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Literature summary extracted from

  • Wu, Y.; Yuan, S.; Chen, S.; Wu, D.; Chen, J.; Wu, J.
    Enhancing the production of galacto-oligosaccharides by mutagenesis of Sulfolobus solfataricus beta-galactosidase (2013), Food Chem., 138, 1588-1595.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.23 expressed in Escherichia coli BL21(DE3) Saccharolobus solfataricus

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.23 F359Q compared to wild-type enzyme the mutant enzyme is less stable at 75°C, the optimum is similar to that of wild-type enzyme. Km of the mutant enzyme is increased about 2fold and the kcat is increased by 32.3% Saccharolobus solfataricus
3.2.1.23 F441Y compared to wild-type enzyme the mutant enzyme is less stable at 75°C, the optimum is similar to that of wild-type enzyme. Km of the mutant enzyme is increased about 2fold and the kcat is increased by 36.6% Saccharolobus solfataricus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.23 14.2
-
 lactose 30°C, pH 6.5, wild-type enzyme Saccharolobus solfataricus
3.2.1.23 29.4
-
 lactose 30°C, pH 6.5, mutant enzyme F441Y Saccharolobus solfataricus
3.2.1.23 31.8
-
 lactose 30°C, pH 6.5, mutant enzyme F359Q Saccharolobus solfataricus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.23 56561
-
 x * 56561, calculated from sequence Saccharolobus solfataricus
3.2.1.23 56600
-
 x * 56600, SDS-PAGE Saccharolobus solfataricus

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.23 Saccharolobus solfataricus P22498
-
-
3.2.1.23 Saccharolobus solfataricus P2 P22498
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.23
-
 Saccharolobus solfataricus

Storage Stability

EC Number Storage Stability Organism
3.2.1.23 25°C, pH 6.5, more than 60% activity after 45 days Saccharolobus solfataricus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.23 2-nitrophenyl beta-D-galactopyranose + H2O
-
 Saccharolobus solfataricus 2-nitrophenol + D-galactose
-
 ?
3.2.1.23 2-nitrophenyl beta-D-galactopyranose + H2O
-
 Saccharolobus solfataricus P2 2-nitrophenol + D-galactose
-
 ?
3.2.1.23 lactose + H2O
-
 Saccharolobus solfataricus D-glucose + D-galactose
-
 ?
3.2.1.23 lactose + H2O
-
 Saccharolobus solfataricus P2 D-glucose + D-galactose
-
 ?
3.2.1.23 additional information quantitative determination of the products of the transglycosylation reactions shows that the amount of glucose is always much higher than that of galactose, which indicates that, during the reaction, galactoside of lactose is used in the transglycosylation reaction to form galactooligosaccharides. Compared to the wild-type enzyme, the yields of galacto-oligosaccharides produced by F441Y and F359Q are increased by 10.8% and 7.4%, respectively Saccharolobus solfataricus ?
-
 ?
3.2.1.23 additional information quantitative determination of the products of the transglycosylation reactions shows that the amount of glucose is always much higher than that of galactose, which indicates that, during the reaction, galactoside of lactose is used in the transglycosylation reaction to form galactooligosaccharides. Compared to the wild-type enzyme, the yields of galacto-oligosaccharides produced by F441Y and F359Q are increased by 10.8% and 7.4%, respectively Saccharolobus solfataricus P2 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.23 ? x * 56561, calculated from sequence Saccharolobus solfataricus
3.2.1.23 ? x * 56600, SDS-PAGE Saccharolobus solfataricus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.23 85
-
 substrate: 2-nitrophenyl beta-D-galactopyranoside, at pH 6.5, mutant enzymes F441Y and F359Q Saccharolobus solfataricus
3.2.1.23 90
-
 substrate: 2-nitrophenyl beta-D-galactopyranoside, at pH 6.5, wild-type enzyme Saccharolobus solfataricus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.2.1.23 70 95 70°C: about 60% of maximal activity, 95°C: about 70% of maximal activity Saccharolobus solfataricus
3.2.1.23 80 90 substrate: 2-nitrophenyl beta-D-galactopyranoside, mutant enzymes F441Y and F359Q show 80% of their maximal activity Saccharolobus solfataricus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.23 75
-
 half-life of mutant enzyme F441Y is 75 h, half-life of mutant enzyme F359Q is 85 h. Activity of wild-type enzyme increases slowly within the first 45 h, it retains 90% of its initial activity after incubation for approximately 80 h. After 80 h, the activity decreases rapidly Saccharolobus solfataricus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.23 122.6
-
 lactose 30°C, pH 6.5, wild-type enzyme Saccharolobus solfataricus
3.2.1.23 162.2
-
 lactose 30°C, pH 6.5, mutant enzyme F441Y Saccharolobus solfataricus
3.2.1.23 167.5
-
 lactose 30°C, pH 6.5, mutant enzyme F359Q Saccharolobus solfataricus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.23 6
-
 substrate: 2-nitrophenyl beta-D-galactopyranoside, mutant enzymes F441Y and F359Q Saccharolobus solfataricus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.23 5 6 wild-type enzyme and mutant enzymes F441Y and F359Q Saccharolobus solfataricus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.23 5.2
-
 lactose 30°C, pH 6.5, mutant enzyme F359Q Saccharolobus solfataricus
3.2.1.23 5.5
-
 lactose 30°C, pH 6.5, mutant enzyme F441Y Saccharolobus solfataricus
3.2.1.23 8.6
-
 lactose 30°C, pH 6.5, wild-type enzyme Saccharolobus solfataricus