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Literature summary extracted from

  • Thongpoo, P.; McKee, L.S.; Araujo, A.C.; Kongsaeree, P.T.; Brumer, H.
    Identification of the acid/base catalyst of a glycoside hydrolase family 3 (GH3) beta-glucosidase from Aspergillus niger ASKU28 (2013), Biochim. Biophys. Acta, 1830, 2739-2749.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.21 expressed in Pichia pastoris strain Y11430 Aspergillus niger

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.21 D463A the variant exhibits kinetic constants similar to the wild type enzyme Aspergillus niger
3.2.1.21 D487A the variant exhibits kinetic constants similar to the wild type enzyme Aspergillus niger
3.2.1.21 D496A the variant exhibits kinetic constants similar to the wild type enzyme Aspergillus niger
3.2.1.21 D501A the variant exhibits kinetic constants similar to the wild type enzyme Aspergillus niger
3.2.1.21 E490A the variant exhibits greatly reduced kcat and kcat/Km values toward the natural substrate cellobiose (67000 and 61000fold, respectively) Aspergillus niger

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.21 Sodium azide about 50% reduced activity at 500 mM Aspergillus niger

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.21 0.015
-
2,4-dinitrophenyl beta-D-glucoside mutant enzyme E490A, in 0.1 M sodium acetate, pH 6.0, at 40°C Aspergillus niger
3.2.1.21 0.1
-
2,4-dinitrophenyl beta-D-glucoside mutant enzyme E490A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 0.26
-
2,4-dinitrophenyl beta-D-glucoside wild type enzyme, in 0.1 M sodium acetate, pH 6.0, at 40°C Aspergillus niger
3.2.1.21 0.44
-
4-nitrophenyl beta-D-glucoside mutant enzyme D496A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 0.49
-
2,4-dinitrophenyl beta-D-glucoside wild type enzyme, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 0.54
-
4-nitrophenyl beta-D-glucoside wild type enzyme, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 0.55
-
4-nitrophenyl beta-D-glucoside mutant enzyme D487A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 0.57
-
cellobiose mutant enzyme D496A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 0.64
-
4-nitrophenyl beta-D-glucoside mutant enzyme D463A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 0.97
-
4-nitrophenyl beta-D-glucoside mutant enzyme D501A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 1
-
cellobiose mutant enzyme D463A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 1.02
-
cellobiose mutant enzyme E490A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 1.03
-
cellobiose mutant enzyme D487A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 1.11
-
cellobiose wild type enzyme, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 1.36
-
cellobiose mutant enzyme D501A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 6.09
-
4-nitrophenyl beta-D-glucoside mutant enzyme E490A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.21 cellobiose + H2O Aspergillus niger
-
2 beta-D-glucose
-
?
3.2.1.21 cellobiose + H2O Aspergillus niger ASKU28
-
2 beta-D-glucose
-
?

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.21 Aspergillus niger
-
-
-
3.2.1.21 Aspergillus niger ASKU28
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.21 2,4-dinitrophenyl beta-D-glucoside + H2O
-
Aspergillus niger ?
-
?
3.2.1.21 2,4-dinitrophenyl beta-D-glucoside + H2O
-
Aspergillus niger ASKU28 ?
-
?
3.2.1.21 4-nitrophenyl beta-D-glucoside + H2O
-
Aspergillus niger 4-nitrophenol + beta-D-glucose
-
?
3.2.1.21 4-nitrophenyl beta-D-glucoside + H2O
-
Aspergillus niger ASKU28 4-nitrophenol + beta-D-glucose
-
?
3.2.1.21 cellobiose + H2O
-
Aspergillus niger 2 beta-D-glucose
-
?
3.2.1.21 cellobiose + H2O
-
Aspergillus niger ASKU28 2 beta-D-glucose
-
?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.21 0.001
-
cellobiose mutant enzyme E490A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 0.32
-
4-nitrophenyl beta-D-glucoside mutant enzyme E490A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 0.34
-
2,4-dinitrophenyl beta-D-glucoside mutant enzyme E490A, in 0.1 M sodium acetate, pH 6.0, at 40°C Aspergillus niger
3.2.1.21 2.12
-
2,4-dinitrophenyl beta-D-glucoside mutant enzyme E490A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 47.3
-
cellobiose mutant enzyme D501A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 67.4
-
cellobiose wild type enzyme, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 71
-
2,4-dinitrophenyl beta-D-glucoside wild type enzyme, in 0.1 M sodium acetate, pH 6.0, at 40°C Aspergillus niger
3.2.1.21 78.3
-
cellobiose mutant enzyme D496A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 78.4
-
cellobiose mutant enzyme D463A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 87.7
-
cellobiose mutant enzyme D487A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 104
-
4-nitrophenyl beta-D-glucoside mutant enzyme D501A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 141
-
4-nitrophenyl beta-D-glucoside mutant enzyme D496A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 141
-
4-nitrophenyl beta-D-glucoside wild type enzyme, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 160
-
4-nitrophenyl beta-D-glucoside mutant enzyme D463A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 163
-
4-nitrophenyl beta-D-glucoside mutant enzyme D487A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 422
-
2,4-dinitrophenyl beta-D-glucoside wild type enzyme, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.21 4
-
-
Aspergillus niger

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.21 0.001
-
cellobiose mutant enzyme E490A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 0.05
-
4-nitrophenyl beta-D-glucoside mutant enzyme E490A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 21.2
-
2,4-dinitrophenyl beta-D-glucoside mutant enzyme E490A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 22.6
-
2,4-dinitrophenyl beta-D-glucoside mutant enzyme E490A, in 0.1 M sodium acetate, pH 6.0, at 40°C Aspergillus niger
3.2.1.21 34.7
-
cellobiose mutant enzyme D501A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 60.6
-
cellobiose wild type enzyme, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 78.7
-
cellobiose mutant enzyme D463A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 85.2
-
cellobiose mutant enzyme D487A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 107
-
4-nitrophenyl beta-D-glucoside mutant enzyme D501A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 138
-
cellobiose mutant enzyme D496A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 249
-
4-nitrophenyl beta-D-glucoside mutant enzyme D463A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 263
-
4-nitrophenyl beta-D-glucoside wild type enzyme, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 273
-
2,4-dinitrophenyl beta-D-glucoside wild type enzyme, in 0.1 M sodium acetate, pH 6.0, at 40°C Aspergillus niger
3.2.1.21 297
-
4-nitrophenyl beta-D-glucoside mutant enzyme D487A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 321
-
4-nitrophenyl beta-D-glucoside mutant enzyme D496A, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger
3.2.1.21 863
-
2,4-dinitrophenyl beta-D-glucoside wild type enzyme, in 0.1 M sodium acetate, pH 4.0, at 40°C Aspergillus niger