCyc Link
allantoin degradation to ureidoglycolate I (urea producing)
-
allantoin degradation to ureidoglycolate II (ammonia producing)
-
Metabolic pathways
-
Microbial metabolism in diverse environments
-
Purine metabolism
-
SYSTEMATIC NAME
IUBMB Comments
(S)-allantoin amidohydrolase
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
AaALN
Q171F8
-
AaALN
Aedes aegypti NH-Rockefeller
Q171F8
-
-
allantoin amidohydrolase
-
-
allantoinase
Q171F8
-
allantoinase
Aedes aegypti NH-Rockefeller
Q171F8
-
-
allantoinase
P77671
-
allantoinase
B5L363
-
allantoinase
-
-
ALLase
C6EKV4
-
ALN
I1M259
-
Fe-allantoinase
P77671
-
IsoI
-
45000 Da enzyme
IsoII
-
42000 Da enzyme
metal-dependent allantoinase
P77671
-
metal-independent allantoinase
B5L363
-
zinc-amended allantoinase
P77671
-
CAS REGISTRY NUMBER
COMMENTARY
9025-20-1
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
NH-Rockefeller strain
UniProt
Manually annotated by BRENDA team
Aedes aegypti NH-Rockefeller
NH-Rockefeller strain
UniProt
Manually annotated by BRENDA team
; gene aln
-
-
Manually annotated by BRENDA team
one copy gene AtALN
-
-
Manually annotated by BRENDA team
pigeonpea
-
-
Manually annotated by BRENDA team
isoform ALN1
UniProt
Manually annotated by BRENDA team
isoform ALN4
UniProt
Manually annotated by BRENDA team
sunflower
-
-
Manually annotated by BRENDA team
; cv. Great Northern, two isozymes IsoI and IsoII
-
-
Manually annotated by BRENDA team
bullfrog; bull frog
-
-
Manually annotated by BRENDA team
castor bean
-
-
Manually annotated by BRENDA team
a nonureide-type legume black locust, gene RpALN
Uniprot
Manually annotated by BRENDA team
male and female brown trout
-
-
Manually annotated by BRENDA team
corn
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(S)-allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
(S)-allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
(S)-allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
(S)-allantoin + H2O
allantoate
show the reaction diagram
-
-
-
?
(S)-allantoin + H2O
allantoate
show the reaction diagram
P77671
-
-
-
?
(S)-allantoin + H2O
allantoate
show the reaction diagram
C6EKV4
-
-
-
?
(S)-allantoin + H2O
allantoate
show the reaction diagram
-
zinc enzyme uses only the S-isomer, cobalt-enzyme prefers S-isomer but also hydrolyses R-isomer
-
?
(S)-allantoin + H2O
allantoate
show the reaction diagram
P77671
final step of ureide pathway
-
-
?
(S)-allantoin + H2O
allantoate
show the reaction diagram
B5L363, -
0.2 mM, assay at pH 7.6, 25C, 10 min
-
-
r
allantoic acid
(S)-allantoin + H2O
show the reaction diagram
B5L363, -
-
-
-
r
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
ir
allantoin + H2O
allantoate
show the reaction diagram
Q6S4R9, -
-
-
-
ir
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
key enzyme in the biogenesis and catabolism of ureide compounds in plants, pathway overview
-
-
ir
allantoin + H2O
allantoate
show the reaction diagram
Q6S4R9, -
key enzyme in the biogenesis and catabolism of ureide compounds in plants, pathway overview
-
-
ir
allantoin + H2O
allantoate
show the reaction diagram
-
step in the purine degradation pathway producing nitrogen waste for excretion
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
step in the purine degradation pathway, changes during the annual reproductive cycle of the fish
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
a key reaction step in the biosynthesis and degradation of ureides, overview
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
development of a enzyme cycling method for measuring allantoin concentrations in human serum involving allantoinase, glutamine synthetase II, EC 6.3.1.2, allantoate amidohydrolase, EC 3.5.3.9, and NAD synthetase, EC 6.3.1.5, followed by action of glucose dehydrogenase, EC 1.1.1.47, and diaphorase, EC 1.6.99.2, in the presence of glucose and 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, overview, optimal reaction for allantoinase at 0.07 mM allantoin concentration
-
-
?
additional information
?
-
Q6S4R9, -
the enzyme is inducible by allantoin and is upregulated in spring in trunk bark, the enzyme might be regulated by exogenous nitrogen conditions
-
-
-
additional information
?
-
-
the enzyme might be regulated by exogenous nitrogen conditions
-
-
-
additional information
?
-
-
no activity with hydantoin and 5-bromouracil
-
-
-
additional information
?
-
C6EKV4
no substrate: hydantoin, dihydrouracil, phthalimide, dihydroorotate, and 3-imonoisoindolinone
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(S)-allantoin + H2O
allantoate
show the reaction diagram
P77671
final step of ureide pathway
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
-
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
key enzyme in the biogenesis and catabolism of ureide compounds in plants, pathway overview
-
-
ir
allantoin + H2O
allantoate
show the reaction diagram
Q6S4R9, -
key enzyme in the biogenesis and catabolism of ureide compounds in plants, pathway overview
-
-
ir
allantoin + H2O
allantoate
show the reaction diagram
-
step in the purine degradation pathway producing nitrogen waste for excretion
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
step in the purine degradation pathway, changes during the annual reproductive cycle of the fish
-
-
?
allantoin + H2O
allantoate
show the reaction diagram
-
a key reaction step in the biosynthesis and degradation of ureides, overview
-
-
?
additional information
?
-
Q6S4R9, -
the enzyme is inducible by allantoin and is upregulated in spring in trunk bark, the enzyme might be regulated by exogenous nitrogen conditions
-
-
-
additional information
?
-
-
the enzyme might be regulated by exogenous nitrogen conditions
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Mn2+
-
Mn2+ activates the enzyme by 25%
Mn2+
-
at 0.5-0.8 mM MnSO4 the activity incrases by about 70%
Zn2+
P77671
required
additional information
-
metalloenzyme
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-mercaptoethanol
-
5 mM
4-hydroxymercuribenzoate
-
inactivation, prevented by DTT
5-amino-4-imidazole-carboxamide hydrochloride
B5L363, -
-
Ca2+
-
62% residual activity at 1 mM for IsoI and 34% residual activity at 1 mM for IsoII; inhibition of isozymes IsoI and IsoII
Co2+
-
40% residual activity at 1 mM for IsoI and 30% residual activity at 1 mM for IsoII; inhibition of isozymes IsoI and IsoII
Cu2+
-
14% residual activity at 1 mM for IsoI and 8% residual activity at 1 mM for IsoII; high inhibition of isozymes IsoI and IsoII
diethyl dicarbonate
-
89% residual activity at 10 mM for IsoI and 90% residual activity at 10 mM for IsoII
dithiothreitol
-
1 mM
dithiothreitol
-
-
dithiothreitol
-
competitive
EDTA
-
49% residual activity at 1 mM for IsoI and 40% residual activity at 1 mM for IsoII; inhibition of isozymes IsoI and IsoII, Fe2+ and Mn2+ restore the enzyme activity partially, while Ca2+ restores it completely
Fe2+
-
75% residual activity at 1 mM for IsoI and 58% residual activity at 1 mM for IsoII; inhibition of isozymes IsoI and IsoII
-
hydantoin
B5L363, -
-
iodoacetic acid
-
16% residual activity at 5 mM for IsoI and 13% residual activity at 5 mM for IsoII
Mg2+
-
44% residual activity at 1 mM for IsoI and 49% residual activity at 1 mM for IsoII; inhibition of isozymes IsoI and IsoII
N-ethylmaleimide
-
-
N-ethylmaleimide
-
54% residual activity at 5 mM for IsoI and 67% residual activity at 5 mM for IsoII
Na+
-
51% residual activity at 1 mM for IsoI and 43% residual activity at 1 mM for IsoII; inhibition of isozymes IsoI and IsoII
p-chloromercuribenzene sulfonic acid
-
-
p-chloromercuribenzoate
-
-
p-hydroxymercuribenzoate
-
10% residual activity at 0.5 mM for IsoI and 15% residual activity at 0.5 mM for IsoII
Zn2+
-
29% residual activity at 1 mM for IsoI and 38% residual activity at 1 mM for IsoII; inhibition of isozymes IsoI and IsoII
Mn2+
-
81% residual activity at 1 mM for IsoI and 51% residual activity at 1 mM for IsoII; inhibition of isozymes IsoI and IsoII
additional information
-
the enzyme is inhibited by sulfhydryl-reactive agents
-
additional information
-, Q171F8
in fat body decreased expression after blood meal
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
beta-mercaptoethanol
-
14% increase of activity at 0.1 mM for IsoI and 11% increase of activity at 1 mM for IsoII
diethyl dicarbonate
-
63% increase of activity at 0.1 mM for IsoI and 38% increase of activity at 1 mM for IsoII
phenyl phosphoramidate
-
4% increase of activity at 1 mM for IsoI and 10% increase of activity at 1 mM for IsoII
dithiothreitol
-
20% increase of activity at 1 mM for IsoI and% increase of activity at 0.1 mM for IsoII
additional information
Q6S4R9, -
the enzyme is inducible by allantoin and is upregulated in spring in trunk bark
-
additional information
P77671
inactive in absence of zinc
-
additional information
-, Q171F8
in malpighian tubule increased expression after blood meal
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.26
-
(S)-allantoin
B5L363, -
-
5.5
-
(S)-allantoin
C6EKV4
Co2+-activated enzyme, pH 8.0, 25C
6.2
-
(S)-allantoin
C6EKV4
Mn2+-activated enzyme, pH 8.0, 25C
7.3
-
(S)-allantoin
C6EKV4
Ni2+-activated enzyme, pH 8.0, 25C
9.6
-
(S)-allantoin
C6EKV4
Zn2+-activated enzyme, pH 8.0, 25C
17
-
(S)-allantoin
-
pH 7.4, 37C, Zn2+-amended culture
19.5
-
(S)-allantoin
-
pH 7.4, 37C, Co2+-amended culture
28.6
-
(S)-allantoin
P77671
wild-type allantoinase, supplementation with zinc acetate
37.5
-
(S)-allantoin
P77671
selenomethionine-substituted wild-type allantoinase, no supplementation with zinc acetate
53.8
-
(S)-allantoin
P77671
mutant S288D, supplementation with zinc acetate
62.8
-
(S)-allantoin
P77671
mutant R215K, supplementation with zinc acetate
79.9
-
(S)-allantoin
-
pH 7.4, 37C, Ni2+-amended culture
91.3
-
(S)-allantoin
P77671
mutant S288V, supplementation with zinc acetate
2.4
-
Allantoic acid
B5L363, -
-
0.059
-
allantoin
-
pH 7.8, 40C, isozyme IsoI, different methods
0.066
-
allantoin
-
pH 7.8, 40C, isozyme IsoII, different methods
2.56
-
allantoin
-
-
6.6
-
allantoin
-
-
10
-
allantoin
-
-
13.3
-
allantoin
-
-
13.8
-
allantoin
-
-
additional information
-
additional information
-
Michaelis-Menten kinetics
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
201.7
-
allantoin
-
pH 7.8, 40C, isozyme IsoII, different methods
420
-
allantoin
-
pH 7.8, 40C, isozyme IsoI, different methods
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.237
-
dithiothreitol
-
pH 7.4, 37C, Co2+-amended culture
0.795
-
dithiothreitol
-
pH 7.4, 37C, Zn2+-amended culture
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.002
-
-
-
0.006
-
-
-
0.03
-
-
-
0.037
-
-
-
0.08
-
-
pH 7.5
0.5
-
-
pH 7.5
1
-
-
wild-type seedlings grown on Murashige Skoog medium
1.3
-
C6EKV4
Ni2+-activated enzyme, pH 8.0, 25C
2
-
-
crude extract
2.6
-
C6EKV4
Zn2+-activated enzyme, pH 8.0, 25C
5
-
Q6S4R9, -
seedlings grown on Murashige Skoog medium
8
-
C6EKV4
Co2+-activated enzyme, pH 8.0, 25C
10
-
-
wild-type seedlings grown on ALN medium
76.1
-
-
pH 7.4, 37C, Zn2+-amended culture
88.2
-
C6EKV4
Mn2+-activated enzyme, pH 8.0, 25C
295
-
-
purified isozyme IsoII
400
-
-
after 200fold purification
410
-
-
pH 7.4, 37C, Co2+-amended culture
563
-
-
purified isozyme IsoI
additional information
-
-
activity of isozymes in different bean tissues and during development, overview
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.5
7.6
-
-
7.5
7.7
-
-
7.5
-
-
-
7.5
-
-
assay at
7.6
-
B5L363, -
assay at
7.7
-
-
different methods
7.8
-
-
different methods; IsoI
8.1
-
-
IsoII
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
-
assay at
25
-
B5L363, -
assay at
55
-
C6EKV4
Mn2+-activited enzyme
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
Aedes aegypti NH-Rockefeller
-
-
-
Manually annotated by BRENDA team
-
developing, high enzyme expression level; highest activity
Manually annotated by BRENDA team
-
high activity independent of the time in the year
Manually annotated by BRENDA team
-
; IsoI is the predominant form in all the tissues analysed except leaves, where most of the allantoinase activity corresponds to IsoII enzyme
Manually annotated by BRENDA team
-
in shoots and leaves from plants using symbiotically fixed nitrogen as the sole nitrogen source, ureide levels are roughly equivalent to those of nitrate-supported plants during the whole vegetative stage, but exhibit a sudden increase at the onset of flowering. This increase is accompanied by increases in allantoinase gene expression and enzyme activity
Manually annotated by BRENDA team
-
the enzyme acivity is higher in female liver in May and decreases during vitellogenesis, similar in male livers
Manually annotated by BRENDA team
Aedes aegypti NH-Rockefeller
-
-
-
Manually annotated by BRENDA team
-
; the amount of IsoII is very low in roots
Manually annotated by BRENDA team
Q6S4R9, -
axis, cotyledon, and hypocotyl, the enzyme expression is differentially regulated in tissues during seed germination and seedling development, no activity in seedling roots
Manually annotated by BRENDA team
-
in shoots and leaves from plants using symbiotically fixed nitrogen as the sole nitrogen source, ureide levels are roughly equivalent to those of nitrate-supported plants during the whole vegetative stage, but exhibit a sudden increase at the onset of flowering. This increase is accompanied by increases in allantoinase gene expression and enzyme activity
Manually annotated by BRENDA team
Q6S4R9, -
wood, high expression level in bark/cambial region, which is upregulated n spring, but no expression in the sapwood-heartwood transition zone
Manually annotated by BRENDA team
additional information
-
activity of purine catabolism enzymes during the reproductive cycle of male and female brown trout, overview
Manually annotated by BRENDA team
additional information
-
developmental expression analysis, no activity in pupae and eggs
Manually annotated by BRENDA team
additional information
Q6S4R9, -
expression pattern analysis, no activity in roots
Manually annotated by BRENDA team
additional information
-
tissue distribution, expression especially during fertilization, ubiquitous expression of isozymes in bean tissues, one isozyme is the main form in leaves and the other isozyme is the main form in roots, overview
Manually annotated by BRENDA team
additional information
I1M259, I1MEH3
present in all tissues of germinating seedling and in vegetative tissues from 45 day old plants. Isoforms ALN1 and ALN2 are consistently expressed at higher levels than ALN3 and ALN4 in all samples; present in all tissues of germinating seedling and in vegetative tissues from 45 day old plants. Isoforms ALN1 and ALN2 are consistently expressed at higher levels than ALN3 and ALN4 in all samples. Similar levels of ALN4 are found only in nodules
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
additional information
-
the enzyme has a predicted 22 amino acid signal peptide for the secretory pathway
-
Manually annotated by BRENDA team
additional information
-
the enzyme has a predicted signal peptide for the secretory pathway
-
Manually annotated by BRENDA team
additional information
Q6S4R9, -
the enzyme has a predicted signal peptide for the secretory pathway
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Bacillus halodurans (strain ATCC BAA-125 / DSM 18197 / FERM 7344 / JCM 9153 / C-125)
Escherichia coli (strain K12)
Pseudomonas fluorescens (strain Pf0-1)
Pseudomonas fluorescens (strain Pf0-1)
Pseudomonas fluorescens (strain Pf0-1)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
42000
-
-
IsoII, native PAGE with acrylamide gradient; IsoII, SDS-PAGE, after purification two peaks of 45000 and 42000 Da length showing allantoinase activity are obtained
44000
-
-
gel filtration; SDS-PAGE and gel-filtration
45000
-
-
IsoI, SDS-PAGE, after purification two peaks of 45000 and 42000 Da length showing allantoinase activity are obtained
50000
-
-
gel filtration
53000
-
-
processed recombinant and native enzyme, gel filtration
53000
-
-
IsoI, native PAGE with acrylamide gradient
53300
-
-
calculated from cDNA
57150
-
C6EKV4
gel filtration
93600
-
C6EKV4
gel filtration
125000
-
-
gel filtration
140000
150000
-
gel filtration
140000
-
B5L363, -
gel filtration, estimated molecular mass
189500
-
C6EKV4
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
dimer
-
1 * 54000 + 1 * 48000, SDS-PAGE, the dimer consists of allantoinase the larger subunit and allantoicase
dimer
-
-
homotetramer
B5L363, -
-
homotetramer
P77671
-
monomer
-
1 * 44000, SDS-PAGE; 1 * 44000, SDS-PAGE, gel-filtration
monomer
-
1 * 53000, processed recombinant and native enzyme, SDS-PAGE
tetramer
-
4 * 38000, SDS-PAGE
monomer
-
; 1 * 42000, isozyme IsoII, SDS-PAGE, 1 * 45000, isozyme IsoI, SDS-PAGE
additional information
C6EKV4
gel filtration chromatography reveals a mixture of monomers, dimers, and tetramers
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
proteolytic modification
-
the enzyme has a predicted signal peptide for the secretory pathway
glycoprotein
-
different glycosylation stages as well as signal peptide cleavage lead to different subunits sizes of 53 kDa and 56 kDa
proteolytic modification
-
the enzyme has a predicted 22 amino acid signal peptide for the secretory pathway
proteolytic modification
Q6S4R9, -
the enzyme has a predicted signal peptide for the secretory pathway
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30
-
-
no loss in activity over 6 h
50
-
-
rapidly inactivitad above 50C
50
-
C6EKV4
30 min, comnplete loss of activity
60
70
-
the activity of both protein variants IsoI and IsoII is very resistant to heat treatment because the enzyme is fully active after treatment for 30 min at 60C and shows more than 50% activity after 30 min at 70C
68
-
-
7 min, 78% in activity retained
70
-
-
stable for 10 min
70
-
-
-
90
-
-
60 min, 71% of activity at 30C
ORGANIC SOLVENT
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
deoxycholate
-
20% increase of activity in the presence of 0.1% (w/v) deoxycholate
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
8C, 3 weeks, 90% of activity retains
-
frozen state, 2 weeks
-
4C, 40 mM phosphate buffer, pH 7, 0.01% sodium azide, 1 mM beta-mercaptoethanol, 4 months
-
-20C, 50 mM potassium phosphate buffer, pH 7.5, 15% glycerol, 5 weeks
-
0-5C, 2 weeks, little loss in activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant by gel filtration and affinity chromatography to near homogeneity
-
recombinant enzyme
-
; native enzyme from seeds
-
DEAE-Sephacel column chromatography, MonoQ HR column chromatography, and Sephacryl S-300 gel filtration; native isozymes 1 and 2 from developing fruits 150fold and 286fold, respectively, to homogeneity involving heat treatment at 55C for 20 min, anion exchange chromatography, gel filtration, and another step of anion exchange chromatography
-
Sephacryl S-200 column gel filtration chromatography at 0-4C
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
gene aln, transient expression in Nicotiana tabacum in the endoplasmic reticulum, occasionally also in the Golgi apparatus or the peroxisomes
-
gene AtALN, located on chromosome 4, DNA and amino acid sequence determination and analysis, phylogenetic analysis, functional complementation of an enzyme-deficient yeast dal1 mutant
-
cloned from a hindgut and Malpighian tubule library, DNA and amino acid sequence determination and analysis, sequence comparisons
-
overproduction of enzyme in Escherichia coli
-
expression in Escherichia coli strain BL21
B5L363, -
expression in Saccharomyces cerevisiae; expression in Spodoptera frugiperda
-
gene RpALN, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression analysis, functional complementation of an enzyme-deficient yeast dal1 mutant
Q6S4R9, -
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
the genes involved in the conversion of allantoin into allantoate and in the deamination of guanine to xanthine are organized in three transcriptional units, hpxSAB, hpxC, and guaD. hpxSAB, which includes hpxB encoding allantoinase, is the most tightly regulated unit. This operon is activated by growth on allantoin as a nitrogen source. Addition of allantoin to nitrogen excess cultures does not result in hpxSAB induction. Full induction of hpxSAB by allantoin requires both HpxS, encoding a regulatory protein of the GntR family, and nitrogen assimilation control protein NAC
-
at the onset of flowering, allantoinase gene expression and enzyme increase in shoots and leaves from plants using symbiotically fixed nitrogen as the sole nitrogen source activity
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
D315A
C6EKV4
complete loss of activity
D315N
P77671
no activity
H186A
C6EKV4
complete loss of activity
H242A
C6EKV4
complete loss of activity
H59A
C6EKV4
complete loss of activity
H61A
C6EKV4
complete loss of activity
K146A
C6EKV4
complete loss of activity
N94D
P77671
no activity
N94V
P77671
no activity
R215K
P77671
no significant effect on enzyme activity
S288D
P77671
no significant effect on enzyme activity
S288V
P77671
no significant effect on enzyme activity
S317D
P77671
no activity
S317V
P77671
no activity
additional information
-
construction of an ALN insertion mutant
additional information
-
a T-DNA insertion allantoinase mutant is unable to grow on 10 mM allantoin as the sole nitrogen source, although it still shows some root elongation with a few plants reaching the four-leaf stage, the mutant is rescued by a C-terminally tagged Arabidopsis thaliana AtAln variant, overview
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
purified recombinant ALLase exhibits no activity but can be activated when preincubating with some metal ions before analyzing its activity. Renaturation follows in decreasing order Mn2+, Co2+, Zn2+, Ni2+, Cd2+, Mg2+
C6EKV4