EC Number | Application | Comment | Organism |
---|---|---|---|
3.2.1.123 | drug development | rational redesign of EGC toward the synthesis of novel ganglioside-derived therapeutics | Rhodococcus sp. |
EC Number | Cloned (Comment) | Organism |
---|---|---|
3.2.1.62 | expression of the gene lacking the 30-residue N-terminal secretion signal sequence in Escherichia coli, selenomethionine-derivatized enzyme | Rhodococcus sp. |
3.2.1.123 | subcloned into pET28a using the NdeI/XhoI restriction sites and expressed in Escherichia coli BL21 | Rhodococcus sp. |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
3.2.1.62 | 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 | Rhodococcus sp. |
3.2.1.123 | 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 Å, 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 | Rhodococcus sp. |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
3.2.1.62 | E233A | site directed mutagenesis | Rhodococcus sp. |
3.2.1.62 | E351S | site directed mutagenesis | Rhodococcus sp. |
3.2.1.123 | 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 | Rhodococcus sp. |
3.2.1.123 | E351S | prevents substrate hydrolysis during crystallization | Rhodococcus sp. |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.2.1.62 | 0.04 | - |
D-erythro-sphingosine | - |
Rhodococcus sp. | |
3.2.1.62 | 0.3 | - |
D-erythro-dihydrosphingosine | - |
Rhodococcus sp. | |
3.2.1.123 | 0.04 | - |
D-erythro-sphingosine | - |
Rhodococcus sp. | |
3.2.1.123 | 0.3 | - |
D-erythro-dihydrosphingosine | - |
Rhodococcus sp. |
EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|---|
3.2.1.123 | membrane | - |
Rhodococcus sp. | 16020 | - |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.2.1.62 | Rhodococcus sp. | - |
- |
- |
3.2.1.123 | Rhodococcus sp. | - |
- |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
3.2.1.62 | of the recombinant protein by by Ni(II) affinity chromatography | Rhodococcus sp. |
3.2.1.123 | by Ni(II) affinity chromatography to more than 95% purity | Rhodococcus sp. |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.2.1.62 | D-erythro-dihydrosphingosine + a sugar | - |
Rhodococcus sp. | ? | - |
? | |
3.2.1.62 | D-erythro-sphingosine + a sugar | - |
Rhodococcus sp. | ? | - |
? | |
3.2.1.123 | D-erythro-dihydrosphingosine + H2O | - |
Rhodococcus sp. | ? | - |
? | |
3.2.1.123 | D-erythro-sphingosine + H2O | - |
Rhodococcus sp. | ? | - |
? | |
3.2.1.123 | ganglioside + H2O | - |
Rhodococcus sp. | ? | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
3.2.1.62 | EGC | - |
Rhodococcus sp. |
3.2.1.62 | endo-glycoceramidase II | membrane-associated family 5 glycosidase | Rhodococcus sp. |
3.2.1.62 | glycosylceramidase | - |
Rhodococcus sp. |
3.2.1.123 | EGC | - |
Rhodococcus sp. |
3.2.1.123 | endo-glycoceramidase II | - |
Rhodococcus sp. |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.2.1.123 | 0.07 | - |
D-erythro-sphingosine | - |
Rhodococcus sp. | |
3.2.1.123 | 0.12 | - |
D-erythro-dihydrosphingosine | - |
Rhodococcus sp. |