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Literature summary for 3.2.1.21 extracted from
- The crystal structure of human cytosolic beta-glucosidase unravels the substrate aglycone specificity of a family 1 glycoside hydrolase (2007), J. Mol. Biol., 370, 964-975.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression of wild-type and mutant enzymes in Pichia pastoris | Homo sapiens |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| purified wild-type enzyme, 0.002 ml of protein solution containing 4 mg/ml protein in 20 mM MES, pH 6.0, are mixed with 0.001 ml of reservoir solution containing 0.2 M ammonium sulphate, MES buffer, pH 6.5, and 30% w/v polyethyleneglycol-monomethylether 5500, 20°C, several days, X-ray diffraction structure determination and anaylsis at 2.7 A resolution, molecular replacement method, modeling | Homo sapiens |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| M172L | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme, and altered substrate specificity | Homo sapiens |
| Q307N | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme, and altered substrate specificity | Homo sapiens |
| W345A | site-directed mutagenesis, almost inactive mutant | Homo sapiens |
| W345M | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme, and altered substrate specificity | Homo sapiens |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | kinetics of wild-type and mutant enzymes, overview | Homo sapiens | |
| 0.12 | - |
4-nitrophenyl-beta-D-fucopyranoside | recombinant wild-type enzyme | Homo sapiens |  |
| 0.19 | - |
4-nitrophenyl-alpha-L-arabino-pyranoside | recombinant wild-type enzyme | Homo sapiens | |
| 0.38 | - |
4-nitrophenyl-beta-D-glucopyranoside | recombinant wild-type enzyme | Homo sapiens | |
| 0.9 | - |
4-nitrophenyl-beta-D-galactopyranoside | recombinant wild-type enzyme | Homo sapiens | |
| 5.1 | - |
4-nitrophenyl beta-L-arabinopyranoside | recombinant wild-type enzyme | Homo sapiens | |
| 18.4 | - |
apigenin-7-O-beta-D-glucoside | recombinant wild-type enzyme | Homo sapiens | |
| 30.2 | - |
quercetin-4'-O-beta-D-glucoside | recombinant wild-type enzyme | Homo sapiens | |
| 36.1 | - |
quercetin-7-O-beta-D-glucoside | recombinant wild-type enzyme | Homo sapiens | |
| 122 | - |
eriodictyol-7-glucoside | recombinant wild-type enzyme | Homo sapiens | |
| 148 | - |
naringenin-7-glucoside | recombinant wild-type enzyme | Homo sapiens | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| cytosol | - |
Homo sapiens | 5829 | - |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Homo sapiens | Q9H227 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| native enzyme 12fold in a two-step process involving hydrophobic interaction chromatography, with an additional anion exchange chromatography step | Homo sapiens |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| celloheptaose + 6 H2O = 7 beta-D-glucose | the two catalytic glutamate residues are located on strand 4, the acid/base Glu165, and on strand 7,the nucleophile Glu373 | Homo sapiens |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
substrate specificity of wild-type and mutant enzymes, overview | Homo sapiens |
| 2.1 | - |
purified enzyme, substrate 4-nitrophenyl-alpha-L-arabinoopyranoside, recombinant wild-type enzyme | Homo sapiens |
| 3.5 | - |
purified enzyme, substrate 4-nitrophenyl-beta-D-glucopyranoside, recombinant wild-type enzyme | Homo sapiens |
| 3.6 | - |
purified enzyme, substrate 4-nitrophenyl-beta-D-fucopyranoside, recombinant wild-type enzyme | Homo sapiens |
| 5.7 | - |
purified enzyme, substrate 4-nitrophenyl-beta-D-galactopyranoside, recombinant wild-type enzyme | Homo sapiens |
| 41.8 | - |
purified enzyme, substrate apigenin-7-glucoside, recombinant wild-type enzyme | Homo sapiens |
| 49.7 | - |
purified enzyme, substrate quercetin-4'-glucoside, recombinant wild-type enzyme | Homo sapiens |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-nitrophenyl beta-D-fucopyranoside + H2O | - |
Homo sapiens | 4-nitrophenol + beta-D-fucose | - |
? | |
| 4-nitrophenyl beta-D-galactopyranoside + H2O | - |
Homo sapiens | 4-nitrophenol + beta-D-galactose | - |
? | |
| 4-nitrophenyl beta-D-glucopyranoside + H2O | - |
Homo sapiens | 4-nitrophenol + beta-D-glucose | - |
? | |
| 4-nitrophenyl beta-L-arabinopyranoside + H2O | - |
Homo sapiens | 4-nitrophenol + beta-L-arabinopyranose | - |
? | |
| 4-nitrophenyl-alpha-L-arabinopyranoside + H2O | - |
Homo sapiens | 4-nitrophenol + alpha-L-arabinose | - |
? | |
| apigenin-7-O-beta-D-glucoside + H2O | high activity | Homo sapiens | apigenin + beta-D-glucose | - |
? | |
| additional information | the human cytosolic beta-glucosidase is a xenobiotic-metabolizing enzyme that hydrolyses certain flavonoid glucosides, with specificity depending on the aglycone moiety, the type of sugar and the linkage between them, substrate specificity, overview | Homo sapiens | ? | - |
? | |
| quercetin 4'-O-beta-D-glucopyranoside + H2O | best substrate, homology modelling revealed that hydrophobic interactions dominate the binding of the aglycone moiety, Val168, Trp345, Phe225, Phe179, Phe334 and Phe433 are especially important in determining substrate specificity | Homo sapiens | quercetin + beta-D-glucose | - |
? | |
| quercetin 7-O-beta-D-glucopyranoside + H2O | high activity | Homo sapiens | quercetin + beta-D-glucose | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| More | the main-chain fold of the enzyme belongs to the (beta/alpha)8 barrel structure, the active site is located at the bottom of a pocket formed by large surface loops, surrounding the C termini of the barrel of beta-strands | Homo sapiens |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| More | the enzyme belongs to the glycosyl hydrolase family 1 | Homo sapiens |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.29 | - |
4-nitrophenyl beta-L-arabinopyranoside | recombinant wild-type enzyme | Homo sapiens | |
| 0.6 | - |
quercetin-7-O-beta-D-glucoside | recombinant wild-type enzyme | Homo sapiens | |
| 0.73 | - |
eriodictyol-7-glucoside | recombinant wild-type enzyme | Homo sapiens | |
| 0.77 | - |
apigenin-7-O-beta-D-glucoside | recombinant wild-type enzyme | Homo sapiens | |
| 1.4 | - |
naringenin-7-glucoside | recombinant wild-type enzyme | Homo sapiens | |
| 1.5 | - |
quercetin-4'-O-beta-D-glucoside | recombinant wild-type enzyme | Homo sapiens | |
| 1.88 | - |
4-nitrophenyl-alpha-L-arabino-pyranoside | recombinant wild-type enzyme | Homo sapiens | |
| 3.12 | - |
4-nitrophenyl-beta-D-glucopyranoside | recombinant wild-type enzyme | Homo sapiens | |
| 3.13 | - |
4-nitrophenyl-beta-D-fucopyranoside | recombinant wild-type enzyme | Homo sapiens | |
| 5.53 | - |
4-nitrophenyl-beta-D-galactopyranoside | recombinant wild-type enzyme | Homo sapiens | |
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