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Literature summary for 3.2.1.20 extracted from
- Enhancing the thermostability of alpha-glucosidase from Thermoanaerobacter tengcongensis MB4 by single proline substitution (2010), J. Biosci. Bioeng., 110, 12-17.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene gluA, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Caldanaerobacter subterraneus subsp. tengcongensis |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| K285P | site-directed mutagenesis, the mutant shows increased thermostability but no distinct changes in its secondary structure compared to the wild-type enzyme | Caldanaerobacter subterraneus subsp. tengcongensis |
| L152P | site-directed mutagenesis, the mutant shows increased thermostability but no distinct changes in its secondary structure compared to the wild-type enzyme | Caldanaerobacter subterraneus subsp. tengcongensis |
| N208P | site-directed mutagenesis, the mutant shows no increased thermostability compared to the wild-type enzyme | Caldanaerobacter subterraneus subsp. tengcongensis |
| T430P | site-directed mutagenesis, the mutant shows increased thermostability but no distinct changes in its secondary structure compared to the wild-type enzyme | Caldanaerobacter subterraneus subsp. tengcongensis |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.32 | - |
4-nitrophenyl alpha-D-glucopyranoside | pH 5.5, 60°C, mutant K285P | Caldanaerobacter subterraneus subsp. tengcongensis |  |
| 1.08 | - |
4-nitrophenyl alpha-D-glucopyranoside | pH 5.5, 60°C, wild-type enzyme | Caldanaerobacter subterraneus subsp. tengcongensis | |
| 1.61 | - |
4-nitrophenyl alpha-D-glucopyranoside | pH 5.5, 60°C, mutant L152P | Caldanaerobacter subterraneus subsp. tengcongensis | |
| 1.64 | - |
4-nitrophenyl alpha-D-glucopyranoside | pH 5.5, 60°C, mutant T430P | Caldanaerobacter subterraneus subsp. tengcongensis | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Caldanaerobacter subterraneus subsp. tengcongensis | Q8RDL1 | gene gluA | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by affinity chromatography and gel filtration | Caldanaerobacter subterraneus subsp. tengcongensis |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-nitrophenyl alpha-D-glucopyranoside + H2O | - |
Caldanaerobacter subterraneus subsp. tengcongensis | 4-nitrophenol + alpha-D-glucopyranose | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| More | secondary structure of wild-type and mutant enzymes and three-dimensional structure modeling, overview | Caldanaerobacter subterraneus subsp. tengcongensis |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| GluA | - |
Caldanaerobacter subterraneus subsp. tengcongensis |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 60 | - |
assay at | Caldanaerobacter subterraneus subsp. tengcongensis |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 74 | - |
recombinant mutant N208P shows 20% remaining activity, while the wild-type enzyme shows 32% residual activity | Caldanaerobacter subterraneus subsp. tengcongensis |
| 77 | - |
15 min, the recombinant wild-type is almost inactive, while mutants K285P, L152P, and T430P retain 67%, 25%, and 20% residual activity, respectively | Caldanaerobacter subterraneus subsp. tengcongensis |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1.72 | - |
4-nitrophenyl alpha-D-glucopyranoside | pH 5.5, 60°C, mutant K285P | Caldanaerobacter subterraneus subsp. tengcongensis | |
| 4.27 | - |
4-nitrophenyl alpha-D-glucopyranoside | pH 5.5, 60°C, mutant L152P | Caldanaerobacter subterraneus subsp. tengcongensis | |
| 6.64 | - |
4-nitrophenyl alpha-D-glucopyranoside | pH 5.5, 60°C, wild-type enzyme | Caldanaerobacter subterraneus subsp. tengcongensis | |
| 7.25 | - |
4-nitrophenyl alpha-D-glucopyranoside | pH 5.5, 60°C, mutant T430P | Caldanaerobacter subterraneus subsp. tengcongensis | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 5.5 | - |
assay at | Caldanaerobacter subterraneus subsp. tengcongensis |
General Information
| General Information | Comment | Organism |
|---|---|---|
| additional information | Leu152, Lys285, and Thr430 are not only important for the thermostability but also related to the substrate binding and catalytic activity of TtGluA | Caldanaerobacter subterraneus subsp. tengcongensis |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 2.65 | - |
4-nitrophenyl alpha-D-glucopyranoside | pH 5.5, 60°C, mutant L152P | Caldanaerobacter subterraneus subsp. tengcongensis | |
| 4.42 | - |
4-nitrophenyl alpha-D-glucopyranoside | pH 5.5, 60°C, mutant T430P | Caldanaerobacter subterraneus subsp. tengcongensis | |
| 5.37 | - |
4-nitrophenyl alpha-D-glucopyranoside | pH 5.5, 60°C, mutant K285P | Caldanaerobacter subterraneus subsp. tengcongensis | |
| 6.15 | - |
4-nitrophenyl alpha-D-glucopyranoside | pH 5.5, 60°C, wild-type enzyme | Caldanaerobacter subterraneus subsp. tengcongensis | |
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