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Literature summary for 3.2.1.23 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
Search for more literature for 3.2.1.23

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
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
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]

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

Molecular Weight [Da]

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

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus P22498
-
-
Saccharolobus solfataricus P2 P22498
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Saccharolobus solfataricus

Storage Stability

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-nitrophenyl beta-D-galactopyranose + H2O
-
 Saccharolobus solfataricus 2-nitrophenol + D-galactose
-
 ?
2-nitrophenyl beta-D-galactopyranose + H2O
-
 Saccharolobus solfataricus P2 2-nitrophenol + D-galactose
-
 ?
lactose + H2O
-
 Saccharolobus solfataricus D-glucose + D-galactose
-
 ?
lactose + H2O
-
 Saccharolobus solfataricus P2 D-glucose + D-galactose
-
 ?
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 ?
-
 ?
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

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

Temperature Optimum [°C]

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

Temperature Range [°C]

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

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
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]

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

pH Optimum

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

pH Range

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

kcat/KM [mM/s]

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