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Literature summary for 1.1.1.47 extracted from
- Enzymatic reduction of nicotinamide biomimetic cofactors using an engineered glucose dehydrogenase providing a regeneration system for artificial cofactors (2017), ACS Catal., 7, 5202-5208 .
No PubMed abstract available
Application
| Application | Comment | Organism |
|---|---|---|
| biotechnology | enzymatic reduction of the nicotinamide biomimetic cofactors 1-phenethyl-1,4-dihydropyridine-3-carboxamide using glucose dehydrogenase mutant I192T/V306I provides a regeneration system for artificial cofactors. The I192T/V306I mutant enzyme shows 10fold higher activity with 1-phenethyl-1,4-dihydropyridine-3-carboxamide compared with the wild-type enzyme. Using this engineered variant in combination with an enoate reductase from Thermus scotoductus results in an enzyme-coupled regeneration process for biomimetic cofactor without ribonucleotide or ribonucleotide analogue and full conversion of 10 mM 2-methylbut-2-enal with 1-phenethyl-1,4-dihydropyridine-3-carboxamide as cofactor | Saccharolobus solfataricus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| I192T | kcat/Km-values of the mutant enzyme for NAD+ and the biomimetic cofactors are lower than the kcat/Km-values of wild-type enzyme | Saccharolobus solfataricus |
| I192T/V306G | kcat/Km-values of the mutant enzyme for the biomimetic cofactors are higher than the kcat/Km-values of wild-type enzyme. The kcat/Km-value for NAD+ is about 3fold lower than the kcat/Km-value of the wild-type enzyme | Saccharolobus solfataricus |
| I192T/V306I | mutant enzyme shows 10fold higher activity with 1-phenethyl-1,4-dihydropyridine-3-carboxamide compared with the wild-type enzyme. Using this engineered variant in combination with an enoate reductase from Thermus scotoductus results in an enzyme-coupled regeneration process for biomimetic cofactor without ribonucleotide or ribonucleotide analogue and full conversion of 10 mM 2-methylbut-2-enal with 1-phenethyl-1,4-dihydropyridine-3-carboxamide as cofactor | Saccharolobus solfataricus |
| V306G | kcat/Km-values of the mutant enzyme for the biomimetic cofactors are slightly higher than the kcat/Km-values of wild-type enzyme. The kcat/Km-value for NAD+ is about 2fold lower than the kcat/Km-value of the wild-type enzyme | Saccharolobus solfataricus |
| V306I | kcat/Km-values of the mutant enzyme for NAD+ and the biomimetic cofactors are lower than the kcat/Km-values of wild-type enzyme | Saccharolobus solfataricus |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.17 | - |
NAD+ | pH and temperature not specified in the publication, mutant enzyme I192T/V306I | Saccharolobus solfataricus |  |
| 0.36 | - |
NAD+ | pH and temperature not specified in the publication, mutant enzyme I192T | Saccharolobus solfataricus | |
| 0.41 | - |
NAD+ | pH and temperature not specified in the publication, mutant enzyme I192T/V306G | Saccharolobus solfataricus | |
| 0.43 | - |
NAD+ | pH and temperature not specified in the publication, wild-type enzyme | Saccharolobus solfataricus | |
| 0.47 | - |
NAD+ | pH and temperature not specified in the publication, mutant enzyme I192V | Saccharolobus solfataricus | |
| 0.86 | - |
NAD+ | pH and temperature not specified in the publication, mutant enzyme V306I | Saccharolobus solfataricus | |
| 1.67 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, mutant enzyme I192T/V306G | Saccharolobus solfataricus | |
| 2.5 | 3 | 1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192T/V306G | Saccharolobus solfataricus | |
| 2.64 | - |
NAD+ | pH and temperature not specified in the publication, mutant enzyme V306G | Saccharolobus solfataricus | |
| 4.1 | - |
1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192T/V306I | Saccharolobus solfataricus | |
| 4.35 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192T/V306G | Saccharolobus solfataricus | |
| 5.13 | - |
1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme V306I | Saccharolobus solfataricus | |
| 5.49 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, mutant enzyme I192T/V306I | Saccharolobus solfataricus | |
| 7.07 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme V306I | Saccharolobus solfataricus | |
| 8.16 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192T/V306I | Saccharolobus solfataricus | |
| 8.76 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, mutant enzyme V306G | Saccharolobus solfataricus | |
| 10.24 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, wild-type enzyme | Saccharolobus solfataricus | |
| 10.29 | - |
1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme V306G | Saccharolobus solfataricus | |
| 11.32 | - |
1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, wild-type enzyme | Saccharolobus solfataricus | |
| 11.57 | - |
1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192V | Saccharolobus solfataricus | |
| 11.84 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme V306G | Saccharolobus solfataricus | |
| 12.76 | - |
1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192T | Saccharolobus solfataricus | |
| 13.02 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, mutant enzyme I192V | Saccharolobus solfataricus | |
| 13.15 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, mutant enzyme V306I | Saccharolobus solfataricus | |
| 13.87 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192V | Saccharolobus solfataricus | |
| 14.78 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, mutant enzyme I192T | Saccharolobus solfataricus | |
| 15.26 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192T | Saccharolobus solfataricus | |
| 16.33 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, wild-type enzyme | Saccharolobus solfataricus |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Saccharolobus solfataricus | O93715 | - |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| D-glucose + 1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | - |
Saccharolobus solfataricus | D-glucono-1,5-lactone + reduced 1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | - |
? | |
| D-glucose + 1-benzyl-3-carbamoylpyridin-1-ium chloride | - |
Saccharolobus solfataricus | D-glucono-1,5-lactone + reduced 1-benzyl-3-carbamoylpyridin-1-ium chloride | - |
? | |
| D-glucose + 1-phenethyl-1,4-dihydropyridine-3-carboxamide | - |
Saccharolobus solfataricus | D-glucono-1,5-lactone + reduced 1-phenethyl-1,4-dihydropyridine-3-carboxamide | - |
? | |
| D-glucose + NAD+ | - |
Saccharolobus solfataricus | D-glucono-1,5-lactone + NADH + H+ | - |
? | |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.00008 | - |
NAD+ | pH and temperature not specified in the publication, mutant enzyme I192T/V306G | Saccharolobus solfataricus | |
| 0.00023 | - |
1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme V306G | Saccharolobus solfataricus | |
| 0.00047 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme V306G | Saccharolobus solfataricus | |
| 0.00076 | - |
1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192T | Saccharolobus solfataricus | |
| 0.00078 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, mutant enzyme V306G | Saccharolobus solfataricus | |
| 0.00089 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, mutant enzyme I192T | Saccharolobus solfataricus | |
| 0.0009 | - |
1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme V306I | Saccharolobus solfataricus | |
| 0.00095 | - |
NAD+ | pH and temperature not specified in the publication, mutant enzyme I192T/V306I | Saccharolobus solfataricus | |
| 0.00096 | - |
1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, wild-type enzyme | Saccharolobus solfataricus | |
| 0.00126 | - |
1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192V | Saccharolobus solfataricus | |
| 0.00145 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, mutant enzyme V306I | Saccharolobus solfataricus | |
| 0.0016 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, wild-type enzyme | Saccharolobus solfataricus | |
| 0.00162 | - |
NAD+ | pH and temperature not specified in the publication, mutant enzyme V306G | Saccharolobus solfataricus | |
| 0.00174 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, mutant enzyme I192T/V306G | Saccharolobus solfataricus | |
| 0.00246 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, mutant enzyme I192V | Saccharolobus solfataricus | |
| 0.00254 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme 0.00184 I192T/V306G | Saccharolobus solfataricus | |
| 0.00283 | - |
NAD+ | pH and temperature not specified in the publication, mutant enzyme I192T | Saccharolobus solfataricus | |
| 0.00286 | - |
NAD+ | pH and temperature not specified in the publication, mutant enzyme I192V | Saccharolobus solfataricus | |
| 0.00303 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192T | Saccharolobus solfataricus | |
| 0.00396 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, wild-type enzyme | Saccharolobus solfataricus | |
| 0.00398 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192V | Saccharolobus solfataricus | |
| 0.00442 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme V306I | Saccharolobus solfataricus | |
| 0.00678 | - |
NAD+ | pH and temperature not specified in the publication, wild-type enzyme | Saccharolobus solfataricus | |
| 0.00697 | - |
1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192T/V306I | Saccharolobus solfataricus | |
| 0.00838 | - |
NAD+ | pH and temperature not specified in the publication, mutant enzyme V306I | Saccharolobus solfataricus | |
| 0.009 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, mutant enzyme I192T/V306I | Saccharolobus solfataricus | |
| 0.042 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192T/V306I | Saccharolobus solfataricus |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.00002 | - |
1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme V306G | Saccharolobus solfataricus | |
| 0.00004 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme V306G | Saccharolobus solfataricus | |
| 0.00006 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, mutant enzyme I192T | Saccharolobus solfataricus | |
| 0.00006 | - |
1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192T | Saccharolobus solfataricus | |
| 0.00009 | - |
1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, wild-type enzyme | Saccharolobus solfataricus | |
| 0.00009 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, mutant enzyme V306G | Saccharolobus solfataricus | |
| 0.00011 | - |
1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192V | Saccharolobus solfataricus | |
| 0.00011 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, mutant enzyme V306I | Saccharolobus solfataricus | |
| 0.00016 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, wild-type enzyme | Saccharolobus solfataricus | |
| 0.00018 | - |
1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme V306I | Saccharolobus solfataricus | |
| 0.00019 | - |
NAD+ | pH and temperature not specified in the publication, mutant enzyme I192T/V306G | Saccharolobus solfataricus | |
| 0.00019 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, mutant enzyme I192V | Saccharolobus solfataricus | |
| 0.0002 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192T | Saccharolobus solfataricus | |
| 0.00024 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, wild-type enzyme | Saccharolobus solfataricus | |
| 0.00029 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192V | Saccharolobus solfataricus | |
| 0.00058 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192T/V306G | Saccharolobus solfataricus | |
| 0.00061 | - |
NAD+ | pH and temperature not specified in the publication, mutant enzyme V306G | Saccharolobus solfataricus | |
| 0.00063 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme V306I | Saccharolobus solfataricus | |
| 0.00073 | - |
1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192T/V306G | Saccharolobus solfataricus | |
| 0.000974 | - |
NAD+ | pH and temperature not specified in the publication, mutant enzyme V306I | Saccharolobus solfataricus | |
| 0.00104 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, mutant enzyme I192T/V306G | Saccharolobus solfataricus | |
| 0.00164 | - |
1-benzyl-3-carbamoylpyridin-1-ium chloride | pH and temperature not specified in the publication, mutant enzyme I192T/V306I | Saccharolobus solfataricus | |
| 0.0017 | - |
1-(3-phenylpropyl)-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192T/V306I | Saccharolobus solfataricus | |
| 0.00517 | - |
1-phenethyl-1,4-dihydropyridine-3-carboxamide | pH and temperature not specified in the publication, mutant enzyme I192T/V306I | Saccharolobus solfataricus | |
| 0.00565 | - |
NAD+ | pH and temperature not specified in the publication, mutant enzyme I192T/V306I | Saccharolobus solfataricus | |
| 0.00613 | - |
NAD+ | pH and temperature not specified in the publication, mutant enzyme I192V | Saccharolobus solfataricus | |
| 0.00778 | - |
NAD+ | pH and temperature not specified in the publication, mutant enzyme I192T | Saccharolobus solfataricus | |
| 0.01594 | - |
NAD+ | pH and temperature not specified in the publication, wild-type enzyme | Saccharolobus solfataricus | |
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