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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

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.B35 expression in Escherichia coli Thermococcus kodakarensis

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.B35 D206N mutation alters the catalytic turn-over rate for glucosidase and mannosidase activities with fucosidase activity remain unchanged Thermococcus kodakarensis
3.2.1.B35 D206Q catalytically inactive mutant enzyme. The extended side chain of D206Q is predicted to affect the substrate binding during catalysis Thermococcus kodakarensis
3.2.1.B35 E207S catalytically inactive mutant enzyme Thermococcus kodakarensis
3.2.1.B35 E399S catalytically inactive mutant enzyme Thermococcus kodakarensis
3.2.1.B35 Q77R catalytically inactive mutant enzyme. Q77R might have made some changes in three dimensional structure due to its electrostatic effect and lost its catalytic activity Thermococcus kodakarensis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.B35 0.4
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.B35 0.94
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH not specified in the publication, wild-type enzyme Thermococcus kodakarensis
3.2.1.B35 1.08
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH not specified in the publication, wild-type enzyme Thermococcus kodakarensis
3.2.1.B35 1.29
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.B35 4.6
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH not specified in the publication, wild-type enzyme Thermococcus kodakarensis
3.2.1.B35 9.23
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.B35 Thermococcus kodakarensis Q9YGB8
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.B35
-
 Thermococcus kodakarensis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.B35 4-nitrophenyl beta-D-fucopyranoside + H2O
-
 Thermococcus kodakarensis 4-nitrophenol + beta-D-fucopyranose
-
 ?
3.2.1.B35 4-nitrophenyl beta-D-glucopyranoside + H2O
-
 Thermococcus kodakarensis 4-nitrophenol + D-glucopyranose
-
 ?
3.2.1.B35 4-nitrophenyl beta-D-mannopyranoside + H2O
-
 Thermococcus kodakarensis 4-nitrophenol + beta-D-mannopyranose
-
 ?
3.2.1.B35 additional information the enzyme shows beta-glucosidase, beta-mannosidase, beta-fucosidase and beta-galactosidase activities Thermococcus kodakarensis ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.B35 dimer wild-type and mutant enzymes (Q77R, D206N, D206Q, E207S and E399S) Thermococcus kodakarensis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.B35 95 100
-
 Thermococcus kodakarensis

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.2.1.B35 60 100 60°C: about 40% of maximal activity, 95-100°C: optimum Thermococcus kodakarensis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.B35 20.69
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.B35 50.99
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH not specified in the publication, wild-type enzyme Thermococcus kodakarensis
3.2.1.B35 227.6
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH not specified in the publication, wild-type enzyme Thermococcus kodakarensis
3.2.1.B35 322.3
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH not specified in the publication, wild-type enzyme Thermococcus kodakarensis
3.2.1.B35 323.4
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.B35 340.6
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH 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.B35 36.9
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.B35 47.2
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH not specified in the publication, wild-type enzyme Thermococcus kodakarensis
3.2.1.B35 49.48
-
 4-nitrophenyl beta-D-glucopyranoside 80°C, pH not specified in the publication, wild-type enzyme Thermococcus kodakarensis
3.2.1.B35 51.7
-
 4-nitrophenyl beta-D-mannopyranoside 80°C, pH not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.B35 250.7
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH not specified in the publication, mutant enzyme D206N Thermococcus kodakarensis
3.2.1.B35 342.8
-
 4-nitrophenyl beta-D-fucopyranoside 80°C, pH not specified in the publication, wild-type enzyme Thermococcus kodakarensis