Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Lv, B.; Sun, H.; Huang, S.; Feng, X.; Jiang, T.; Li, C.
    Structure-guided engineering of the substrate specificity of a fungal beta-glucuronidase toward triterpenoid saponins (2018), J. Biol. Chem., 293, 433-443 .
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
3.2.1.31 synthesis mutant enzyme A365H/R563E has great potential in the industrial production of glycyrrhetinic acid 3-O-mono-beta-D-glucuronide Aspergillus oryzae

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.31 expression of the protein His6-tagged at its N terminus in Escherichia coli Aspergillus oryzae

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.31 sitting drop vapor diffusion method at 16°C, structure of the enzyme is determined at 3.1 A resolution and structure of the inactive mutant enzyme E414D/E505D in complex with the native substrate glycyrrhetic acid 3-O-mono-beta-glucuronide at 2.6 A resolution Aspergillus oryzae

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.31 A365H/R563E the yield of glycyrrhetinic acid 3-O-mono-beta-D-glucuronide of the mutant enzyme is 96% compared to less than 10% produced by wild-type enzyme, resulting in nearly complete alteration of the substrate selectivity of the glucuronyl hydrolase from glycyrrhetic acid to glycyrrhetinic acid 3-O-mono-beta-D-glucuronide formation. No activity with glycyrrhetinic acid 3-O-mono-beta-D-glucuronide Aspergillus oryzae
3.2.1.31 A365Q the yield of glycyrrhetinic acid 3-O-mono-beta-D-glucuronide of the mutant enzyme is 65% compared to less than 10% produced by wild-type enzyme Aspergillus oryzae
3.2.1.31 A365T the yield of glycyrrhetinic acid 3-O-mono-beta-D-glucuronide of the mutant enzyme is 61% compared to less than 10% produced by wild-type enzyme Aspergillus oryzae
3.2.1.31 A365T/R563E the yield of glycyrrhetinic acid 3-O-mono-beta-D-glucuronide of the mutant enzyme is 95% compared to less than 10% produced by wild-type enzyme, resulting in nearly complete alteration of the substrate selectivity of the glucuronyl hydrolase from glycyrrhetic acid to glycyrrhetinic acid 3-O-mono-beta-D-glucuronide formation. No activity with glycyrrhetinic acid 3-O-mono-beta-D-glucuronide Aspergillus oryzae
3.2.1.31 E414A inactive mutant enzyme Aspergillus oryzae
3.2.1.31 E414D/E505D inactive mutant enzyme Aspergillus oryzae
3.2.1.31 E505A inactive mutant enzyme Aspergillus oryzae
3.2.1.31 R563E the yield of glycyrrhetinic acid 3-O-mono-beta-D-glucuronide of the mutant enzyme is 77% compared to less than 10% produced by wild-type enzyme Aspergillus oryzae
3.2.1.31 R563K the yield of glycyrrhetinic acid 3-O-mono-beta-D-glucuronide of the mutant enzyme is 58% compared to less than 10% produced by wild-type enzyme. The catalytic efficiency (kcat/Km) toward glycyrrhetinic acid 3-O-mono-beta-D-glucuronide decreases by 88.4%, whereas kcat/Km toward GL decreases by 12.3%, confirming site 563 has a dramatic effect on substrate specificity Aspergillus oryzae
3.2.1.31 R563Q the yield of glycyrrhetinic acid 3-O-mono-beta-D-glucuronide of the mutant enzyme is 42% compared to less than 10% produced by wild-type enzyme Aspergillus oryzae
3.2.1.31 V447Q the yield of glycyrrhetinic acid 3-O-mono-beta-D-glucuronide of the mutant enzyme is 81% compared to less than 10% produced by wild-type enzyme Aspergillus oryzae
3.2.1.31 V447Q/R563K the yield of glycyrrhetinic acid 3-O-mono-beta-D-glucuronide of the mutant enzyme is 95% compared to less than 10% produced by wild-type enzyme, resulting in nearly complete alteration of the substrate selectivity of the glucuronyl hydrolase from glycyrrhetic acid to glycyrrhetinic acid 3-O-mono-beta-D-glucuronide formation. No activity with glycyrrhetinic acid 3-O-mono-beta-D-glucuronide Aspergillus oryzae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.31 0.78
-
 glycyrrhizin pH 5.0, 40°C, wild-type enzyme Aspergillus oryzae
3.2.1.31 0.91
-
 glycyrrhetinic acid 3-O-mono-beta-D-glucuronide pH 5.0, 40°C, mutant enzyme R563K Aspergillus oryzae
3.2.1.31 0.92
-
 glycyrrhetinic acid 3-O-mono-beta-D-glucuronide pH 5.0, 40°C, wild-type enzyme Aspergillus oryzae
3.2.1.31 0.97
-
 glycyrrhizin pH 5.0, 40°C, mutant enzyme R563K Aspergillus oryzae
3.2.1.31 1.06
-
 glycyrrhetinic acid 3-O-mono-beta-D-glucuronide pH 5.0, 40°C, mutant enzyme V447Q Aspergillus oryzae
3.2.1.31 1.1
-
 glycyrrhetinic acid 3-O-mono-beta-D-glucuronide pH 5.0, 40°C, mutant enzyme A365T Aspergillus oryzae
3.2.1.31 1.21
-
 glycyrrhizin pH 5.0, 40°C, mutant enzyme V447Q Aspergillus oryzae
3.2.1.31 1.23
-
 glycyrrhizin pH 5.0, 40°C, mutant enzyme A365T Aspergillus oryzae
3.2.1.31 1.62
-
 glycyrrhizin pH 5.0, 40°C, mutant enzyme V447Q/R563K Aspergillus oryzae
3.2.1.31 2.2
-
 glycyrrhizin pH 5.0, 40°C, mutant enzyme A365T/R563E Aspergillus oryzae
3.2.1.31 2.68
-
 glycyrrhizin pH 5.0, 40°C, mutant enzyme A365H/R563E Aspergillus oryzae

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.31 290000
-
 ultracentrifugation Aspergillus oryzae

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.31 Aspergillus oryzae A7XS03
-
-
3.2.1.31 Aspergillus oryzae Li-3 A7XS03
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.31 Ni2+-affinity chromatography, anion-exchange chromatography, and gel filtration Aspergillus oryzae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.31 4-nitrophenol beta-D-glucuronide + H2O
-
 Aspergillus oryzae 4-nitrophenol + beta-D-glucuronate
-
 ?
3.2.1.31 4-nitrophenol beta-D-glucuronide + H2O
-
 Aspergillus oryzae Li-3 4-nitrophenol + beta-D-glucuronate
-
 ?
3.2.1.31 glycyrrhetinic acid 3-O-mono-beta-D-glucuronide + H2O the enzyme shows similar activities toward both glycyrrhizin and glycyrrhetinic acid 3-O-mono-beta-D-glucuronide Aspergillus oryzae glycyrrhetic acid + D-glucuronate
-
 ?
3.2.1.31 glycyrrhetinic acid 3-O-mono-beta-D-glucuronide + H2O the enzyme shows similar activities toward both glycyrrhizin and glycyrrhetinic acid 3-O-mono-beta-D-glucuronide Aspergillus oryzae Li-3 glycyrrhetic acid + D-glucuronate
-
 ?
3.2.1.31 glycyrrhizin + H2O the enzyme shows similar activities toward both glycyrrhizin and glycyrrhetinic acid 3-O-mono-beta-D-glucuronide Aspergillus oryzae glycyrrhetinic acid 3-O-mono-beta-D-glucuronide + D-glucuronate
-
 ?
3.2.1.31 glycyrrhizin + H2O the enzyme shows similar activities toward both glycyrrhizin and glycyrrhetinic acid 3-O-mono-beta-D-glucuronide Aspergillus oryzae Li-3 glycyrrhetinic acid 3-O-mono-beta-D-glucuronide + D-glucuronate
-
 ?
3.2.1.31 additional information the enzyme does not show strict specificity toward the aglycone moiety and exhibits activity toward all the natural and artificial substrates. Strict glycan specificity and hydrolyzes only the artificial substrate containing glucuronide groups Aspergillus oryzae ?
-
 ?
3.2.1.31 additional information the enzyme does not show strict specificity toward the aglycone moiety and exhibits activity toward all the natural and artificial substrates. Strict glycan specificity and hydrolyzes only the artificial substrate containing glucuronide groups Aspergillus oryzae Li-3 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.31 homotetramer each monomer comprises three distinct domains: a sugar-binding, an immunoglobulin-like beta-sandwich, and a TIM barrel domain Aspergillus oryzae

Synonyms

EC Number Synonyms Comment Organism
3.2.1.31 PGUS
-
 Aspergillus oryzae

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.31 40
-
 assay at Aspergillus oryzae

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.31 0.89
-
 glycyrrhetinic acid 3-O-mono-beta-D-glucuronide pH 5.0, 40°C, mutant enzyme R563K Aspergillus oryzae
3.2.1.31 2.72
-
 glycyrrhizin pH 5.0, 40°C, mutant enzyme A365T/R563E Aspergillus oryzae
3.2.1.31 3.06
-
 glycyrrhizin pH 5.0, 40°C, mutant enzyme A365H/R563E Aspergillus oryzae
3.2.1.31 3.23
-
 glycyrrhizin pH 5.0, 40°C, wild-type enzyme Aspergillus oryzae
3.2.1.31 3.28
-
 glycyrrhizin pH 5.0, 40°C, mutant enzyme A365T Aspergillus oryzae
3.2.1.31 3.32
-
 glycyrrhetinic acid 3-O-mono-beta-D-glucuronide pH 5.0, 40°C, mutant enzyme V447Q Aspergillus oryzae
3.2.1.31 3.52
-
 glycyrrhizin pH 5.0, 40°C, mutant enzyme R563K Aspergillus oryzae
3.2.1.31 4.36
-
 glycyrrhetinic acid 3-O-mono-beta-D-glucuronide pH 5.0, 40°C, mutant enzyme A365T Aspergillus oryzae
3.2.1.31 6.03
-
 glycyrrhizin pH 5.0, 40°C, mutant enzyme V447Q Aspergillus oryzae
3.2.1.31 7.01
-
 glycyrrhizin pH 5.0, 40°C, mutant enzyme V447Q/R563K Aspergillus oryzae
3.2.1.31 7.81
-
 glycyrrhetinic acid 3-O-mono-beta-D-glucuronide pH 5.0, 40°C, wild-type enzyme Aspergillus oryzae

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.31 5
-
 assay at Aspergillus oryzae

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.31 0.98
-
 glycyrrhetinic acid 3-O-mono-beta-D-glucuronide pH 5.0, 40°C, mutant enzyme R563K Aspergillus oryzae
3.2.1.31 1.14
-
 glycyrrhizin pH 5.0, 40°C, mutant enzyme A365H/R563E Aspergillus oryzae
3.2.1.31 1.23
-
 glycyrrhizin pH 5.0, 40°C, mutant enzyme A365T/R563E Aspergillus oryzae
3.2.1.31 2.67
-
 glycyrrhizin pH 5.0, 40°C, mutant enzyme A365T Aspergillus oryzae
3.2.1.31 3.06
-
 glycyrrhetinic acid 3-O-mono-beta-D-glucuronide pH 5.0, 40°C, mutant enzyme A365T Aspergillus oryzae
3.2.1.31 3.13
-
 glycyrrhetinic acid 3-O-mono-beta-D-glucuronide pH 5.0, 40°C, mutant enzyme V447Q Aspergillus oryzae
3.2.1.31 3.63
-
 glycyrrhizin pH 5.0, 40°C, mutant enzyme R563K Aspergillus oryzae
3.2.1.31 4.14
-
 glycyrrhizin pH 5.0, 40°C, wild-type enzyme Aspergillus oryzae
3.2.1.31 4.32
-
 glycyrrhizin pH 5.0, 40°C, mutant enzyme V447Q/R563K Aspergillus oryzae
3.2.1.31 4.98
-
 glycyrrhizin pH 5.0, 40°C, mutant enzyme V447Q Aspergillus oryzae
3.2.1.31 8.49
-
 glycyrrhetinic acid 3-O-mono-beta-D-glucuronide pH 5.0, 40°C, wild-type enzyme Aspergillus oryzae