Information on EC 2.4.1.210 - limonoid glucosyltransferase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.4.1.210
-
RECOMMENDED NAME
GeneOntology No.
limonoid glucosyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
UDP-glucose + limonin = glucosyl-limonin + UDP
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexosyl group transfer
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
UDP-glucose:limonin glucosyltransferase
The enzyme purified from navel orange albedo tissue also acts on the related tetranortriterpenoid nomilin.
CAS REGISTRY NUMBER
COMMENTARY hide
195836-82-9
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
sweet lime
-
-
Manually annotated by BRENDA team
citron
-
-
Manually annotated by BRENDA team
Osbeck
-
-
Manually annotated by BRENDA team
strawberry
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
the enzyme belongs to clade C of flavonoid and limonoid glucosyltransferases
additional information
-
limonoid glucosyltransferase is an enzyme that catalyzes the conversion of bitter limonoid into non-bitter limonoid glucoside while retaining the health benefit of limonoids in the juice
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
limonin + UDP-glucose
glucosyl-limonin + UDP
show the reaction diagram
nomilin + UDP-glucose
glucosyl-nomilin + UDP
show the reaction diagram
-
-
-
-
?
nomilin + UDP-glucose
UDP + nomilin 17-beta-D-glucopyranoside
show the reaction diagram
UDP-glucose + limonin
glucosyl-limonin + UDP
show the reaction diagram
UDP-glucose + limonin
UDP + limonin 17-beta-D-glucopyranoside
show the reaction diagram
-
-
-
-
?
UDP-glucose + limonoate lactone
UDP + glucosyl-limonoate lactone
show the reaction diagram
-
-
-
?
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
limonin + UDP-glucose
glucosyl-limonin + UDP
show the reaction diagram
nomilin + UDP-glucose
UDP + nomilin 17-beta-D-glucopyranoside
show the reaction diagram
UDP-glucose + limonin
glucosyl-limonin + UDP
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
-
5 mM, 30% stimulation
Zn2+
-
5 mM, 13% stimulation
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-ethyl-3-(3-dimethylaminopropyl) carbodiimide
-
modifies tryptophan residues
3-nitro-L-tyrosine ethylester
-
modifies tryptophan residues
Ca2+
-
10 mM, 13% inhibition
citraconic anhydride
Cu2+
-
5 mM, 58% inhibition
diethyl dicarbonate
-
1 mM, 76% inhibition
diethyldicarbonate
-
modifies histidine residues and inactivates the enzyme, pseudo first order kinetics. When the histidine modification is reversed by hydroxylamine treatment, 79% of the activity is restored
EDTA
-
1 mM, complete inactivation
Hg2+
-
1 mM, 33% inhibition
Mg2+
-
10 mM, 22% inhibition
N-bromosuccinimide
-
modifies tryptophan residues and inactivates the enzyme, loss of LGTase activity by NBS treatment is partially protected by pre-incubating the enzyme with excess limonin
additional information
-
inactivation of the enzyme following modification of carboxyl and imidazole moieties is a consequence of a loss in substrate binding and catalysis in the glucosyltransfer reaction. No inhibition by serine modifiying diisopropyl fluorophosphate and cysteine modifying 4-chloromercuribenzoate and iodoacetamide
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.065 - 0.2
limonin
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
91.7
limonin
Citrus maxima x Citrus paradisi
-
pH 7.0-8.4, 25C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.013
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.4
-
enzyme immobilized covalently on chitosan cross-linked with gluteraldehyde (Chito-GA-LGTase)
8.2
-
enzyme immobilized ionically on DEAE-Toyopearl (DE-Tp-LGTase)
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 9.5
-
activity range
6.5 - 9.5
-
pH 6.5: about 70% of maximal activity, pH 9.5: about 55% of maximal activity
6.5 - 9
-
-
additional information
-
at an acidic pH the D-ring of the substrate that should be in an open form is closed and glycosylation does not occur. The decrease in activity at acidic pH also indicates that an essential residue participates in catalytic functioning while in the unprotonated state
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
assay at room temperature
40
-
enzyme immobilized covalently on chitosan cross-linked with gluteraldehyde (Chito-GA-LGTase) and enzyme immobilized covalently on cellulose carbonate (Ce-Ca-LGTase)
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 60
-
20C: about 40% of maximal activity, 60C: about 40% of maximal activity
20 - 70
-
20C: about 55% of maximal activity, 70C: about 30% of maximal activity
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
suspension
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
56000
-
Newhall navel orange, SDS-PAGE
57500
-
predicted from amino acid sequence
58000
-
Frost navel orange, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
-
56000-58000, SDS-PAGE
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.5 - 9.5
-
4C, 12 h, stable
658786
6.5
-
activity below pH 6.5 is inhibited by a closed D-ring
288690
10.5
-
4C, 12 h, 62% loss of activity
658786
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
-
pH 7.5, 3 h, Ce-Ca-LGTase retains 65.3%, free LGTase retains 19.2%
60
-
pH 7.5, 30 min, free enzyme retains 55% of the activity, enzyme immobilized ionically on DEAE-Toyopearl retains 80% of the activity, enzyme immobilized covalently on cellulose carbonate retains 45% of the activity, enzyme immobilized covalently on chitosan cross-linked with gluteraldehyde retains 40% of the activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
immobilized LGTase retains more than 80% activity up to 15 cycles on cellulose carbonate, nine cycles on chitosan and six cycles on DEAE-Toyopearl
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80C, 10 mM Mes-KOH, pH 7 allows frozen storage with full recovery of activity
-
-80C, 50 mM Tris-HCl, pH 8.0, all acivity is lost
-
4C, pH 7.5, 10 mM Tris-HCl, enzyme retains 95% of the activity after 3 months, enzyme retains 73% of the activity after 9 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme from Escherichia coli and from Pichia pastoris
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cDNA clone encoding limonoid UDP-glucosyltransferase isolated, single copy gene in the citrus genome
-
expression in Escherichia coli for sequencing
gene SLLGT, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression in Escherichia coli and in Pichia pastoris
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
food industry
-
Limonoid glucosyltransferase is an enzyme that catalyzes the conversion of bitter limonoid into non-bitter limonoid glucoside while retaining the health benefit of limonoids in the juice. The immobilization of this enzyme in a column can solve the juice bitterness problem
medicine
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pharmacological application of limonoid GTase gene
nutrition