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Literature summary extracted from
- The molecular mechanism of thermostable beta-galactosidases AgaA and AgaB explained by X-ray crystallography and mutational studies (2012), J. Biol. Chem., 287, 39642-39652.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 3.2.1.22 | expressed in Escherichia coli RM448 cells | Geobacillus stearothermophilus |
Crystallization (Commentary)
| EC Number | Crystallization (Comment) | Organism |
|---|---|---|
| 3.2.1.22 | vapor diffusion method, using 80 mM sodium acetate, 20% (w/v) polyethylene glycol 4000, and 6.4% (v/v) isopropanol | Geobacillus stearothermophilus |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 3.2.1.22 | W336A | the mutant shows about 28fold reduced catalytic efficiency compared to the wild type | Geobacillus stearothermophilus |
| 3.2.1.22 | W336D | the mutant shows severely reduced catalytic efficiency compared to the wild type | Geobacillus stearothermophilus |
| 3.2.1.22 | W336F | the mutant shows about 3fold increased catalytic efficiency compared to the wild type | Geobacillus stearothermophilus |
| 3.2.1.22 | W336N | the mutant shows about 10fold reduced catalytic efficiency compared to the wild type | Geobacillus stearothermophilus |
| 3.2.1.22 | W336S | the mutant shows about 100fold reduced catalytic efficiency compared to the wild type | Geobacillus stearothermophilus |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.2.1.22 | 0.21 | - |
4-nitrophenyl alpha-D-galactopyranoside | wild type enzyme isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus |  |
| 3.2.1.22 | 0.24 | - |
4-nitrophenyl alpha-D-galactopyranoside | mutant enzyme W336F of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 0.44 | - |
4-nitrophenyl alpha-D-galactopyranoside | mutant enzyme W336A of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 0.83 | - |
4-nitrophenyl alpha-D-galactopyranoside | wild type enzyme isoform AgaB, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 0.84 | - |
4-nitrophenyl alpha-D-galactopyranoside | mutant enzyme W336N of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 1.33 | - |
4-nitrophenyl alpha-D-galactopyranoside | mutant enzyme W336D of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 3.8 | - |
raffinose | wild type enzyme isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 8.7 | - |
4-nitrophenyl alpha-D-galactopyranoside | mutant enzyme W336S of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 36.6 | - |
raffinose | mutant enzyme W336S of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 200 | - |
raffinose | wild type enzyme isoform AgaB, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 205 | - |
raffinose | mutant enzyme W336N of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 250 | - |
raffinose | mutant enzyme W336F of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 252 | - |
raffinose | mutant enzyme W336A of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.2.1.22 | Geobacillus stearothermophilus | Q9ALJ4 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 3.2.1.22 | DMAE column chromatography, Mono Q column chromatography and Superdex 200 gel filtration | Geobacillus stearothermophilus |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.2.1.22 | 4-nitrophenyl alpha-D-galactopyranoside + H2O | - |
Geobacillus stearothermophilus | 4-nitrophenol + alpha-D-galactopyranose | - |
? | |
| 3.2.1.22 | raffinose + H2O | - |
Geobacillus stearothermophilus | ? | - |
? | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 3.2.1.22 | AgaA | isoform | Geobacillus stearothermophilus |
| 3.2.1.22 | AgaB | isoform | Geobacillus stearothermophilus |
| 3.2.1.22 | Alpha-D-galactoside galactohydrolase | - |
Geobacillus stearothermophilus |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.2.1.22 | 0.97 | - |
4-nitrophenyl alpha-D-galactopyranoside | mutant enzyme W336D of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 5.2 | - |
raffinose | mutant enzyme W336S of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 7.9 | - |
4-nitrophenyl alpha-D-galactopyranoside | mutant enzyme W336A of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 10.3 | - |
raffinose | mutant enzyme W336N of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 38.2 | - |
4-nitrophenyl alpha-D-galactopyranoside | mutant enzyme W336N of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 48 | - |
4-nitrophenyl alpha-D-galactopyranoside | mutant enzyme W336S of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 48.1 | - |
raffinose | mutant enzyme W336A of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 105.6 | - |
4-nitrophenyl alpha-D-galactopyranoside | wild type enzyme isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 180 | - |
raffinose | wild type enzyme isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 375 | - |
4-nitrophenyl alpha-D-galactopyranoside | mutant enzyme W336F of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 832 | - |
raffinose | mutant enzyme W336F of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 1205 | - |
raffinose | wild type enzyme isoform AgaB, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 2382 | - |
4-nitrophenyl alpha-D-galactopyranoside | wild type enzyme isoform AgaB, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.2.1.22 | 0.05 | - |
raffinose | mutant enzyme W336N of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 0.14 | - |
raffinose | mutant enzyme W336S of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 0.19 | - |
raffinose | mutant enzyme W336A of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 0.7 | - |
4-nitrophenyl alpha-D-galactopyranoside | mutant enzyme W336D of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 3.3 | - |
raffinose | mutant enzyme W336F of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 5.5 | - |
4-nitrophenyl alpha-D-galactopyranoside | mutant enzyme W336S of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 6 | - |
raffinose | wild type enzyme isoform AgaB, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 17.9 | - |
4-nitrophenyl alpha-D-galactopyranoside | mutant enzyme W336A of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 45.5 | - |
4-nitrophenyl alpha-D-galactopyranoside | mutant enzyme W336N of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 64.3 | - |
raffinose | wild type enzyme isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 502 | - |
4-nitrophenyl alpha-D-galactopyranoside | wild type enzyme isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 1562 | - |
4-nitrophenyl alpha-D-galactopyranoside | mutant enzyme W336F of isoform AgaA, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
| 3.2.1.22 | 2870 | - |
4-nitrophenyl alpha-D-galactopyranoside | wild type enzyme isoform AgaB, in 100 mM potassium phosphate buffer, pH 6.5, at 25°C | Geobacillus stearothermophilus | |
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