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
-
UniProt
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-glucopyranoside + H2O
4-nitrophenol + beta-D-glucose
show the reaction diagram
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-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 + beta-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
no metal ions are required for the activity of isoform EGCrP1
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 30C 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 30C
-
0.3
D-erythro-dihydrosphingosine
-
-
0.04
D-erythro-sphingosine
-
-
0.149
daidzein-7-glucopyranoside
0.065 - 0.51
galactosylceramide
0.24
galactosylsphingosine
-
-
0.085
genistein-7-glucopyranoside
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, 37C
0.046
quercetin-3-glucopyranoside
0.044
quercetin-4'-glucoside
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 30C
-
2.1
daidzein-7-beta-D-glucopyranoside
6.5
genistein-7-beta-D-glucopyranoside
33.8
lactose
Ovis aries
-
pH 6, 37C
0.5
phlorizin
1
pyridoxine-5'-beta-D-glucoside
Rattus norvegicus
-
pH 6.0, 37C
6.3
quercetin-3-beta-D-glucopyranoside
7.5
quercetin-4'-glucoside
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 30C
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, 37C
0.076
deoxygalactonojirimycin
-
competitive inhibition
0.142
deoxynojirimycin
-
competitive inhibition
56
lactose
-
pH 6.0, 37C
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 37C
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 15C, about 65% activity at 20C, 100% activity at 30-35C, about 30% activity at 40C
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 37C, 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
-20C, 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 LPHCFP, SIYFP 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 20C. 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