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Literature summary for 3.2.1.97 extracted from
- Crystallographic and mutational analyses of substrate recognition of endo-alpha-N-acetylgalactosaminidase from Bifidobacterium longum (2009), J. Biochem., 146, 389-398.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression of His-tagged truncated EngBF mutants, comprising residues 340-1528 and 340-1694, respectively, and point mutants in Escherichia coli strain BL21(DE3) | Bifidobacterium longum |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| purified recombinant His-tagged enzyme, X-ray diffraction structure analysis at 2.0-2.5 A resolution | Bifidobacterium longum |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D1295A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the truncated wild-type enzyme | Bifidobacterium longum |
| D682A | site-directed mutagenesis, the mutant shows reduced activity compared to the truncated wild-type enzyme | Bifidobacterium longum |
| D789A | site-directed mutagenesis, the mutant shows reduced activity compared to the truncated wild-type enzyme | Bifidobacterium longum |
| E822A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the truncated wild-type enzyme | Bifidobacterium longum |
| K1199A | site-directed mutagenesis, the mutant shows increased activity compared to the truncated wild-type enzyme | Bifidobacterium longum |
| N720A | site-directed mutagenesis, the mutant shows increased activity compared to the truncated wild-type enzyme | Bifidobacterium longum |
| Q894A | site-directed mutagenesis, the mutant shows increased activity compared to the truncated wild-type enzyme | Bifidobacterium longum |
| W748A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the truncated wild-type enzyme | Bifidobacterium longum |
| W748F | site-directed mutagenesis, the mutant shows highly reduced activity compared to the truncated wild-type enzyme | Bifidobacterium longum |
| W748Y | site-directed mutagenesis, the mutant shows highly reduced activity compared to the truncated wild-type enzyme | Bifidobacterium longum |
| W750A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the truncated wild-type enzyme | Bifidobacterium longum |
| W750F | site-directed mutagenesis, the mutant shows reduced activity compared to the truncated wild-type enzyme | Bifidobacterium longum |
| W750Y | site-directed mutagenesis, the mutant shows reduced activity compared to the truncated wild-type enzyme | Bifidobacterium longum |
| Y787F | site-directed mutagenesis, the mutant shows highly reduced activity compared to the truncated wild-type enzyme | Bifidobacterium longum |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Ca2+ | - |
Bifidobacterium longum |  |
| Cd2+ | strong inhibition at 5 mM | Bifidobacterium longum | |
| EDTA | - |
Bifidobacterium longum | |
| Li+ | - |
Bifidobacterium longum | |
| Zn2+ | strong inhibition at 5 mM | Bifidobacterium longum | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Co2+ | activates at 5 mM | Bifidobacterium longum | |
| Mg2+ | activates at 5 mM | Bifidobacterium longum | |
| Mn2+ | activates at 5 mM | Bifidobacterium longum | |
| Ni2+ | activates at 5 mM | Bifidobacterium longum | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | Bifidobacterium longum | the enzyme hydrolyses the O-glycosidic bonds in mucin-type O-glycan between alpha-GalNAc and Ser/Thr. EngBF is highly specific for the core 1-type O-glycan to release the disaccharide Galbeta1-3GalNAc | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Bifidobacterium longum | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged EngBF truncation and point mutants from Escherichia coli strain BL21(DE3) by nickel affinity and anion exchange chromatography, and gel filtration | Bifidobacterium longum |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | the enzyme hydrolyses the O-glycosidic bonds in mucin-type O-glycan between alpha-GalNAc and Ser/Thr. EngBF is highly specific for the core 1-type O-glycan to release the disaccharide Galbeta1-3GalNAc | Bifidobacterium longum | ? | - |
? | |
| additional information | active site structure and substrate binding by the enzyme, important residues for substrate binding are Trp residues Trp748 and Trp750, appearing to form stacking interactions with the beta-faces of sugar rings of Galbeta1-3GalNAc by substrate-induced fit, substrate specificity for glycans, docking analysis, detailed overview | Bifidobacterium longum | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| More | docking analysis and structural model building, overview | Bifidobacterium longum |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| endo-alpha-GalNAc-ase | - |
Bifidobacterium longum |
| endo-alpha-N-acetylgalactosaminidase | - |
Bifidobacterium longum |
| EngBF | - |
Bifidobacterium longum |
| More | the enzyme belongs to the glycoside hydrolase family 101, GH101 | Bifidobacterium longum |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 60 | - |
assay at | Bifidobacterium longum |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
assay at | Bifidobacterium longum |
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