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Literature summary extracted from

  • Hwa, K.Y.; Subramani, B.; Shen, S.T.; Lee, Y.M.
    Exchange of active site residues alters substrate specificity in extremely thermostable beta-glycosidase from Thermococcus kodakarensis KOD1 (2015), Enzyme Microb. Technol., 77, 14-20 .
    View publication on PubMed

Application

EC Number Application Comment Organism
3.2.1.21 degradation the high-catalytic turn-over rate by mutant enzyme D206N for beta-glucosidase activity makes it a useful enzyme in cellulose degradation at high temperatures Thermococcus kodakarensis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.21 wild type and mutant enzymes Q77R, D206N, D206Q, E207S and E399S are expressed in Escherichia coli Thermococcus kodakarensis
3.2.1.25 wild type and mutant enzymes Q77R, D206N, D206Q, E207S and E399S are expressed in Escherichia coli Thermococcus kodakarensis
3.2.1.38 wild type and mutant enzymes Q77R, D206N, D206Q, E207S and E399S are expressed in Escherichia coli Thermococcus kodakarensis

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.21 D206N catalytically active mutant enzyme, similar temperature optimum like wild-type enzyme. The high-catalytic turn-over rate by D206N for beta-glucosidase activity makes it a useful enzyme in cellulose degradation at high temperatures Thermococcus kodakarensis
3.2.1.21 D206Q mutant enzyme shows less than 10% hydrolytic activity than the wild-type toward 4-nitrophenyl glycosides Thermococcus kodakarensis
3.2.1.21 E207S no hydrolytic activity Thermococcus kodakarensis
3.2.1.21 E399S no hydrolytic activity Thermococcus kodakarensis
3.2.1.21 Q77R mutant enzyme shows less than 10% hydrolytic activity than the wild-type toward 4-nitrophenyl glycosides Thermococcus kodakarensis
3.2.1.25 D206N catalytically active mutant enzyme, similar temperature optimum like wild-type enzyme. The high-catalytic turn-over rate by D206N for beta-glucosidase activity makes it a useful enzyme in cellulose degradation at high temperatures Thermococcus kodakarensis
3.2.1.25 D206Q mutant enzyme shows less than 10% hydrolytic activity than the wild-type toward 4-nitrophenyl glycosides Thermococcus kodakarensis
3.2.1.25 E207S no hydrolytic activity Thermococcus kodakarensis
3.2.1.25 E399S no hydrolytic activity Thermococcus kodakarensis
3.2.1.25 Q77R mutant enzyme shows less than 10% hydrolytic activity than the wild-type toward 4-nitrophenyl glycosides Thermococcus kodakarensis
3.2.1.38 D206N catalytically active mutant enzyme, similar temperature optimum like wild-type enzyme. The high-catalytic turn-over rate by D206N for beta-glucosidase activity makes it a useful enzyme in cellulose degradation at high temperatures Thermococcus kodakarensis
3.2.1.38 D206Q mutant enzyme shows less than 10% hydrolytic activity than the wild-type toward 4-nitrophenyl glycosides Thermococcus kodakarensis
3.2.1.38 E207S no hydrolytic activity Thermococcus kodakarensis
3.2.1.38 E399S no hydrolytic activity Thermococcus kodakarensis
3.2.1.38 Q77R mutant enzyme shows less than 10% hydrolytic activity than the wild-type toward 4-nitrophenyl glycosides Thermococcus kodakarensis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.21 0.4
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.21 0.94
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.21 1.08
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.21 1.29
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.21 4.6
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.21 9.23
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.25 0.4
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.25 0.94
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.25 1.08
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.25 1.29
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.25 4.6
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.25 9.23
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.38 0.4
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.38 0.94
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.38 1.08
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.38 1.29
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.38 4.6
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.38 9.23
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.21 Thermococcus kodakarensis Q9YGB8
-
-
3.2.1.25 Thermococcus kodakarensis Q9YGB8
-
-
3.2.1.38 Thermococcus kodakarensis Q9YGB8
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.21
-
 Thermococcus kodakarensis
3.2.1.25
-
 Thermococcus kodakarensis
3.2.1.38
-
 Thermococcus kodakarensis

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
-
 Thermococcus kodakarensis 4-nitrophenol + beta-D-fucopyranose
-
 ?
3.2.1.21 4-nitrophenyl beta-D-glucopyranoside + H2O
-
 Thermococcus kodakarensis 4-nitrophenol + beta-D-glucopyranose
-
 ?
3.2.1.21 4-nitrophenyl beta-D-mannopyranoside + H2O
-
 Thermococcus kodakarensis 4-nitrophenol + beta-D-mannopyranose
-
 ?
3.2.1.25 4-nitrophenyl beta-D-fucopyranoside + H2O
-
 Thermococcus kodakarensis 4-nitrophenol + beta-D-fucopyranose
-
 ?
3.2.1.25 4-nitrophenyl beta-D-glucopyranoside + H2O
-
 Thermococcus kodakarensis 4-nitrophenol + beta-D-glucopyranose
-
 ?
3.2.1.25 4-nitrophenyl beta-D-mannopyranoside + H2O
-
 Thermococcus kodakarensis 4-nitrophenol + beta-D-mannopyranose
-
 ?
3.2.1.38 4-nitrophenyl beta-D-fucopyranoside + H2O
-
 Thermococcus kodakarensis 4-nitrophenol + beta-D-fucopyranose
-
 ?
3.2.1.38 4-nitrophenyl beta-D-glucopyranoside + H2O
-
 Thermococcus kodakarensis 4-nitrophenol + beta-D-glucopyranose
-
 ?
3.2.1.38 4-nitrophenyl beta-D-mannopyranoside + H2O
-
 Thermococcus kodakarensis 4-nitrophenol + beta-D-mannopyranose
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.21 20.69
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.21 50.99
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.21 227.64
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.21 322.25
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.21 323.41
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.21 349.62
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.25 20.69
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.25 50.99
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.25 227.64
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.25 322.25
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.25 323.41
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.25 349.62
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.38 20.69
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.38 50.99
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.38 227.64
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.38 322.25
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.38 323.41
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.38 349.62
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis

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 36.9
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.21 47.21
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.21 49.48
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.21 51.72
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.21 250.7
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.21 342.81
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.25 36.9
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.25 47.21
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.25 49.48
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.25 51.72
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.25 250.7
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.25 342.81
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.38 36.9
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.38 47.21
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.38 49.48
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis
3.2.1.38 51.72
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.38 250.7
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH is not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.38 342.81
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH is not specified in the publication, wild-type Thermococcus kodakarensis