Information on EC 3.2.1.123 - endoglycosylceramidase

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

EC NUMBER
COMMENTARY hide
3.2.1.123
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RECOMMENDED NAME
GeneOntology No.
endoglycosylceramidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
oligoglycosylglucosyl-(1<->1)-ceramide + H2O = ceramide + oligoglycosylglucose
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis
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EGALC hydrolyzes 6-gala series glycosphingolipids to produce intact oligosaccharides and ceramides
hydrolysis of O-glycosyl bond
-
-
-
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transglycosylation
-
EGALC transfers oligosaccharides from the glycosphingolipids to acceptors such as fluorescent 1-alkanols
SYSTEMATIC NAME
IUBMB Comments
oligoglycosylglucosyl-(1<->1)-ceramide glycohydrolase
An enzyme from Rhodococcus sp. that degrades various acidic and neutral glycosphingolipids to oligosaccharides and ceramides, by cleaving a glucosyl bond. Does not act on monoglycosylceramides. cf. EC 3.2.1.62 glycosylceramidase.
CAS REGISTRY NUMBER
COMMENTARY hide
105503-61-5
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Aiptasia sp.
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
Cellana sp.
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Manually annotated by BRENDA team
Condylactis sp.
seaflower
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Manually annotated by BRENDA team
variant grubii, serotype A
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Manually annotated by BRENDA team
variant grubii, serotype A
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Manually annotated by BRENDA team
Entacmaea sp.
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Manually annotated by BRENDA team
strain 105
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Manually annotated by BRENDA team
strain 105
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Manually annotated by BRENDA team
Marphysa sp.
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Manually annotated by BRENDA team
Meretrix sp.
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Manually annotated by BRENDA team
strain C9
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Manually annotated by BRENDA team
G-74-2
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
Ruditapes sp.
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-N-hexadecanoylamino-4-nitrophenyl-O-beta-lactoside + H2O
?
show the reaction diagram
-
derivative possessing an N-acyl chain, 4-nitrophenyllactoside chromogenic substrate C16ampNPLac
-
-
?
4-methylumbelliferyl beta-D-glucoside + H2O
4-methylumbelliferone + beta-D-glucose
show the reaction diagram
4-nitrophenyl alpha-D-glucoside + H2O
4-nitrophenol + alpha-D-glucose
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-galactoside + H2O
4-nitrophenol + beta-D-galactose
show the reaction diagram
4-nitrophenyl beta-D-glucoside + H2O
4-nitrophenol + beta-D-glucose
show the reaction diagram
4-nitrophenyl beta-D-xyloside + H2O
4-nitrophenol + beta-D-xylose
show the reaction diagram
7-nitro-2,1,3-benzoxadiazole-pentanol + digalactosyldiacylglycerol
?
show the reaction diagram
-
transglycosylation
-
-
?
alpha-L-fucosyl-(1->2)-ganglioside GM1a + H2O
?
show the reaction diagram
-
isoform EGCase I exhibits 100% activity with this substrate
-
-
?
blood group A type 1 heptaglycosylceramide + H2O
?
show the reaction diagram
-
-
-
-
?
blood group A type 2 heptaglycosylceramide + H2O
?
show the reaction diagram
-
-
-
-
?
C6-7-nitro-2,1,3-benzoxadiazole-glucosylceramide + H2O
?
show the reaction diagram
-
-
-
-
?
D-erythro-dihydrosphingosine + H2O
?
show the reaction diagram
D-erythro-sphingosine + H2O
?
show the reaction diagram
Fucalpha(1-3)GalNAcalpha(1-3)(Fucalpha(1-3))Galbeta(1-4)Glcbeta(1-1)ceramide + H2O
Fucalpha(1-3)GalNAcalpha(1-3)(Fucalpha(1-3))Galbeta(1-4)Glc + ceramide
show the reaction diagram
-
84% of the activity with galactosylcerebroside
-
?
Galalpha(1-4)Galbeta(1-4)Glcbeta(1-1)ceramide + H2O
Galalpha(1-4)Galbeta(1-4)Glc + ceramide
show the reaction diagram
-
i.e. Gb3Cer
-
-
?
Galalpha1-6Galalpha1-6Galbeta1-6Galbeta1-1'ceramide + H2O
ceramide + Galalpha1-6Galalpha1-6Gal
show the reaction diagram
-
most rapidly hydrolyzed
-
-
?
Galalpha1-6Galbeta1-3-diacylglycerol + H2O
?
show the reaction diagram
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-
-
-
?
Galalpha1-6Galbeta1-6Galbeta1-1'ceramide + H2O
ceramide + Galalpha1-6Galbeta1-6Gal
show the reaction diagram
-
-
-
-
?
Galbeta(1,3)GalNacbeta(1,4)Galbeta(1,4)Glcbeta-1,1'-ceramide + H2O
ceramide + Galbeta(1,3)GalNacbeta(1,4)Galbeta(1,4)Glc
show the reaction diagram
Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,3)]Galbeta(1,4)Glcbeta-1,1'-ceramide + H2O
ceramide + Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,3)]Galbeta(1,4)Glc
show the reaction diagram
Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,8)NeuAcalpha(2,3)]Galbeta(1,4)Glcbeta-1,1'-ceramide + H2O
ceramide + Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,8)NeuAcalpha(2,3)]Galbeta(1,4)Glc
show the reaction diagram
Galbeta(1,4)Glcbeta-1,1'-ceramide + H2O
ceramide + Galbeta(1,4)Glc
show the reaction diagram
Galbeta(1,6)Galbeta(1,6)Galbeta-1,1'-ceramide + H2O
?
show the reaction diagram
Galbeta(1,6)Galbeta(1,6)Glcbeta-1,1'-ceramide + H2O
ceramide + Galbeta(1,6)Galbeta(1,6)Glc
show the reaction diagram
-
37C, pH 3.0, 0.2% (w/v) Triton X-100
-
-
?
Galbeta(1-3)GalNAcbeta(1-4)(NeuAcalpha(2-3))Galbeta(1-4)Glcbeta(1-1)ceramide + H2O
Galbeta(1-3)GalNAcbeta(1-4)(NeuAcalpha(2-3))Galbeta(1-4)Glc + ceramide
show the reaction diagram
Galbeta(1-3)GalNAcbeta(1-4)(NeuAcalpha(2-3))Galbeta(1-4)Glcbeta(1-1)ceramide + methanol
?
show the reaction diagram
-
i.e. GM1
products may be attributed to GM-1 oligosaccharide II3NeuAcGgOse4 and methyl-GM-1 oligosaccharide methyl-II3NeuAcGgOse4
?
Galbeta(1-3)GalNAcbeta(1-4)(NeuAcalpha(2-3))Galbeta(1-4)Glcbeta(1-1)ceramide 4-nitrobenzo-2-oxy-1,3-diazole + H2O
Galbeta(1-3)GalNAcbeta(1-4)(NeuAcalpha(2-3))Galbeta(1-4)Glc + 4-nitrobenzo-2-oxy-1,3-diazole ceramide
show the reaction diagram
Galbeta(1-3)GalNAcbeta(1-4)(NeuAcalpha(2-8)NeuAcalpha(2-3))Galbeta(1-4)Glcbeta(1-1)ceramide + H2O
Galbeta(1-3)GalNAcbeta(1-4)(NeuAcalpha(2-8)NeuAcalpha(2-3))Galbeta(1-4)Glc + ceramide
show the reaction diagram
Galbeta(1-3)GalNAcbeta(1-4)Galbeta(1-4)Glcbeta(1-1)ceramide + H2O
Galbeta(1-3)GalNAcbeta(1-4)Galbeta(1-4)Glc + ceramide
show the reaction diagram
Galbeta(1-4)Glcbeta(1-1)ceramide + H2O
Galbeta(1-4)Glc + ceramide
show the reaction diagram
Galbeta(1-6)Galbeta(1-6)Galbeta(1-1)ceramide + H2O
Galbeta(1-6)Galbeta(1-6)Gal + ceramide
show the reaction diagram
-
i.e. neogalactosylceramide, 77% of the activity with lactosylceramide
-
?
Galbeta1-1'-ceramide + H2O
?
show the reaction diagram
-
-
-
-
?
Galbeta1-6Galbeta1-1'-ceramide + H2O
?
show the reaction diagram
-
highest transglycosylation efficiency
-
-
?
Galbeta1-6Galbeta1-1'ceramide + H2O
ceramide + Galbeta1-6Galbeta
show the reaction diagram
-
-
-
-
?
Galbeta1-6Galbeta1-6Galbeta1-1'-ceramide + H2O
?
show the reaction diagram
-
-
-
-
?
Galbeta1-6Galbeta1-6Galbeta1-1'-ceramide + H2O
ceramide + Galbeta1-6Galbeta1-6Gal
show the reaction diagram
-
-
-
-
?
Galbeta1-6Galbeta1-6Galbeta1-6Galbeta1-1'-ceramide + H2O
?
show the reaction diagram
-
-
-
-
?
GalNAcalpha(1,4)GalNAcbeta(1,4)GlcNAcbeta(1,3)Manbeta(1,4)Glcbeta-1,1'-ceramide + H2O
ceramide + GalNAcalpha(1,4)GalNAcbeta(1,4)GlcNAcbeta(1,3)Manbeta(1,4)Glc
show the reaction diagram
-
37C, pH 3.0, 0.2% (w/v) Triton X-100
-
-
?
GalNAcalpha(1-3)GalNAcbeta(1-3)Galalpha(1-4)Galbeta(1-4)Glcbeta(1-1)ceramide + H2O
GalNAcalpha(1-3)GalNAcbeta(1-3)Galalpha(1-4)Galbeta(1-4)Glc + ceramide
show the reaction diagram
-
i.e. Gb5Cer
-
-
?
GalNAcbeta(1,3)Galalpha(1,4)Galbeta(1,4)Glcbeta-1,1'-ceramide + H2O
ceramide + GalNAcbeta(1,3)Galalpha(1,4)Galbeta(1,4)Glc
show the reaction diagram
-
37C, pH 3.0, 0.2% (w/v) Triton X-100
-
-
?
GalNAcbeta(1-3)Galalpha(1-4)Galbeta(1-4)Glcbeta(1-1)ceramide + H2O
GalNAcbeta(1-3)Galalpha(1-4)Galbeta(1-4)Glc + ceramide
show the reaction diagram
GalNAcbeta(1-4)(NeuAcalpha(2-3))Galbeta(1-4)Glcbeta(1-1)ceramide + H2O
GalNAcbeta(1-4)(NeuAcalpha(2-3))Galbeta(1-4)Glc + ceramide
show the reaction diagram
GalNAcbeta(1-4)Galbeta(1-4)Glcbeta(1-1)ceramide + H2O
GalNAcbeta(1-4)Galbeta(1-4)Glc + ceramide
show the reaction diagram
-
i.e. Gg3Cer
-
-
?
ganglioside + H2O
?
show the reaction diagram
ganglioside Forssman + H2O
?
show the reaction diagram
-
isoform EGCase I exhibits 25.2% activity with this substrate
-
-
?
ganglioside fucosyl-GM1 + H2O
?
show the reaction diagram
-
100% hydrolyzed by EGCase I and 19% hydrolyzed by EGCase II
-
-
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ganglioside Gb3Cer + H2O
?
show the reaction diagram
-
isoform EGCase I exhibits 100% activity with this substrate
-
-
?
ganglioside Gb4Cer + H2O
?
show the reaction diagram
-
isoform EGCase I exhibits 33.9% activity with this substrate
-
-
?
ganglioside GbOse3Cer + H2O
?
show the reaction diagram
-
73% hydrolyzed by EGCase I and 24% hydrolyzed by EGCase II
-
-
-
ganglioside GbOse4Cer + H2O
?
show the reaction diagram
-
34% hydrolyzed by EGCase I and 7% hydrolyzed by EGCase II
-
-
-
ganglioside GbOse5Cer + H2O
?
show the reaction diagram
-
25% hydrolyzed by EGCase I and 6.5% hydrolyzed by EGCase II
-
-
-
ganglioside GD1a + H2O
?
show the reaction diagram
ganglioside GD1b + H2O
?
show the reaction diagram
ganglioside GD3 + H2O
?
show the reaction diagram
ganglioside GM1 + H2O
?
show the reaction diagram
ganglioside GM1a + H2O
?
show the reaction diagram
-
isoform EGCase I exhibits 100% activity with this substrate
-
-
?
ganglioside GM3 + H2O
?
show the reaction diagram
-
isoform EGCase I exhibits 100% activity with this substrate
-
-
?
ganglioside TGC + H2O
?
show the reaction diagram
-
66% hydrolyzed by EGCase III
-
-
-
GlcA4Me-beta(1,4)[GalNAc-Me-alpha(1,3)]Fucalpha(1,4)GlcNAcbeta(1,2)Manalpha(1,3)[Xylbeta(1,2)]Manbeta(1,4)Glcbeta-1,1'-ceramide + H2O
ceramide + GlcA4Me-beta(1,4)[GalNAc-Me-alpha(1,3)]Fucalpha(1,4)GlcNAcbeta(1,2)Manalpha(1,3)[Xylbeta(1,2)]Manbeta(1,4)Glc
show the reaction diagram
-
37C, pH 3.0, 0.2% (w/v) Triton X-100
-
-
?
Glcbeta1-6Galbeta1-1'-ceramide + H2O
?
show the reaction diagram
-
-
-
-
?
GlcNAcalpha1-2Fucalpha1-3Manbeta1-4Glcbeta1-1'-ceramide + H2O
ceramide + GlcNAcalpha1-2Fucalpha1-3Manbeta1-4Glc
show the reaction diagram
GlcNAcbeta(1-2)Manalpha(1-3)Manbeta(1-4)Glcbeta(1-1)ceramide + H2O
GlcNAcbeta(1-2)Manalpha(1-3)Manbeta(1-4)Glc + ceramide
show the reaction diagram
-
35% of the activity with lactosylceramide
-
?
globotetraosylceramide + H2O
?
show the reaction diagram
-
-
-
-
?
globotriaosylceramide + H2O
?
show the reaction diagram
-
-
-
-
?
glucosylceramide + H2O
?
show the reaction diagram
-
isoform EGCase I exhibits 6.7% activity with this substrate
-
-
?
glycosphingolipid + H2O
?
show the reaction diagram
isoglobotetraosylceramide + H2O
?
show the reaction diagram
-
-
-
-
?
isoglobotriaosylceramide + H2O
?
show the reaction diagram
-
-
-
-
?
lactosylceramide + H2O
?
show the reaction diagram
-
isoform EGCase I exhibits 29.9% activity with this substrate
-
-
?
Manalpha(1-3)Manbeta(1-4)Glcbeta(1-1)ceramide + H2O
Manalpha(1-3)Manbeta(1-4)Glc + ceramide
show the reaction diagram
-
88% of the activity with lactosylceramide
-
?
Manbeta(1-4)Glcbeta(1-1)ceramide + H2O
Manbeta(1-4)Glc + ceramide
show the reaction diagram
-
93% of the activity with lactosylceramide
-
?
monogalactosylceramide + H2O
ceramide + D-galactose
show the reaction diagram
-
-
-
-
?
NeuAcalpha(2,3)Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,3)]Galbeta(1,4)Glcbeta-1,1'-ceramide + H2O
ceramide + NeuAcalpha(2,3)Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,3)]Galbeta(1,4)Glc
show the reaction diagram
-
37C, pH 3.0, 0.2% (w/v) Triton X-100
-
-
?
NeuAcalpha(2,3)Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,8)NeuAcalpha(2,3)]Galbeta(1,4)Glcbeta-1,1'-ceramide + H2O
ceramide + NeuAcalpha(2,3)Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,8)NeuAcalpha-2,3]Gal-beta(1,4)Glc
show the reaction diagram
-
37C, pH 3.0, 0.2% (w/v) Triton X-100, most favoured substrate
-
-
?
NeuAcalpha(2,3)Galbeta(1,4)Glcbeta-1,1'-ceramide + H2O
ceramide + NeuAcalpha(2,3)Galbeta(1,4)Glc
show the reaction diagram
-
37C, pH 3.0, 0.2% (w/v) Triton X-100
-
-
?
NeuAcalpha(2-3)Galbeta(1-3)GalNAcbeta(1-4)(NeuAcalpha(2-3))Galbeta(1-4)Glcbeta(1-1)ceramide + H2O
NeuAcalpha(2-3)Galbeta(1-3)GalNAcbeta(1-4)(NeuAcalpha(2-3))Galbeta(1-4)Glc + ceramide
show the reaction diagram
NeuAcalpha(2-3)Galbeta(1-3)GalNAcbeta(1-4)(NeuAcalpha(2-8)NeuAcalpha(2-3))Galbeta(1-4)Glcbeta(1-1)ceramide + H2O
NeuAcalpha(2-3)Galbeta(1-3)GalNAcbeta(1-4)(NeuAcalpha(2-8)NeuAcalpha(2-3))Galbeta(1-4)Glc + ceramide
show the reaction diagram
NeuAcalpha(2-3)Galbeta(1-4)Glcbeta(1-1)ceramide + H2O
NeuAcalpha(2-3)Galbeta(1-4)Glc + ceramide
show the reaction diagram
NeuAcalpha(2-3)Galbeta(1-4)GlcNAcbeta(1-3)Galbeta(1-4)Glcbeta(1-1)ceramide + H2O
NeuAcalpha(2-3)Galbeta(1-4)GlcNAcbeta(1-3)Galbeta(1-4)Glc + ceramide
show the reaction diagram
NeuAcalpha(2-8)NeuAcalpha(2-3)Galbeta(1-3)GalNAcbeta(1-4)(NeuAcalpha(2-3))Galbeta(1-4)Glcbeta(1-1)ceramide + H2O
NeuAcalpha(2-8)NeuAcalpha(2-3)Galbeta(1-3)GalNAcbeta(1-4)(NeuAcalpha(2-3))Galbeta(1-4)Glc + ceramide
show the reaction diagram
-
GT, 81% of the activity with lactosylceramide
-
?
NeuAcalpha(2-8)NeuAcalpha(2-3)Galbeta(1-3)GalNAcbeta(1-4)(NeuAcalpha(2-8)NeuAcalpha(2-3))Galbeta(1-4)Glcbeta(1-1)ceramide + H2O
NeuAcalpha(2-8)NeuAcalpha(2-3)Galbeta(1-3)GalNAcbeta(1-4)(NeuAcalpha(2-8)NeuAcalpha(2-3))Galbeta(1-4)Glc + ceramide
show the reaction diagram
NeuAcalpha(2-8)NeuAcalpha(2-3)Galbeta(1-4)Glcbeta(1-1)ceramide + H2O
NeuAcalpha(2-8)NeuAcalpha(2-3)Galbeta(1-4)Glc + ceramide
show the reaction diagram
non-acid glycosphingolipids of human gastric epithelium + H2O
?
show the reaction diagram
-
-
-
-
?
oligoglycosylglucosyl-(1<->1)-ceramide + H2O
ceramide + oligoglycosylglucose
show the reaction diagram
-
-
-
-
?
porcine brain ganglioside-1 + H2O
?
show the reaction diagram
-
-
-
-
?
porcine brain ganglioside-3 + H2O
?
show the reaction diagram
-
-
-
-
?
porcine brain ganglioside-4 + H2O
?
show the reaction diagram
-
-
-
-
?
porcine brain ganglioside-5 + H2O
?
show the reaction diagram
-
-
-
-
?
resorufin + beta-D-glucose + H2O
resorufin-beta-D-glucoside
show the reaction diagram
-
highest catalytic efficiency
-
-
?
trigalactosylceramide + H2O
?
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
oligoglycosylglucosyl-(1<->1)-ceramide + H2O
ceramide + oligoglycosylglucose
show the reaction diagram
-
-
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
no significant effect on the activity at 5 mM
Cu2+
-
no significant effect on the activity at 5 mM
Mg2+
-
no significant effect on the activity at 5 mM
Mn2+
-
no significant effect on the activity at 5 mM
Zn2+
-
no significant effect on the activity at 5 mM
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(3R,4R,5R)-4-(beta-D-glucopyranosyl)oxy-3-hydroxy-5-(hydroxymethyl)piperidine
-
-
(5R,6R,7S,8S)-6-(beta-D-glucopyranosyloxy)-5,6,7,8-tetrahydro-5-(hydroxymethyl)imidazo-[1,2-a]pyridine-7,8-diol
-
-
1-(4-dimethylamino)benzoylamino-1,2,5-trideoxy-2,5-imino-d-mannitol
-
-
Ag+
-
1 mM, 63% inhibition of endoglycoceramidase I, 94% inhibition of endoglycoceramidase II
Brij 58
-
-
Ca2+
-
87% inhibition at 5 mM
DMSO
-
inhibits enzyme activity at concentrations higher than 30% (v/v)
ethanol
-
excess amounts inhibit
methanol
-
excess amounts inhibit
SDS
complete inhibition at 0.1%
taurodeoxycholate
Tween 20
-
-
additional information
-
Ca2+, Mg2+, Mn2+, Ni2+, Co2+, and EDTA have no significant effect on the activity
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Activator protein
-
Detergent
-
dimethylformamide
-
the activity is enhanced by addition of 5% (v/v) dimethylformamide
Dimethylsulfoxide
-
the activity is enhanced by addition of 10% (v/v) dimethylsulfoxide
Lubrol Px
Nonidet P-40
sodium cholate
-
-
sodium taurodeoxycholate
-
activates endoglycoceramidase I and II, inhibition of endoglycoceramidase I above 0.4% w/v
taurodeoxycholate
-
0.5 mg/ml, 2.5fold increase of activity
tetrahydrofuran
-
the activity is enhanced by addition of 5% (v/v) tetrahydrofuran
Triton X-100
additional information
-
for maximal hydrolysis of glycosphingolipids by rhodococcal EGCases, a detergent such as Triton X-100 is required at a final concentration of 0.1-0.3% of the reaction mixture. The hydrolysis of water soluble gangliosides such as GM1 does not require the presence of a detergent. For the hydrolysis of GM1, the addition of 5% tetrahydrofuran instead of Triton X-100 increases the reaction rate of EGCase I by 2fold compared to that in the absence of a detergent. Tetrahydrofuran is ineffective in facilitating the hydrolysis of GbOse4Cer by EGCase I. DMSO, ethyl acetate, and methyl cellosolve are also not effective in stimulating the hydrolysis of GM1 or GbOse4Cer by EGCase I in the absence of a detergent
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.4 - 2.9
2-N-hexadecanoylamino-4-nitrophenyl-O-beta-lactoside
0.34
4-methylumbelliferyl beta-D-glucoside
-
isoform EGCrP2, at pH 6.0 and 37C
0.817
4-nitrophenyl-beta-D-glucoside
-
isoform EGCrP2, at pH 6.0 and 37C
0.0058 - 0.0384
C6-7-nitro-2,1,3-benzoxadiazole-glucosylceramide
-
0.3
D-erythro-dihydrosphingosine
-
-
0.04
D-erythro-sphingosine
-
-
0.15 - 0.35
Galbeta(1-3)GalNAcbeta(1-4)(NeuAcalpha(2-3))Galbeta(1-4)Glcbeta(1-1)ceramide
0.28 - 16.5
Galbeta(1-3)GalNAcbeta(1-4)(NeuAcalpha(2-8)NeuAcalpha(2-3))Galbeta(1-4)Glcbeta(1-1)ceramide
0.1 - 0.5
Galbeta(1-3)GalNAcbeta(1-4)Galbeta(1-4)Glcbeta(1-1)ceramide
0.43
Galbeta1-6Galbeta1-6Galbeta1-1'ceramide
-
-
0.047
GalNAcbeta(1-4)Galbeta(1-4)Glcbeta(1-1)ceramide
-
i.e. Gg3Cer from guinea pig erythrocyte cell surface, pH 7, 37C
0.23 - 0.46
ganglioside Gb3Cer
0.4 - 0.43
ganglioside GM1a
0.044
NeuAcalpha(2-3)Galbeta(1-4)Glcbeta(1-1)ceramide
-
from horse erythrocyte cell surface, pH 7, 37C
0.0436
resorufin
-
isoform EGCrP2, at pH 6.0 and 37C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
8.8
4-methylumbelliferyl beta-D-glucoside
Cryptococcus neoformans
-
isoform EGCrP2, at pH 6.0 and 37C
13.8
4-nitrophenyl-beta-D-glucoside
Cryptococcus neoformans
-
isoform EGCrP2, at pH 6.0 and 37C
0.0383 - 0.53
C6-7-nitro-2,1,3-benzoxadiazole-glucosylceramide
-
0.12
D-erythro-dihydrosphingosine
Rhodococcus sp.
-
-
0.07
D-erythro-sphingosine
Rhodococcus sp.
-
-
13.3
Galbeta1-6Galbeta1-6Galbeta1-1'ceramide
Rhodococcus equi
-
-
0.069 - 85
ganglioside Gb3Cer
5.7 - 120
ganglioside GM1a
21.9
resorufin
Cryptococcus neoformans
-
isoform EGCrP2, at pH 6.0 and 37C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
25.7
4-methylumbelliferyl beta-D-glucoside
Cryptococcus neoformans
-
isoform EGCrP2, at pH 6.0 and 37C
1709
16.9
4-nitrophenyl-beta-D-glucoside
Cryptococcus neoformans
-
isoform EGCrP2, at pH 6.0 and 37C
3388
6.6 - 13.8
C6-7-nitro-2,1,3-benzoxadiazole-glucosylceramide
202165
0.3 - 190
ganglioside Gb3Cer
202166
13 - 320
ganglioside GM1a
16087
504
resorufin
Cryptococcus neoformans
-
isoform EGCrP2, at pH 6.0 and 37C
37383
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.005
(3R,4R,5R)-4-(beta-D-glucopyranosyl)oxy-3-hydroxy-5-(hydroxymethyl)piperidine
-
-
0.0005
(5R,6R,7S,8S)-6-(beta-D-glucopyranosyloxy)-5,6,7,8-tetrahydro-5-(hydroxymethyl)imidazo-[1,2-a]pyridine-7,8-diol
-
-
0.01
1-(4-dimethylamino)benzoylamino-1,2,5-trideoxy-2,5-imino-d-mannitol
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0000032
Marphysa sp.
-
37C, pH 3.0, 0.2% (w/v) Triton X-100, with Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,3)]Galbeta(1,4)Glcbeta-1,1'-ceramide as substrate
0.0000044
Meretrix sp.
-
37C, pH 3.0, 0.2% (w/v) Triton X-100, with Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,3)]Galbeta(1,4)Glcbeta-1,1'-ceramide as substrate
0.0000046
Ruditapes sp.
-
37C, pH 3.0, 0.2% (w/v) Triton X-100, with Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,3)]Galbeta(1,4)Glcbeta-1,1'-ceramide as substrate
0.0000058
-
37C, pH 3.0, 0.2% (w/v) Triton X-100, with Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,3)]Galbeta(1,4)Glcbeta-1,1'-ceramide as substrate
0.0000118
Cellana sp.
-
37C, pH 3.0, 0.2% (w/v) Triton X-100, with Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,3)]Galbeta(1,4)Glcbeta-1,1'-ceramide as substrate
0.0000747
37C, pH 3.0, 0.2% (w/v) Triton X-100, with Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,3)]Galbeta(1,4)Glcbeta-1,1'-ceramide as substrate
0.00021
-
gastric cavity, 37C, pH 3.0, 0.2% (w/v) Triton X-100, with Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,3)]Galbeta(1,4)Glcbeta-1,1'-ceramide as substrate
0.00023
-
peduncle regions, 37C, pH 3.0, 0.2% (w/v) Triton X-100, with Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,3)]Galbeta(1,4)Glcbeta-1,1'-ceramide as substrate
0.0002325
-
37C, pH 3.0, 0.2% (w/v) Triton X-100, with Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,3)]Galbeta(1,4)Glcbeta-1,1'-ceramide as substrate
0.0002346
Entacmaea sp.
-
37C, pH 3.0, 0.2% (w/v) Triton X-100, with Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,3)]Galbeta(1,4)Glcbeta-1,1'-ceramide as substrate
0.0002403
Aiptasia sp.
-
37C, pH 3.0, 0.2% (w/v) Triton X-100, with Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,3)]Galbeta(1,4)Glcbeta-1,1'-ceramide as substrate
0.00025
-
tentacles, 37C, pH 3.0, 0.2% (w/v) Triton X-100, with Galbeta(1,3)GalNAcbeta(1,4)[NeuAcalpha(2,3)]Galbeta(1,4)Glcbeta-1,1'-ceramide as substrate
23.5
-
isoform I
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 6.5
-
-
5.5 - 6
-
isoform EGCase I
6.5
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 8
-
about 85% activity at pH 4.0, 100% activity at pH 5.0, about 70% activity at pH 6.0, about 55% activity at pH 7.0, about 30% activity at pH 8.0
7
-
90% activity for NeuAcalpha(2-3)Galbeta(1-4)Glcbeta(1-1)ceramide, 64% activity for Gg3Cer
additional information
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15 - 40
-
about 45% activity at 15C, about 65% activity at 25C, about 75% activity at 30C, 100% activity at 37C, about 78% activity at 40C
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.5
-
endoglycoceramidase I
4
-
pH gradient from pH 3 to pH 10
4.28
-
sequence analysis
4.5
-
endoglycoceramidase II
5.47
-
calculated from amino acid sequence
5.61
calculated from amino acid sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
-
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
32700
-
endoglycoceramidase I, gel filtration
43000
-
endoglycoceramidase II, gel filtration
51000
gel filtration
52299
-
x * 54000, SDS-PAGE, x * 52299, sequence analysis
52900
-
after treatment with glycopeptidase F, monomer, SDS-PAGE
54000
-
x * 54000, SDS-PAGE, x * 52299, sequence analysis
55000
-
x * 55000, isoform EGCase I, SDS-PAGE
55900
-
endoglycoceramidase I, PAGE in absence and presence of SDS
56000
-
monomer, SDS-PAGE
58900
-
endoglycoceramidase II, PAGE in absence and presence of SDS
59680
-
calculated from amino acid sequence
65000
-
x * 65000, SDS-PAGE
250000 - 450000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
enzyme contains 6 potential N-glycosylation sites
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by vapor diffusion method, crystals of the native enzyme belong to space group P21 with unit cell dimensions a=53.8 A, b=92.9 , c=94.5 A,beta=98.6, to 1.6 A resolution. Crystal structure of mutant E351S in complex with the ganglioside GM3, crystals belong to space group C2, with unit cell dimensions a = 77.8 A, b = 62.0 A, c = 102.8 A, beta = 112.3, to 1.1 A resolution. Active site of endo-glycoceramidase is split into a wide, polar cavity to bind the polyhydroxylated oligosaccharide moiety and a narrow, hydrophobic tunnel to bind the ceramide lipid chains
-
EGC-inhibitor complexes, to 1.50-1.85 A resolution. Both (3R,4R,5R)-4-(beta-D-glucopyranosyl)oxy-3-hydroxy-5-(hydroxymethyl)piperidine and (5R,6R,7S,8S)-6-(beta-D-glucopyranosyloxy)-5,6,7,8-tetrahydro-5-(hydroxymethyl)imidazo-[1,2-a]pyridine-7,8-diol bind in the -2 and -1 subsites of the enzyme, with the isofagomine and glucoimidazole moieties located in the catalytic -1 subsite. 1-(4-dimethylamino)benzoylamino-1,2,5-trideoxy-2,5-imino-d-mannitol binds in the -1 subsite in an envelope conformation. The ring nitrogen superimposes with the anomeric carbon of the lactosyl-enzyme intermediate and the ring nitrogen of the isofagomine
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.5 - 8
-
stable
646688
4 - 8
-
stable for 60 min
646691
4.5 - 8
-
37C, 30 min, stable
646688
5 - 9
-
stable
646687
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
60 min, pH 3, 50.4% of endoglycoceramidase I activity and 27.5% of endoglycoceramidase II activity lost; 60 min pH 4-8, endoglycoceramidase I and II, stable
45
-
pH 7.0, 10 min, stable up to
60
-
pH 7.0, 10 min, complete inactivation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
highly purified ECGase II loses activity due to instability following purification
-
stable to repeated freezing and thawing
-
unstable at protein concentration below 0.2 mg/ml or without Triton X-100
-
unstable to repeated freezing and thawing
-
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
the enzyme remains stable in up to 30% (v/v) DMSO (more than 80% activity)
-
732178
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, stable for at least 1 year
-
0C, protein concentration greater than 0.2 mg/ml, 50 mM sodium acetate buffer at pH 6.0 or 20 mM phosphate buffer at pH 7.2, 0.1-1.0% Triton X-100, stable for several months
-
frozen, stable for at least 6 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2 molecular species: EGCaseI and EGCaseII, homogeneity
-
by gel filtration, 2100fold
-
by Ni(II) affinity chromatography
-
by Ni(II) affinity chromatography to more than 95% purity
-
EGC and its mutants
-
EGCase and EGCase P
EGCase II
-
homogeneity
-
Ni Sepharose 6 column chromatography
-
Ni Sepharose 6 column chromatography and Superdex 200 gel filtration
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
amplified gene product encoding amino acid residues 23-488 without the signal peptide digested with HindIII and NotI and inserted into the corresponding sites of pET22b(+) to generate a C-terminal His6-tagged protein. The recombinant plasmid and mutants expressed in Escherichia coli strain BL21(DE3)
-
EGCase and EGCase P
expressed in Escherichia coli BL21(DE3) cells
expressed in Rhodococcus erythropolis JCM3201 cells
-
expression in Chinese hamster ovary cells and Escherichia coli
-
mutant E351S in pET28a subcloned into pTKNd and pUC18 at the NdeI/SalI and EcoRI/SalI restriction sites, respectively. EGC and its mutants expressed in Escherichia coli BL21 (Tuner) cells
-
recombinant EGC, lacking the 30 amino acid N-terminal signal sequence overexpressed in Escherichia coli
-
recombinant protein from strain C9
-
subcloned into pET28a using the NdeI/XhoI restriction sites and expressed in Escherichia coli BL21
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E234D
-
strongly decreased enzyme activity compared with the wild-type enzyme
E234Q
-
no activity
E341D
-
strongly decreased enzyme activity compared with the wild-type enzyme
E341Q
-
no activity
E224D
-
strain C9, severely reduced enzyme activity
E224Q
-
strain C9, severely reduced enzyme activity
E233A
-
glycosyl-enzyme intermediate trapped by soaking crystals of the general acid/base knock-out mutant E233A, with an activated sugar donor, 2,4-dinitrophenyl beta-D-lactoside
E233D
-
strain M-777, severely reduced enzyme activity
E233Q
-
strain M-777, severely reduced enzyme activity
E351A
-
mutant glycosidase with glycosynthase activity
E351G
-
mutant glycosidase with glycosynthase activity
E351M
-
mutant glycosidase with no glycosynthase activity
E351R
-
mutant glycosidase with no glycosynthase activity
E351W
-
mutant glycosidase with no glycosynthase activity
E351Y
-
mutant glycosidase with no glycosynthase activity
E224D
-
strain C9, severely reduced enzyme activity
-
E224Q
-
strain C9, severely reduced enzyme activity
-
E233D
-
strain M-777, severely reduced enzyme activity
-
E233Q
-
strain M-777, severely reduced enzyme activity
-
E224D
-
strain C9, severely reduced enzyme activity
-
E224Q
-
strain C9, severely reduced enzyme activity
-
E233A
-
glycosyl-enzyme intermediate trapped by soaking crystals of the general acid/base knock-out mutant E233A, with an activated sugar donor, 2,4-dinitrophenyl beta-D-lactoside
-
E233D
-
strain M-777, severely reduced enzyme activity
-
E233Q
-
strain M-777, severely reduced enzyme activity
-
E351A
-
mutant glycosidase with glycosynthase activity
-
E351G
-
mutant glycosidase with glycosynthase activity
-
E351M
-
mutant glycosidase with no glycosynthase activity
-
E351R
-
mutant glycosidase with no glycosynthase activity
-
E351S
-
mutant glycosidase with glycosynthase activity, most active enzyme for ganglioside synthesis; prevents substrate hydrolysis during crystallization; the endo-glycoceramidase II nucleophile mutant E351S has glycosynthase activity
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
drug development
medicine
-
release of glycans from the ceramide moieties of glycosphingolipids by endoglycoceramidase II treatment in order to analyze structures of glycosphingolipids in normal human colorectal epithelial cells, and characteristic alterations of oligosaccharide structures in malignant transformation