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Literature summary extracted from

  • Tribolo, S.; Berrin, J.G.; Kroon, P.A.; Czjzek, M.; Juge, N.
    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.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.21 expression of wild-type and mutant enzymes in Pichia pastoris Homo sapiens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.21 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

EC Number Protein Variants Comment Organism
3.2.1.21 M172L site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme, and altered substrate specificity Homo sapiens
3.2.1.21 Q307N site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme, and altered substrate specificity Homo sapiens
3.2.1.21 W345A site-directed mutagenesis, almost inactive mutant Homo sapiens
3.2.1.21 W345M site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme, and altered substrate specificity Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.21 additional information
-
 additional information kinetics of wild-type and mutant enzymes, overview Homo sapiens
3.2.1.21 0.12
-
 4-nitrophenyl-beta-D-fucopyranoside recombinant wild-type enzyme Homo sapiens
3.2.1.21 0.19
-
 4-nitrophenyl-alpha-L-arabino-pyranoside recombinant wild-type enzyme Homo sapiens
3.2.1.21 0.38
-
 4-nitrophenyl-beta-D-glucopyranoside recombinant wild-type enzyme Homo sapiens
3.2.1.21 0.9
-
 4-nitrophenyl-beta-D-galactopyranoside recombinant wild-type enzyme Homo sapiens
3.2.1.21 5.1
-
 4-nitrophenyl beta-L-arabinopyranoside recombinant wild-type enzyme Homo sapiens
3.2.1.21 18.4
-
 apigenin-7-O-beta-D-glucoside recombinant wild-type enzyme Homo sapiens
3.2.1.21 30.2
-
 quercetin-4'-O-beta-D-glucoside recombinant wild-type enzyme Homo sapiens
3.2.1.21 36.1
-
 quercetin-7-O-beta-D-glucoside recombinant wild-type enzyme Homo sapiens
3.2.1.21 122
-
 eriodictyol-7-glucoside recombinant wild-type enzyme Homo sapiens
3.2.1.21 148
-
 naringenin-7-glucoside recombinant wild-type enzyme Homo sapiens

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.2.1.21 cytosol
-
 Homo sapiens 5829
-

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.21 Homo sapiens Q9H227
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.21 native enzyme 12fold in a two-step process involving hydrophobic interaction chromatography, with an additional anion exchange chromatography step Homo sapiens

Reaction

EC Number Reaction Comment Organism Reaction ID
3.2.1.21 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
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.21 additional information
-
 substrate specificity of wild-type and mutant enzymes, overview Homo sapiens
3.2.1.21 2.1
-
 purified enzyme, substrate 4-nitrophenyl-alpha-L-arabinoopyranoside, recombinant wild-type enzyme Homo sapiens
3.2.1.21 3.5
-
 purified enzyme, substrate 4-nitrophenyl-beta-D-glucopyranoside, recombinant wild-type enzyme Homo sapiens
3.2.1.21 3.6
-
 purified enzyme, substrate 4-nitrophenyl-beta-D-fucopyranoside, recombinant wild-type enzyme Homo sapiens
3.2.1.21 5.7
-
 purified enzyme, substrate 4-nitrophenyl-beta-D-galactopyranoside, recombinant wild-type enzyme Homo sapiens
3.2.1.21 41.8
-
 purified enzyme, substrate apigenin-7-glucoside, recombinant wild-type enzyme Homo sapiens
3.2.1.21 49.7
-
 purified enzyme, substrate quercetin-4'-glucoside, recombinant wild-type enzyme Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.21 4-nitrophenyl beta-D-fucopyranoside + H2O
-
 Homo sapiens 4-nitrophenol + beta-D-fucose
-
 ?
3.2.1.21 4-nitrophenyl beta-D-galactopyranoside + H2O
-
 Homo sapiens 4-nitrophenol + beta-D-galactose
-
 ?
3.2.1.21 4-nitrophenyl beta-D-glucopyranoside + H2O
-
 Homo sapiens 4-nitrophenol + beta-D-glucose
-
 ?
3.2.1.21 4-nitrophenyl beta-L-arabinopyranoside + H2O
-
 Homo sapiens 4-nitrophenol + beta-L-arabinopyranose
-
 ?
3.2.1.21 4-nitrophenyl-alpha-L-arabinopyranoside + H2O
-
 Homo sapiens 4-nitrophenol + alpha-L-arabinose
-
 ?
3.2.1.21 apigenin-7-O-beta-D-glucoside + H2O high activity Homo sapiens apigenin + beta-D-glucose
-
 ?
3.2.1.21 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 ?
-
 ?
3.2.1.21 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
-
 ?
3.2.1.21 quercetin 7-O-beta-D-glucopyranoside + H2O high activity Homo sapiens quercetin + beta-D-glucose
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.21 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

EC Number Synonyms Comment Organism
3.2.1.21 More the enzyme belongs to the glycosyl hydrolase family 1 Homo sapiens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.21 0.29
-
 4-nitrophenyl beta-L-arabinopyranoside recombinant wild-type enzyme Homo sapiens
3.2.1.21 0.6
-
 quercetin-7-O-beta-D-glucoside recombinant wild-type enzyme Homo sapiens
3.2.1.21 0.73
-
 eriodictyol-7-glucoside recombinant wild-type enzyme Homo sapiens
3.2.1.21 0.77
-
 apigenin-7-O-beta-D-glucoside recombinant wild-type enzyme Homo sapiens
3.2.1.21 1.4
-
 naringenin-7-glucoside recombinant wild-type enzyme Homo sapiens
3.2.1.21 1.5
-
 quercetin-4'-O-beta-D-glucoside recombinant wild-type enzyme Homo sapiens
3.2.1.21 1.88
-
 4-nitrophenyl-alpha-L-arabino-pyranoside recombinant wild-type enzyme Homo sapiens
3.2.1.21 3.12
-
 4-nitrophenyl-beta-D-glucopyranoside recombinant wild-type enzyme Homo sapiens
3.2.1.21 3.13
-
 4-nitrophenyl-beta-D-fucopyranoside recombinant wild-type enzyme Homo sapiens
3.2.1.21 5.53
-
 4-nitrophenyl-beta-D-galactopyranoside recombinant wild-type enzyme Homo sapiens