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Literature summary extracted from
- High-throughput screening of coenzyme preference change of thermophilic 6-phosphogluconate dehydrogenase from NADP+ to NAD+ (2016), Sci. Rep., 6, 32644 .
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.1.1.44 | gene 6pgdh, recombinant expression in Escherichia coli TOP10, recombinant overexpression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Moorella thermoacetica |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 1.1.1.44 | A30C/R31I/T32K | saturation mutagenesis, the mutant shows better kinetics and increased activity for NAD+ compared to the wild-type enzyme | Moorella thermoacetica |
| 1.1.1.44 | A30D/R31I/T32I | saturation mutagenesis, the mutant shows much better kinetics and strongly increased activity for NAD+ compared to the wild-type enzyme | Moorella thermoacetica |
| 1.1.1.44 | A30E/R31I/T32D | saturation mutagenesis, the mutant shows better kinetics and increased activity for NAD+ compared to the wild-type enzyme | Moorella thermoacetica |
| 1.1.1.44 | additional information | high-throughput screening of the two-site saturation mutagenesis library A30/T32 in Escherichia coli TOP10 by using a pair of degenerate primers 30_32_NNK_F/30_32_NNK_R, enzyme mutant identification and selection by heat treatment, usage of the dual T7-tac promoter and a colorimetric assay, method optimization, overview | Moorella thermoacetica |
| 1.1.1.44 | R31I | saturation mutagenesis, the mutant shows slightly better kinetics and slightly increased activity for NAD+ compared to the wild-type enzyme | Moorella thermoacetica |
| 1.1.1.44 | R31I/T32G | saturation mutagenesis, the mutant shows slightly better kinetics and slightly increased activity for NAD+ compared to the wild-type enzyme | Moorella thermoacetica |
| 1.1.1.44 | R31T | saturation mutagenesis, the mutant shows slightly better kinetics and slightly increased activity for NAD+ compared to the wild-type enzyme | Moorella thermoacetica |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.1.1.44 | 0.0119 | - |
NAD+ | recombinant His-tagged mutant A30D/R31I/T32I, pH 7.5, 50°C | Moorella thermoacetica |  |
| 1.1.1.44 | 0.0139 | - |
NADP+ | recombinant His-tagged wild-type enzyme, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 0.0265 | - |
NADP+ | recombinant His-tagged mutant R31I, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 0.0355 | - |
NADP+ | recombinant His-tagged mutant R31T, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 0.1044 | - |
NADP+ | recombinant His-tagged mutant R31I/T32G, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 0.127 | - |
NAD+ | recombinant His-tagged mutant A30E/R31I/T32D, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 0.228 | - |
NADP+ | recombinant His-tagged mutant A30D/R31I/T32I, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 0.354 | - |
NAD+ | recombinant His-tagged mutant R31I, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 0.362 | - |
NAD+ | recombinant His-tagged mutant R31I/T32G, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 0.404 | - |
NAD+ | recombinant His-tagged mutant A30C/R31I/T32K, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 0.605 | - |
NAD+ | recombinant His-tagged mutant R31T, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 0.66 | - |
NADP+ | recombinant His-tagged mutant A30E/R31I/T32D, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 0.698 | - |
NADP+ | recombinant His-tagged mutant A30C/R31I/T32K, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 1.397 | - |
NAD+ | recombinant His-tagged wild-type enzyme, pH 7.5, 50°C | Moorella thermoacetica | |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.1.1.44 | Mg2+ | required | Moorella thermoacetica | |
| 1.1.1.44 | Mn2+ | required | Moorella thermoacetica | |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.1.1.44 | 6-phospho-D-gluconate + NADP+ | Moorella thermoacetica | - |
D-ribulose 5-phosphate + CO2 + NADPH + H+ | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.1.1.44 | Moorella thermoacetica | Q2RIZ2 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.1.1.44 | recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Moorella thermoacetica |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.1.1.44 | 6-phospho-D-gluconate + NAD+ | very low activity with NAD+ with the wild-type enzyme, low activity with mutants R31T, R31I, and R31I/T32G, higher activity with mutants A30C/R31I/T32K and A30E/R31I/T32D, and high activity with mutant A30D/R31I/T32I | Moorella thermoacetica | D-ribulose 5-phosphate + CO2 + NADH + H+ | - |
? | |
| 1.1.1.44 | 6-phospho-D-gluconate + NADP+ | - |
Moorella thermoacetica | D-ribulose 5-phosphate + CO2 + NADPH + H+ | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.1.1.44 | More | three-dimensional structure homology modeling of wild-type 6PGDH from Moorella thermoacetica and mutants based on the structure of human enzyme 6PGDH, PDB ID 2JKV | Moorella thermoacetica |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.1.1.44 | 6-phosphogluconate dehydrogenase | - |
Moorella thermoacetica |
| 1.1.1.44 | 6PGDH | - |
Moorella thermoacetica |
| 1.1.1.44 | Moth_1283 | - |
Moorella thermoacetica |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.1.1.44 | 50 | - |
assay at | Moorella thermoacetica |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.1.1.44 | 1.81 | - |
NADP+ | recombinant His-tagged mutant A30D/R31I/T32I, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 3.1 | - |
NADP+ | recombinant His-tagged mutant A30E/R31I/T32D, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 5.75 | - |
NAD+ | recombinant His-tagged mutant A30D/R31I/T32I, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 6 | - |
NAD+ | recombinant His-tagged mutant A30C/R31I/T32K, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 6.23 | - |
NADP+ | recombinant His-tagged mutant A30C/R31I/T32K, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 8.73 | - |
NADP+ | recombinant His-tagged wild-type enzyme, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 9.12 | - |
NAD+ | recombinant His-tagged mutant R31T, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 10.8 | - |
NAD+ | recombinant His-tagged mutant A30E/R31I/T32D, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 10.83 | - |
NADP+ | recombinant His-tagged mutant R31T, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 11.53 | - |
NADP+ | recombinant His-tagged mutant R31I, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 11.9 | - |
NADP+ | recombinant His-tagged mutant R31I/T32G, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 12.6 | - |
NAD+ | recombinant His-tagged wild-type enzyme, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 12.94 | - |
NAD+ | recombinant His-tagged mutant R31I/T32G, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 15.01 | - |
NAD+ | recombinant His-tagged mutant R31I, pH 7.5, 50°C | Moorella thermoacetica | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.1.1.44 | 7.5 | - |
assay at | Moorella thermoacetica |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.1.1.44 | NAD+ | active with enzyme mutants, not with the wild-type enzyme | Moorella thermoacetica | |
| 1.1.1.44 | NADP+ | specific for, natural coenzyme is NADP+ | Moorella thermoacetica | |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 1.1.1.44 | additional information | three-dimensional structure homology modeling of wild-type 6PGDH from Moorella thermoacetica and mutants based on the structure of human enzyme 6PGDH, PDB ID 2JKV | Moorella thermoacetica |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.1.1.44 | 4.7 | - |
NADP+ | recombinant His-tagged mutant A30E/R31I/T32D, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 7.9 | - |
NADP+ | recombinant His-tagged mutant A30D/R31I/T32I, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 8.9 | - |
NADP+ | recombinant His-tagged mutant A30C/R31I/T32K, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 9 | - |
NAD+ | recombinant His-tagged wild-type enzyme, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 14.8 | - |
NAD+ | recombinant His-tagged mutant A30C/R31I/T32K, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 15 | - |
NAD+ | recombinant His-tagged mutant R31T, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 35.8 | - |
NAD+ | recombinant His-tagged mutant R31I/T32G, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 42.4 | - |
NAD+ | recombinant His-tagged mutant R31I, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 85.1 | - |
NAD+ | recombinant His-tagged mutant A30E/R31I/T32D, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 114 | - |
NADP+ | recombinant His-tagged mutant R31I/T32G, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 305.6 | - |
NADP+ | recombinant His-tagged mutant R31T, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 435 | - |
NADP+ | recombinant His-tagged mutant R31I, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 484.2 | - |
NAD+ | recombinant His-tagged mutant A30D/R31I/T32I, pH 7.5, 50°C | Moorella thermoacetica | |
| 1.1.1.44 | 628.8 | - |
NADP+ | recombinant His-tagged wild-type enzyme, pH 7.5, 50°C | Moorella thermoacetica | |
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