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Literature summary extracted from

  • Ben Bdira, F.; Jiang, J.; Kallemeijn, W.; de Haan, A.; Florea, B.I.; Bleijlevens, B.; Boot, R.; Overkleeft, H.S.; Aerts, J.M.; Ubbink, M.
    Hydrophobic interactions contribute to conformational stabilization of endoglycoceramidase II by mechanism-based probes (2016), Biochemistry, 55, 4823-4835 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.62 expressed in Escherichia coli BL21(DE3) pLysS cells Rhodococcus sp. M777

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.62 hanging drop vapor diffusion method, using 1% (v/v) Triton X-100, 10% (v/v) glycerol, and 25% (w/v) PEG Rhodococcus sp. M777

General Stability

EC Number General Stability Organism
3.2.1.123 the stability of EGCII is markedly enhanced by formation of covalent complexes with cyclophellitol activity-based probes substituted with hydrophobic moieties, as evidenced by an increased melting temperature, resistance against tryptic digestion, changes in 15N-1H transverse relaxation optimized spectroscopy spectra of the [15N]Leu-labeled enzyme, and relative hydrophobicity as determined by 8-anilino-1-naphthalenesulfonic acid fluorescence. The stabilization of EGCII conformation correlates with the shape and hydrophobicity of the substituents of the activity-based probes Rhodococcus sp.

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.62 nonyl-deoxynojirimycin
-
 Rhodococcus sp. M777
3.2.1.123 additional information beta-glucoside-configured cyclophellitol-type activity-based probes are effective mechanism-based, and irreversible inhibitors Rhodococcus sp.

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.62 GM3 ganglioside + H2O Rhodococcus sp. M777
-
 ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.62 Rhodococcus sp. M777 O33853
-
-
3.2.1.123 Rhodococcus sp. O33853
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.62 nickel affinity column chromatography Rhodococcus sp. M777

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.62 4-methylumbelliferyl beta-D-lactopyranoside + H2O
-
 Rhodococcus sp. M777 4-methylumbelliferol + beta-D-lactopyranose
-
 ?
3.2.1.62 GM3 ganglioside + H2O
-
 Rhodococcus sp. M777 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.62 monomer or dimer x * 34000 + x * 25000, SDS-PAGE Rhodococcus sp. M777

Synonyms

EC Number Synonyms Comment Organism
3.2.1.62 EGCII
-
 Rhodococcus sp. M777
3.2.1.62 endoglycoceramidase II
-
 Rhodococcus sp. M777
3.2.1.123 EGCII
-
 Rhodococcus sp.
3.2.1.123 endoglycoceramidase II
-
 Rhodococcus sp.

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.62 37
-
-
 Rhodococcus sp. M777

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.62 25
-
 after 1 h at 25°C in 150mM McIlvaine's buffer (pH 5.5), the enzyme remains stable with no changes in its secondary structure and its active state Rhodococcus sp. M777
3.2.1.62 42
-
 melting temperature Rhodococcus sp. M777

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.62 5.5
-
 in 150 mM McIlvaine's buffer Rhodococcus sp. M777