Information on EC 3.2.1.62 - glycosylceramidase

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

EC NUMBER
COMMENTARY hide
3.2.1.62
-
RECOMMENDED NAME
GeneOntology No.
glycosylceramidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
a glycosyl-N-acylsphingosine + H2O = a ceramide + a sugar
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of O-glycosyl bond
SYSTEMATIC NAME
IUBMB Comments
glycosyl-N-acylsphingosine glycohydrolase
Broad specificity [cf. EC 3.2.1.45 (glucosylceramidase) and EC 3.2.1.46 (galactosylceramidase)]. Also hydrolyses phlorizin to phloretin and glucose. The intestinal enzyme is a complex that also catalyses the reaction of EC 3.2.1.108 lactase.
CAS REGISTRY NUMBER
COMMENTARY hide
9033-10-7
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
monkey
-
-
Manually annotated by BRENDA team
Ovis aries aries
sheep
-
-
Manually annotated by BRENDA team
strain M-777
-
-
Manually annotated by BRENDA team
strain M-777
-
-
Manually annotated by BRENDA team
puffer fish, no activity in Mycetozoa, slime mold
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
physiological function
-
The profragment, LPHalpha, comprises homologous domains I and II and functions as an intramolecular chaperone in the context of the brush-border LPHbeta region of LPH
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(2-hydroxymethylphenyl)-beta-D-glucopyranoside + H2O
2-hydroxymethylphenol + beta-D-glucose
show the reaction diagram
2-naphthyl-beta-D-galactopyranoside + H2O
naphthol + beta-D-galactopyranose
show the reaction diagram
-
very low activity
-
?
2-nitrophenyl beta-D-galactopyranoside + H2O
2-nitrophenol + beta-D-galactose
show the reaction diagram
2-nitrophenyl beta-D-glucopyranoside + H2O
2-nitrophenol + beta-D-glucose
show the reaction diagram
-
-
-
?
3'-fucosyllactose + H2O
beta-D-glucose + 3-O-(6-deoxy-alpha-L-galactopyranosyl)-beta-D-galactopyranose
show the reaction diagram
-
only oligosaccharides containing galactose as the glycone in beta-linkage hydrolysed
-
?
4'-deoxyphlorizin + H2O
4'-deoxyphloritin + beta-D-glucose
show the reaction diagram
-
low activity
-
?
4-methylumbelliferyl-alpha-L-arabinoside + H2O
4-methylumbelliferone + alpha-L-arabinose
show the reaction diagram
-
-
-
?
4-methylumbelliferyl-beta-D-galactopyranoside + H2O
4-methylumbelliferone + D-galactose
show the reaction diagram
-
-
-
-
?
4-methylumbelliferyl-beta-D-galactoside + H2O
4-methylumbelliferone + beta-D-galactose
show the reaction diagram
4-methylumbelliferyl-beta-D-glucopyranoside + H2O
4-methylumbelliferone + D-glucose
show the reaction diagram
-
-
-
-
?
4-methylumbelliferyl-beta-D-glucoside + H2O
4-methylumbelliferone + beta-D-glucose
show the reaction diagram
-
-
-
?
4-methylumbelliferyl-beta-D-xyloside + H2O
4-methylumbelliferone + beta-D-xylose
show the reaction diagram
-
low activity
-
?
4-nitrophenyl beta-D-fucoside + H2O
4-nitrophenol + beta-D-fucose
show the reaction diagram
-
-
-
?
4-nitrophenyl beta-D-galactopyranoside + H2O
4-nitrophenol + beta-D-galactose
show the reaction diagram
4-nitrophenyl beta-D-glucopyranoside + H2O
4-nitrophenol + beta-D-glucose
show the reaction diagram
4-O-beta-D-galactopyranosyl-D-fructoside + H2O
beta-D-galactose + D-fructose
show the reaction diagram
-
very low activity
-
?
4-O-beta-D-galactopyranosyl-D-glucopyranoside + H2O
D-galactose + beta-D-glucose
show the reaction diagram
4-O-beta-D-galactopyranosyl-D-N-acetylglucosamine + H2O
beta-D-galactose + N-acetylglucosamine
show the reaction diagram
-
poor activity
-
?
4-O-beta-D-glucopyranosyl-D-glucose + H2O
beta-D-glucose + D-glucose
show the reaction diagram
6-bromonaphthyl-beta-D-galactopyranoside
6-bromonaphthol + beta-D-galactopyranose
show the reaction diagram
-
very low activity
-
?
6-O-beta-D-glucopyranosyl-D-glucose + H2O
beta-D-glucose + D-glucose
show the reaction diagram
beta-methylglucoside + H2O
methanol + beta-D-glucose
show the reaction diagram
-
-
-
?
C18-galactosylceramide + H2O
C18-ceramide + D-galactose
show the reaction diagram
-
-
-
-
?
C18-glucosylceramide + H2O
C18-ceramide + D-glucose
show the reaction diagram
-
-
-
-
?
C6-4-nitrobenzo-2-oxa-1,3-diazole-galactosylceramide + H2O
C6-4-nitrobenzo-2-oxa-1,3-diazole-ceramide + D-galactose
show the reaction diagram
C6-4-nitrobenzo-2-oxa-1,3-diazole-glucosylceramide + H2O
C6-4-nitrobenzo-2-oxa-1,3-diazole-ceramide + D-glucose
show the reaction diagram
C6-7-nitro-2,1,3-benzoxadiazole-glucosylceramide + H2O
?
show the reaction diagram
cellotetraose + H2O
? + beta-D-glucose
show the reaction diagram
cellotriose + H2O
? + beta-D-glucose
show the reaction diagram
cellulose + H2O
? + beta-D-glucose
show the reaction diagram
D-erythro-dihydrosphingosine + a sugar
?
show the reaction diagram
D-erythro-sphingosine + a sugar
?
show the reaction diagram
daidzein-7-glucopyranoside + H2O
daidzein + beta-D-glucose
show the reaction diagram
Ovis aries aries
-
-
-
?
daidzin + H2O
3-(4-hydroxyphenyl)-7-hydroxy-chromen-4-one + D-glucose
show the reaction diagram
-
-
-
-
?
daidzin + H2O
?
show the reaction diagram
-
-
-
-
?
galactosyl-N-palmitoylceramide + H2O
beta-D-galactose + N-palmitoylceramide
show the reaction diagram
-
-
-
?
galactosyl-N-palmitoyldihydroceramide + H2O
beta-D-galactose + N-palmitoyldihydroceramide
show the reaction diagram
-
-
-
?
galactosylceramide + H2O
beta-D-galactose + ceramide
show the reaction diagram
-
-
-
?, ir
galactosylceramide + H2O
ceramide + D-galactose
show the reaction diagram
-
-
-
-
?
galactosylsphingosine + H2O
beta-D-galactose + sphingosine
show the reaction diagram
-
very low activity
-
?
galactosylsphingosine + H2O
sphingosine + D-galactose
show the reaction diagram
-
-
-
-
?
genistein-7-glucopyranoside + H2O
genistein + beta-D-glucose
show the reaction diagram
Ovis aries aries
-
-
-
?
genistein-7-glucoside + H2O
genistein + D-glucose
show the reaction diagram
-
-
-
-
?
glucosyl-N-lignoceroyldihydroceramide + H2O
beta-D-glucose + N-lignoceroyldihydroceramide
show the reaction diagram
-
-
-
?
glucosyl-N-palmitoylceramide + H2O
beta-D-glucose + N-palmitoylceramide
show the reaction diagram
-
-
-
?
glucosyl-N-palmitoyldihydroceramide + H2O
beta-D-glucose + N-palmitoyldihydroceramide
show the reaction diagram
-
-
-
?
glucosyl-N-stearoyldihydroceramide + H2O
beta-D-glucose + N-stearoyldihydroceramide
show the reaction diagram
-
-
-
?
glucosylceramide + H2O
?
show the reaction diagram
glucosylceramide + H2O
beta-D-glucose + ceramide
show the reaction diagram
-
-
-
?
glucosylceramide + H2O
ceramide + D-glucose
show the reaction diagram
-
no activity with alpha-galactosylceramide, lactosylceramide, ganglioside M1a and sulfatides
-
-
?
glucosylsphingosine + H2O
beta-D-glucose + N-acylsphingosine
show the reaction diagram
-
very low activity
-
?
glucosylsphingosine + H2O
sphingosine + D-glucose
show the reaction diagram
-
-
-
-
?
glycosyl-N-acylsphingosine + H2O
N-acylsphingosine + a sugar
show the reaction diagram
lacto-N-tetraose + H2O
beta-D-glucose + ?
show the reaction diagram
-
-
-
?
lactose + H2O
?
show the reaction diagram
-
-
-
-
?
lactose + H2O
D-galactose + glucose
show the reaction diagram
-
reaction is not inhibited by 2-fluoro-dinitrophenylglucoside
-
-
?
lactose + H2O
D-glucose + D-galactose
show the reaction diagram
lactosyl-N-lignoceroyldihydroceramide + H2O
beta-D-glucose + D-galactose + N-lignoceroyldihydroceramide
show the reaction diagram
-
-
-
?
lactosyl-N-palmitoyldihydroceramide + H2O
beta-D-glucose + D-galactose + N-palmitoyldihydroceramide
show the reaction diagram
-
-
-
?
lactosyl-N-stearoyldihydroceramide + H2O
beta-glucose + D-galactose + N-stearoyldihydroceramide
show the reaction diagram
-
-
-
?
lactosylceramide + H2O
beta-D-glucose + galactosylceramide
show the reaction diagram
-
low activity
-
?
N-palmitoyldihydroglucosylceramide + H2O
beta-D-glucose + ?
show the reaction diagram
-
-
-
?
N-palmitoyldihydroglucosylcerebroside + H2O
beta-D-glucose + N-palmitoyldihydrocerebroside
show the reaction diagram
-
1.5% of the activity towards lactose
-
?
oligoglycosylglucosyl-(1->1)-ceramide + H2O
ceramide + oligoglycosylglucose
show the reaction diagram
p-nitrophenyl-beta-cellobioside + H2O
p-nitrophenyl-beta-D-glucoside + D-glucose
show the reaction diagram
-
-
-
-
?
p-nitrophenyl-beta-lactoside + H2O
p-nitrophenyl-beta-D-glucoside + D-galactose
show the reaction diagram
-
-
-
-
?
p-nitrophenyl-beta-maltoside + H2O
p-nitrophenol-beta-D-glucoside + D-glucose
show the reaction diagram
-
-
-
-
?
phenyl-beta-D-galactopyranoside + H2O
phenol + beta-D-galactopyranose
show the reaction diagram
-
low activity
-
?
phloretin-beta-D-galactoside + H2O
phloretin + beta-D-galactose
show the reaction diagram
-
very low activity
-
?
phlorizin + H2O
?
show the reaction diagram
phlorizin + H2O
phloretin + beta-D-glucose
show the reaction diagram
pyridoxine-5'-beta-D-glucoside + H2O
pyridoxine + D-glucose
show the reaction diagram
-
-
-
-
r
quercetin 3-O-beta-D-glucopyranoside + H2O
quercetin + beta-D-glucose
show the reaction diagram
Ovis aries aries
-
-
-
?
quercetin 4'-O-beta-D-glucopyranoside + H2O
quercetin + beta-D-glucose
show the reaction diagram
Ovis aries aries
-
flavone glucosides more efficiently hydrolysed than isoflavone glucosides, major contribution to the hydrolysis is from the lactase site
-
?
quercetin-3-glucoside + H2O
quercetin + D-glucose
show the reaction diagram
-
reaction is marginally inhibited by 2-fluoro-dinitrophenylglucoside
-
-
?
quercetin-4'-glucoside + H2O
quercetin + D-glucose
show the reaction diagram
-
reaction is marginally inhibited by 2-fluoro-dinitrophenylglucoside
-
-
?
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
(2-hydroxymethylphenyl)-beta-D-glucopyranoside + H2O
2-hydroxymethylphenol + beta-D-glucose
show the reaction diagram
3'-fucosyllactose + H2O
beta-D-glucose + 3-O-(6-deoxy-alpha-L-galactopyranosyl)-beta-D-galactopyranose
show the reaction diagram
-
only oligosaccharides containing galactose as the glycone in beta-linkage hydrolysed
-
?
4-O-beta-D-galactopyranosyl-D-fructoside + H2O
beta-D-galactose + D-fructose
show the reaction diagram
-
very low activity
-
?
4-O-beta-D-galactopyranosyl-D-glucopyranoside + H2O
D-galactose + beta-D-glucose
show the reaction diagram
4-O-beta-D-galactopyranosyl-D-N-acetylglucosamine + H2O
beta-D-galactose + N-acetylglucosamine
show the reaction diagram
-
poor activity
-
?
4-O-beta-D-glucopyranosyl-D-glucose + H2O
beta-D-glucose + D-glucose
show the reaction diagram
6-O-beta-D-glucopyranosyl-D-glucose + H2O
beta-D-glucose + D-glucose
show the reaction diagram
cellotetraose + H2O
? + beta-D-glucose
show the reaction diagram
cellotriose + H2O
? + beta-D-glucose
show the reaction diagram
cellulose + H2O
? + beta-D-glucose
show the reaction diagram
galactosylceramide + H2O
beta-D-galactose + ceramide
show the reaction diagram
-
-
-
ir
galactosylsphingosine + H2O
beta-D-galactose + sphingosine
show the reaction diagram
-
very low activity
-
?
glucosylceramide + H2O
beta-D-glucose + ceramide
show the reaction diagram
-
-
-
?
glucosylsphingosine + H2O
beta-D-glucose + N-acylsphingosine
show the reaction diagram
-
very low activity
-
?
lacto-N-tetraose + H2O
beta-D-glucose + ?
show the reaction diagram
-
-
-
?
lactosylceramide + H2O
beta-D-glucose + galactosylceramide
show the reaction diagram
-
low activity
-
?
N-palmitoyldihydroglucosylceramide + H2O
beta-D-glucose + ?
show the reaction diagram
-
-
-
?
oligoglycosylglucosyl-(1->1)-ceramide + H2O
ceramide + oligoglycosylglucose
show the reaction diagram
phlorizin + H2O
?
show the reaction diagram
-
-
-
-
?
phlorizin + H2O
phloretin + beta-D-glucose
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2',4'-dinitrophenyl-2-deoxy-2-fluoro-beta-D-galactopyranoside
-
inactivation of lactase hydrolase activity
2',4'-dinitrophenyl-2-deoxy-2-fluoro-beta-D-glucopyranoside
-
inactivation of phlorizin hydrolase activity
2',4'-dinitrophenyl-2-fluoro-2-deoxy-beta-D-glucopyranoside
Ovis aries aries
-
rapidly inhibits phlorizin hydrolase activity by covalently binding to the glutamic acid residue of the active site, lactase activity more slowly inhibited, pre-incubation with glucal protects the lactase active site from inactivation
2-fluoro-dinitrophenylglucoside
-
specifically inhibits phlorizin hydrolase, lactase activity is inhibited slowly
2-oxoglutaric acid
-
50% inhibition of the tartaric acid-activated phlorizin hydrolase at 20 mM
Ca2+
about 78% residual activity at 1 mM
cellobiose
cerebroside
-
from brain, competitive inhibition of lactase activity, Ki: 1.11 mM
Co2+
about 18% residual activity at 1 mM
conduritol-B-epoxide
-
90% inactivation of lactase hydrolysis, 55% inactivation of phlorizin hydrolysis, inactivation by covalent binding to Glu1747 and Glu 1271
Cu2+
about 10% residual activity at 1 mM
D-Galactal
-
inactivation of lactase hydrolase activity
D-Galactono-1,4-lactone
D-glucal
-
inactivation of lactase hydrolase activity
D-glucono-1,5-lactone
D-glucose
-
-
deoxygalactonojirimycin
-
competitive inhibition, Ki: 0.076 mM
deoxynojirimycin
-
competitive inhibition, Ki: 0.142 mM
DMSO
the enzyme is inhibited by 30-50% (v/v) DMSO
epigallocatechin-3-gallate
-
inhibits in vitro hydrolysis of lactose by lactase phlorizin hydrolase
Fe2+
about 40% residual activity at 1 mM
Fe3+
about 50% residual activity at 1 mM
galactosylceramide
-
competitive inhibition of glucosyldihydroceramide hydrolysis
glucosyldihydroceramide
-
competitive inhibition of galactosylceramide hydrolysis
Hg+
complete inhibition at 1 mM
lactose
-
competitive to pyridoxine-5'-beta-D-glucoside
Maleic acid
-
55% inhibition for the tartaric acid-stimulated phlorizin hydrolase at 20 mM
Mg2+
about 90% residual activity at 1 mM
Mn2+
about 50% residual activity at 1 mM
N-(n-butyl)-deoxygalactojirimycin
-
-
n-butyldeoxygalactonojirimycin
-
-
Na+
about 93% residual activity at 1 mM
NaCl
-
30% inhibition of the tartaric acid-stimulated phlorizin hydrolase at 20 mM
Ni2+
about 5% residual activity at 1 mM
NSP4
-
secreted non-structural rotavirus protein
-
oxalacetic acid
-
50% inhibition of the tartaric acid-stimualted phlorizin hydrolase at 20 mM
oxalic acid
-
47% inhibition of the tartaric acid-activated phlorizin hydrolase at 20 mM
Pb2+
about 10% residual activity at 1 mM
phlorizin
pyridoxine-5'-beta-D-glucoside
-
substrate inhibition above 3 mM
Pyruvic acid
-
strongly inhibitory for the tartaric acid-stimulated phlorizin hydrolase
SO42-
-
strong inhibition of the tartaric acid-activated phlorizin hydrolase at 20 mM
sodium cholate
-
competitive inhibition of lactase activity, in the presence of bovine serum albumin
Tris(hydroxymethyl)aminomethane
Triton X-100
strongly inhibits the activity even at a low concentration
Zn2+
about 8% residual activity at 1 mM
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cis-aconitic acid
-
more than 2fold stimulation only for phlorizin activity
Citric acid
-
highest stimulation at 50 mM and 30°C only for phlorizin activity
EDTA
about 120% activity at 1 mM
isocitric acid
-
1.7fold stimulation only for phlorizin activity
L-ascorbic acid
-
1.6fold stimulation only for phlorizin activity
L-Tartaric acid
-
most stimulating effect only for phlorizin activity
Pyruvic acid
-
0.5fold stimulation only for phlorizin activity
sodium cholate
-
increases the solubility of the substrates
taurodeoxycholate
-
activation
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4.64
(6-[N-(7-nitrobenz-2-oxa-1,3 diazol-4-yl)amino]hexanoyl)-beta-D-glucosylceramide
-
-
13.1
2-nitrophenyl-beta-D-galactopyranoside
-
-
0.45
4-methylumbelliferyl-alpha-L-arabinoside
-
-
144.5
4-methylumbelliferyl-beta-D-galactopyranoside
-
-
0.24
4-methylumbelliferyl-beta-D-galactoside
-
-
49.28
4-methylumbelliferyl-beta-D-glucopyranoside
-
-
0.53
4-methylumbelliferyl-beta-D-glucoside
-
-
1.4
4-methylumbelliferyl-beta-D-xyloside
-
-
0.36
4-nitrophenyl-beta-D-fucoside
-
-
22.3
4-nitrophenyl-beta-D-galactopyranoside
-
-
4.1
4-nitrophenyl-beta-D-glucopyranoside
-
-
67
4-O-beta-D-galactopyranosyl-D-fructoside
-
-
5.1 - 41.7
4-O-beta-D-galactopyranosyl-D-glucopyranoside
4.8
4-O-beta-D-galactopyranosyl-D-glucosamine
-
-
1.1 - 2.78
4-O-beta-D-glucopyranosyl-D-glucopyranoside
4.4
4-O-beta-D-glucopyranosyl-D-glucospyranoside
-
-
-
22
6-O-beta-D-glucopyranosyl-D-glucopyranoside
-
-
9.24
C18-galactosylceramide
-
-
13.67
C18-glucosylceramide
-
-
2.04
C6-4-nitrobenzo-2-oxa-1,3-diazole-galctosylceramide
-
-
0.0058
C6-7-nitro-2,1,3-benzoxadiazole-glucosylceramide
at pH 7.3 and 30°C
-
0.3
D-erythro-dihydrosphingosine
-
-
0.04
D-erythro-sphingosine
-
-
0.149
daidzein-7-glucopyranoside
Ovis aries aries
-
-
0.149
daidzein-7-glucoside
-
pH 6, 37°C
0.065 - 0.51
galactosylceramide
0.24
galactosylsphingosine
-
-
0.085
genistein-7-glucopyranoside
Ovis aries aries
-
-
0.085
genistein-7-glucoside
-
pH 6, 37°C
0.01
glucosylceramide
-
-
0.62
glucosyldihydroceramide
-
-
0.37
glucosylsphingosine
-
-
8.8 - 30
lactose
0.27
lactosylceramide
-
-
0.73 - 1.12
N-palmitoyldihydrocerebroside
0.542 - 18.13
p-nitrophenyl-beta-cellobioside
1.67 - 21.62
p-nitrophenyl-beta-lactoside
2.82 - 5.12
p-nitrophenyl-beta-maltoside
0.4
phloretin-beta-D-galactoside
-
-
0.002 - 0.44
phlorizin
1
pyridoxine-5'-beta-D-glucoside
-
pH 6.0, 37°C
0.046
quercetin-3-glucopyranoside
Ovis aries aries
-
-
0.046
quercetin-3-glucoside
-
pH 6, 37°C
0.044
quercetin-4'-glucoside
-
pH 6, 37°C
0.044
quercetin-4-glucopyranoside
Ovis aries aries
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
33.8
4-O-beta-D-galactopyranosyl-D-glucopyranoside
Ovis aries aries
-
-
0.0383
C6-7-nitro-2,1,3-benzoxadiazole-glucosylceramide
Cryptococcus neoformans
H1AE12
at pH 7.3 and 30°C
-
2.1
daidzein-7-beta-D-glucopyranoside
Ovis aries aries
-
-
2.1
daidzein-7-glucoside
Ovis aries
-
pH 6, 37°C
6.5
genistein-7-beta-D-glucopyranoside
Ovis aries aries
-
-
6.5
genistein-7-glucoside
Ovis aries
-
pH 6, 37°C
33.8
lactose
Ovis aries
-
pH 6, 37°C
0.5
phlorizin
1
pyridoxine-5'-beta-D-glucoside
Rattus norvegicus
-
pH 6.0, 37°C
6.3
quercetin-3-beta-D-glucopyranoside
Ovis aries aries
-
-
6.3
quercetin-3-glucoside
Ovis aries
-
pH 6, 37°C
7.5
quercetin-4'-glucoside
Ovis aries
-
pH 6, 37°C
7.5
quercetin-4-beta-D-glucopyranoside
Ovis aries aries
-
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
6.6
C6-7-nitro-2,1,3-benzoxadiazole-glucosylceramide
Cryptococcus neoformans
H1AE12
at pH 7.3 and 30°C
202165
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.11
cerebroside
-
from brain, competitive inhibition of lactase activity
64
D-glucose
-
pH 6.0, 37°C
0.076
deoxygalactonojirimycin
-
competitive inhibition
0.142
deoxynojirimycin
-
competitive inhibition
56
lactose
-
pH 6.0, 37°C
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.074
epigallocatechin-3-gallate
Rattus norvegicus
-
in 1.5 mM sodium phosphate buffer (pH 6.8), at 37°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00016
-
activity in CHOP cell lysates transformed with cDNA encoding KLPH, with conduritol B epoxide, C6-4-nitrobenzo-2-oxa-1,3-diazole-galactosylceramide as substrate
0.00025
-
activity in CHOP cell lysates transformed with cDNA encoding KLPH, without conduritol B epoxide, C6-4-nitrobenzo-2-oxa-1,3-diazole-galactosylceramide as substrate
0.0005
0.00133
-
in cytosolic fraction with conduritol B epoxide, C6-4-nitrobenzo-2-oxa-1,3-diazole-glucosylceramide as substrate
0.00164
-
in non infected cells, cell homogenates
0.00167
-
in cells infected with rhesus monkey rotavirus, cell homogenates
0.001875
-
activity in CHOP cell lysates transformed with cDNA encoding KlrP, with conduritol B epoxide, C6-4-nitrobenzo-2-oxa-1,3-diazole-glucosylceramide as substrate
0.00219
-
in cells infected with rhesus monkey rotavirus, in brush border-membrane enriched fraction
0.0025
-
activity in CHOP cell lysates transformed with cDNA encoding KLPH, without conduritol B epoxide, C6-4-nitrobenzo-2-oxa-1,3-diazole-glucosylceramide as substrate
0.0028
-
in cells infected with rhesus monkey rotavirus and NSP4 neutralizing antibodies, in brush border-membrane enriched fraction
0.00334
-
in non infected cells, in brush border-membrane enriched fraction
0.00359
-
in cells infected with inactivated rhesus monkey rotavirus, in brush border-membrane enriched fraction
0.00375
-
activity in CHOP cell lysates transformed with cDNA encoding KlrP, without conduritol B epoxide, C6-4-nitrobenzo-2-oxa-1,3-diazole-glucosylceramide as substrate
0.0052
-
activity in CHOP cell lysates transformed with cDNA encoding KlrP, with conduritol B epoxide, C6-4-nitrobenzo-2-oxa-1,3-diazole-galactosylceramide as substrate
0.0066
-
activity in CHOP cell lysates transformed with cDNA encoding KlrP, without conduritol B epoxide, C6-4-nitrobenzo-2-oxa-1,3-diazole-galactosylceramide as substrate
0.45
-
phlorizin activity
1.78
-
galactosylceramide hydrolysis
2.67
-
glucosylceramide hydrolysis
5 - 6
-
lactase activity
15.33
-
lactose hydrolysis
16.1
-
lactase activity
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.2 - 5.6
-
phlorizin activity in acetate buffer
5.2
-
hydrolysis of galactosyl- or glucosylceramide in the presence of taurodeoxycholate
5.5
-
-
5.5 - 5.7
-
hydrolysis of disaccharides or artificial substrates
5.6 - 6.2
-
-
5.8 - 6
-
-
6 - 7
-
C6-4-nitrobenzo-2-oxa-1,3-diazole-glucosylceramide as substrate
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 8
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15 - 40
about 50% activity at 15°C, about 65% activity at 20°C, 100% activity at 30-35°C, about 30% activity at 40°C
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
transfected Cos-1 cells
Manually annotated by BRENDA team
additional information
-
sequential passage of LPH through Rab4-, Rab8- and Rab11-containing compartments. Trafficking of LPH from the TGN to the cell surface can be modulated by dominant-negative Rab8
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
38000
-
sequence analysis
57000
-
SDS-PAGE
130000
-
mature form, proteolytically cleaved from precursor form, SDS-PAGE
135000
-
SDS-PAGE
145000
-
SDS-PAGE, mature form
160000
215000
220000
230000
-
mannose-rich form, SDS-PAGE
245000
280000
-
gel filtration, hydrophilic form
290000 - 300000
-
gel filtration
320000
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 135000-140000, SDS-PAGE, mature protein
monomer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
proteolytic modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
of the recombinant protein with the hanging drops by vapor diffusion method
-
by vapor diffusion in the presence of detergents like Triton X-100, octyl beta-D-glucopyranoside, decyl beta-D-maltopyranoside, and dodecyl beta-D-maltopyranoside, form, in complex with the ganglioside GM3
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.2
-
little activity below
208757
4.2 - 9
-
little activity outside this range
208756
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
no detectable loss of activity for several weeks
46
-
34-70% remaining activity after 30 min, depending on substrates
49
-
lactase activity faster destroyed than phlorizin activity
55
-
complete inactivation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
at neutral pH and 37°C, lactase acitivity rapidly lost, partially prevented by p-chloromercuribenzoate, more stable if attached to its border-membrane
-
sensitive to repeated freezing and thawing
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, several months
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
antibody affinity chromatography
-
by magnesium precipitation method
-
GSTrap column chromatography
Helix pomatia lectin chromatography
-
highly purified
-
of the recombinant protein by a hi-trap chelating HP column and superdex 200 column chromatography
-
of the recombinant protein by by Ni(II) affinity chromatography
-
to homogeneity
Ovis aries aries
-
to homogeneity, chromatography steps
-
to homogeneity, chromatography techniques
to homogeneity, immunoadsorbent chromatography
-
to homogeneity, immunoadsorbent chromatography, amphiphilic form
-
to homogeneity, immunoaffinity chromatography
-
two isoenzymes from proximal and distal intestine, to homogeneity, chromatography techniques
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
COS-1 cells triple transfected with LPH–CFP, SI–YFP and Rab4, Rab8 or Rab11 fused to DsRed. MDCK cells stably expressing LPHmyc
-
expressed in CACO-2 cells; LPH 5' upstream subcloned from rat liver genomic DNA
-
expressed in Escherichia coli BL21(DE3) cells
expression in Chinese hamster ovary-derived cells expressing polyoma LT antigen
-
expression in Chinese hamster ovary-derived cells expressing polyoma LT antigen, in HEK293 cells cotransfected with in KlrP siRNA, overexpression in Escherichia coli
-
expression in Cos-1 cells
-
expression of mutants in Cos-7 cells
-
expression of the gene lacking the 30-residue N-terminal secretion signal sequence in Escherichia coli, selenomethionine-derivatized enzyme
-
genotyping
-
mutant G1363S introduced into the wild-type LPH cDNA cloned in the vector pSG5. Mutant transiently expressed in COS-1 cells
-
transient expression of wild-type and domain deletion mutant enzymes and isolated enzyme domains in Cos-1 cells, expression in MDCK cells
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E254Q
-
inactive
-
E483Q
-
inactive
-
D1711N
-
introduction of a glycosylation site, reduced transport rate to the plasma membrane; introduction of potential N-glycosylation site, no enzymic activity, probably due to altered protein folding pattern and reduced dimerization efficiency
E165D
-
reduced activity
E165Q
-
almost no activity
E373D
-
reduced activity
E373Q
-
almost no activity
G1363S
-
the mutant protein is malfolded and enzymatically inactive and can not exit the endoplasmic reticulum. The mutation creates an additional N-glycosylation site that is characteristic of a temperature-sensitive protein. The potential glycosylation site generated by the mutation is not the cause of defective trafficking of LPH-G1363S or its reduced enzymatic activity. Intracellular transport and enzymatic activity, but not correct folding are partially restored by expression at 20°C. The mutant is responsible for an increased turnover rate
G1363S/N1361A
-
eliminates the N-glycosylation site, does not restore the features of wild-type LPH
I1697N
-
introduction of a glycosylation site, reduced transport rate to the plasma membrane; introduction of potential N-glycosylation site, no enzymic activity, probably due to altered protein folding pattern and reduced dimerization efficiency
P1743S
-
introduction of a glycosylation site, reduced transport rate to the plasma membrane; introduction of potential N-glycosylation site, no enzymic activity, probably due to altered protein folding pattern and reduced dimerization efficiency
E233A
-
site directed mutagenesis
E351S
-
site directed mutagenesis
E233A
-
site directed mutagenesis
-
E351S
-
site directed mutagenesis
-
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
additional information
-
regulation of LPH exocytosis by multiple Rab GTPases