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Literature summary for 3.2.1.62 extracted from

  • Ishibashi, Y.; Ikeda, K.; Sakaguchi, K.; Okino, N.; Taguchi, R.; Ito, M.
    Quality control of fungus-specific glucosylceramide in Cryptococcus neoformans by endoglycoceramidase-related protein 1 (EGCrP1) (2012), J. Biol. Chem., 287, 368-381.
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
EDTA about 120% activity at 1 mM Cryptococcus neoformans

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Cryptococcus neoformans

Protein Variants

Protein Variants Comment Organism
E254Q inactive Cryptococcus neoformans
E483Q inactive Cryptococcus neoformans

Inhibitors

Inhibitors Comment Organism Structure
Ca2+ about 78% residual activity at 1 mM Cryptococcus neoformans
Co2+ about 18% residual activity at 1 mM Cryptococcus neoformans
Cu2+ about 10% residual activity at 1 mM Cryptococcus neoformans
DMSO the enzyme is inhibited by 30-50% (v/v) DMSO Cryptococcus neoformans
Fe2+ about 40% residual activity at 1 mM Cryptococcus neoformans
Fe3+ about 50% residual activity at 1 mM Cryptococcus neoformans
Hg+ complete inhibition at 1 mM Cryptococcus neoformans
Mg2+ about 90% residual activity at 1 mM Cryptococcus neoformans
Mn2+ about 50% residual activity at 1 mM Cryptococcus neoformans
Na+ about 93% residual activity at 1 mM Cryptococcus neoformans
Ni2+ about 5% residual activity at 1 mM Cryptococcus neoformans
Pb2+ about 10% residual activity at 1 mM Cryptococcus neoformans
Triton X-100 strongly inhibits the activity even at a low concentration Cryptococcus neoformans
Zn2+ about 8% residual activity at 1 mM Cryptococcus neoformans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0058
-
C6-7-nitro-2,1,3-benzoxadiazole-glucosylceramide at pH 7.3 and 30°C Cryptococcus neoformans

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane
-
Cryptococcus neoformans 16020
-

Metals/Ions

Metals/Ions Comment Organism Structure
additional information no metal ions are required for the activity of isoform EGCrP1 Cryptococcus neoformans

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
oligoglycosylglucosyl-(1->1)-ceramide + H2O Cryptococcus neoformans
-
ceramide + oligoglycosylglucose
-
?
oligoglycosylglucosyl-(1->1)-ceramide + H2O Cryptococcus neoformans H99
-
ceramide + oligoglycosylglucose
-
?

Organic Solvent Stability

Organic Solvent Comment Organism
DMSO the enzyme is stimulated by 10-20% (v/v) DMSO Cryptococcus neoformans

Organism

Organism UniProt Comment Textmining
Cryptococcus neoformans H1AE12
-
-
Cryptococcus neoformans H99 H1AE12
-
-

Purification (Commentary)

Purification (Comment) Organism
GSTrap column chromatography Cryptococcus neoformans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
C6-7-nitro-2,1,3-benzoxadiazole-glucosylceramide + H2O highest activity Cryptococcus neoformans ?
-
?
C6-7-nitro-2,1,3-benzoxadiazole-glucosylceramide + H2O highest activity Cryptococcus neoformans H99 ?
-
?
glucosylceramide + H2O low activity Cryptococcus neoformans ?
-
?
glucosylceramide + H2O low activity Cryptococcus neoformans H99 ?
-
?
oligoglycosylglucosyl-(1->1)-ceramide + H2O
-
Cryptococcus neoformans ceramide + oligoglycosylglucose
-
?
oligoglycosylglucosyl-(1->1)-ceramide + H2O
-
Cryptococcus neoformans H99 ceramide + oligoglycosylglucose
-
?

Synonyms

Synonyms Comment Organism
EGCrP1
-
Cryptococcus neoformans
endoglycoceramidase-related protein 1
-
Cryptococcus neoformans
glucocerebrosidase
-
Cryptococcus neoformans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30 35
-
Cryptococcus neoformans

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
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 Cryptococcus neoformans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0383
-
C6-7-nitro-2,1,3-benzoxadiazole-glucosylceramide at pH 7.3 and 30°C Cryptococcus neoformans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
Cryptococcus neoformans

General Information

General Information Comment Organism
malfunction gene disruption results in the accumulation of fungus-specific glucosylceramide and immature glucosylceramide that possess sphingoid bases without a methyl substitution concomitant with a dysfunction of polysaccharide capsule formation Cryptococcus neoformans
metabolism the enzyme participates in the catabolism of glucosylceramide and especially functions to eliminate immature glucosylceramide in vivo that are generated as by-products due to the broad specificity of glucosylceramide synthase Cryptococcus neoformans

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
6.6
-
C6-7-nitro-2,1,3-benzoxadiazole-glucosylceramide at pH 7.3 and 30°C Cryptococcus neoformans