Information on EC 3.5.1.69 - glycosphingolipid deacylase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.5.1.69
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RECOMMENDED NAME
GeneOntology No.
glycosphingolipid deacylase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
hydrolysis of gangliosides and neutral glycosphingolipids, releasing fatty acids to form the lyso-derivatives
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of linear amides
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SYSTEMATIC NAME
IUBMB Comments
glycosphingolipid amidohydrolase
Does not act on sphingolipids such as ceramide. Not identical with EC 3.5.1.23 ceramidase.
CAS REGISTRY NUMBER
COMMENTARY hide
122544-53-0
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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-
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Manually annotated by BRENDA team
commercial preparation
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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
16-(acetylsulfanyl)hexadecanoic acid + lyso-GM1a
? + H2O
show the reaction diagram
16-[(prop-2-yn-1-yl)oxy]hexadecanoic acid + lyso-GM1a
? + H2O
show the reaction diagram
2-hydroxydodecanoic acid + lyso-GM1a
? + H2O
show the reaction diagram
2-hydroxyhexadecanoic acid + lyso-GM1a
? + H2O
show the reaction diagram
arachidic acid + lyso-GM1a
? + H2O
show the reaction diagram
asialoganglioside GM1 + H2O
fatty acid + ?
show the reaction diagram
behenic acid + lyso-GM1a
? + H2O
show the reaction diagram
caproic acid + lyso-GM1a
? + H2O
show the reaction diagram
caprylic acid + lyso-GM1a
? + H2O
show the reaction diagram
D-galactosylceramide-3-O-sulfate + H2O
?
show the reaction diagram
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-
-
-
?
decanoic acid + lyso-GM1a
? + H2O
show the reaction diagram
docosahexaenoic acid + lyso-GM1a
? + H2O
show the reaction diagram
eicosapentaenoic acid + lyso-GM1a
? + H2O
show the reaction diagram
galacosylceramide + H2O
?
show the reaction diagram
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-
-
-
?
ganglioside asialo-GM1 + H2O
?
show the reaction diagram
ganglioside asialo-GM2 + H2O
?
show the reaction diagram
-
-
-
-
?
ganglioside Gb3-ceramide + H2O
?
show the reaction diagram
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-
-
-
?
ganglioside GD1a + H2O
fatty acid + lyso-GD1a-ganglioside
show the reaction diagram
ganglioside GD1b + H2O
fatty acid + ?
show the reaction diagram
ganglioside GM1 + H2O
fatty acid + lyso-GM1-ganglioside
show the reaction diagram
ganglioside GM1a + H2O
fatty acid + ?
show the reaction diagram
ganglioside GM2 + H2O
fatty acid + lyso-GM2-ganglioside
show the reaction diagram
ganglioside GM3 + H2O
fatty acid + ?
show the reaction diagram
ganglioside GM3 + H2O
fatty acid + lyso-GM3-ganglioside
show the reaction diagram
ganglioside GQ1b + H2O
?
show the reaction diagram
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-
-
-
?
ganglioside GT1b + H2O
fatty acid + ?
show the reaction diagram
gangliotetraosylceramide + H2O
glucosylceramide
show the reaction diagram
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immobilised enzyme, pH 5, 37C. The rate of hydrolysis of the substrate by the immobilised enzyme is 62% after 20 h as compared to 48% with the soluble enzyme under the same reaction conditions
using the soluble form of the SCDase C17:0 glucosylceramide is prepared in bulk solution. Mass spectra reveal a considerable amount of contaminants in the final product. The contaminants are identified as glucosylceramide isoforms with C16:0 and C18:0 fatty acids. The total yield of glucosylceramide is 99%. Products are analyzed by HPTLC
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?
globotetraosylceramide + H2O
?
show the reaction diagram
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?
glucosylceramide + H2O
?
show the reaction diagram
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?
glycosphingolipid + H2O
?
show the reaction diagram
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-
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?
GM1a + H2O
stearic acid + lyso-GM1a
show the reaction diagram
GM3 + H2O
stearic acid + lyso-GM3
show the reaction diagram
hexadecanedioic acid + lyso-GM1a
? + H2O
show the reaction diagram
hexadecanedioic acid methyl ester + lyso-GM1a
? + H2O
show the reaction diagram
lactosylceramide + H2O
?
show the reaction diagram
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-
-
?
lauric acid + lyso-GM1a
? + H2O
show the reaction diagram
lignoceric acid + lyso-GM1a
? + H2O
show the reaction diagram
myristic acid + lyso-GM1a
? + H2O
show the reaction diagram
oleic acid + lyso-GM1a
? + H2O
show the reaction diagram
omega-azido palmitic acid + lyso-GM1a
? + H2O
show the reaction diagram
omega-bromo-palmitic acid + lyso-GM1a
? + H2O
show the reaction diagram
omega-hydroxypalmitic acid + lyso-GM1a
? + H2O
show the reaction diagram
palmitic acid + lyso-GM1a
? + H2O
show the reaction diagram
sphingomyelin + H2O
?
show the reaction diagram
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?
additional information
?
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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
glycosphingolipid + H2O
?
show the reaction diagram
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-
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?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ba2+
-
10 mM, slight activation
Co2+
promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme
Fe2+
promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme
Ni2+
promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Ca2+
promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme
Co2+
promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme
deoxycholic acid
complete inhibition of the hydrolytic and synthetic activity
dimethoxyethane
addition of 5% of DME strongly promoted the activity, further increasing the DME concentrations progressively inhibited synthetic activity
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DMSO
low concentrations of DMSO, up to a maximum at 10%, enhance synthetic activity, while higher concentrations inhibit
Fe2+
promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme
Mg2+
promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme
Ni2+
promotes the hydrolytic activity but inhibits the synthetic activity of the enzyme
Sn2+
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10 mM, strong
taurodeoxycholate
complete inhibition of the hydrolytic and synthetic activity
Triton X-100
activates the hydrolytic reaction, best at 0.5% w/v, but inhibits the synthetic reaction
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
sodium taurodeoxycholate
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1.0 mg/ml, slight activation
taurodeoxycholate
may substitute for Triton X-100 at 0.5%
Triton X-100
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30.4
pH 6.0, 37C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 9
activity range, hydrolysis
5.5 - 10
activity range, synthesis
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
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tightly bound to
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
52000
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x * 52000, SDS-PAGE
75000
1 * 109800, deduced from gene sequence, 1 * 75000, SDS-PAGE
109800
1 * 109800, deduced from gene sequence, 1 * 75000, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 52000, SDS-PAGE
monomer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 9
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stable
288938
7.2
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45C, 30 min, stable
171986
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45
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pH 7.2, 30 min, stable
55
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pH 7.2, 30 min, 50% loss of activity
60
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30 min, 20% loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
SCDase immobilised on magnetic macroporous cellulose maintains its activity without loss after 15 uses
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-85C, 2 months, stable
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4C, 50 mM phosphate buffer, pH 7, 0.1% Triton X-100, long-term stability is achieved by immobilisation on magnetic macroporous cellulose. The enzyme retains the same activity after 1.5 years in storage buffer
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged enzyme, lacking the secretion signal, from Escherichia coli strain BL21 (DE3) pLysS by nickel affinity chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
the gene encoding the mature SA_SCD protein, which lacks its 38-residue N-terminal secretion signal sequence and from which the 277-residue C-terminal sequence is deleted, is codon-optimized and expressed in Escherichia coli strain BL21 (DE3) pLysS as Histagged enzyme
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
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the enzyme is immobilised on magnetic macroporous cellulose and is used to semisynthesise C17:0 glucosylceramide and C17:0 sulfatide, which are required as internal standards for quantification of the corresponding glycosphingolipids by tandem mass spectrometry
synthesis