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Literature summary for 3.2.1.21 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
Search for more literature for 3.2.1.21

Cloned(Commentary)

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

Protein Variants

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

Inhibitors

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

KM Value [mM]

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

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

Organism

Organism UniProt Comment Textmining
Aspergillus niger
-
-
-
Aspergillus niger ASKU28
-
-
-

Substrates and Products (Substrate)

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

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
4
-
-
 Aspergillus niger

kcat/KM [mM/s]

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