Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli MV1184 cells | Bacteroides thetaiotaomicron |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapor diffusion method, using 100 mM MES-NaOH buffer, pH 6.2, containing 12% (w/v) PEG (polyethylene glycol) 6000 or 100 mM MES-NaOH buffer containing 10% (w/v) PEG 10000, at 20°C | Bacteroides thetaiotaomicron |
Protein Variants | Comment | Organism |
---|---|---|
D415G | inactive | Bacteroides thetaiotaomicron |
D415N | the catalytic activity of the mutant enzyme E470Q is significantly decreased (kcat is 100000 lower than that of the wild type enzyme whereas the Km values of both D415N and wild type enzymes are roughly similar.) | Bacteroides thetaiotaomicron |
E470Q | the catalytic activity of the mutant enzyme E470Q is significantly decreased (kcat is 28000 lower than that of the wild type enzyme whereas the Km values of both D415N and wild type enzymes are the same) | Bacteroides thetaiotaomicron |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.19 | - |
4-nitrophenyl alpha-D-galactoside | mutant enzyme D415N, in 40 mM sodium phosphate buffer, pH 6.0, at 35°C | Bacteroides thetaiotaomicron | |
0.31 | - |
4-nitrophenyl alpha-D-galactoside | wild type enzyme, in 40 mM sodium phosphate buffer, pH 6.0, at 35°C | Bacteroides thetaiotaomicron | |
0.32 | - |
4-nitrophenyl alpha-D-galactoside | mutant enzyme E470Q, in 40 mM sodium phosphate buffer, pH 6.0, at 35°C | Bacteroides thetaiotaomicron |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacteroides thetaiotaomicron | Q8A6L0 | strain ATCC 29148 | - |
Purification (Comment) | Organism |
---|---|
Ni-chelating Sepharose column chromatography and DEAE-Sepharose column chromatography | Bacteroides thetaiotaomicron |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1-azido-1-deoxy-beta-D-galactopyranoside + H2O | - |
Bacteroides thetaiotaomicron | ? | - |
? | |
4-nitrophenyl alpha-D-galactoside + H2O | - |
Bacteroides thetaiotaomicron | 4-nitrophenol + alpha-D-galactose | - |
? |
Synonyms | Comment | Organism |
---|---|---|
GH97b | - |
Bacteroides thetaiotaomicron |
retaining alpha-galactosidase | - |
Bacteroides thetaiotaomicron |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00218 | - |
4-nitrophenyl alpha-D-galactoside | mutant enzyme D415N, in 40 mM sodium phosphate buffer, pH 6.0, at 35°C | Bacteroides thetaiotaomicron | |
0.0684 | - |
4-nitrophenyl alpha-D-galactoside | mutant enzyme E470Q, in 40 mM sodium phosphate buffer, pH 6.0, at 35°C | Bacteroides thetaiotaomicron | |
244 | - |
4-nitrophenyl alpha-D-galactoside | wild type enzyme, in 40 mM sodium phosphate buffer, pH 6.0, at 35°C | Bacteroides thetaiotaomicron |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0115 | - |
4-nitrophenyl alpha-D-galactoside | mutant enzyme D415N, in 40 mM sodium phosphate buffer, pH 6.0, at 35°C | Bacteroides thetaiotaomicron | |
0.213 | - |
4-nitrophenyl alpha-D-galactoside | mutant enzyme E470Q, in 40 mM sodium phosphate buffer, pH 6.0, at 35°C | Bacteroides thetaiotaomicron | |
778 | - |
4-nitrophenyl alpha-D-galactoside | wild type enzyme, in 40 mM sodium phosphate buffer, pH 6.0, at 35°C | Bacteroides thetaiotaomicron |