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Literature summary for 1.1.1.360 extracted from
- Structural insight into glucose dehydrogenase from the thermoacidophilic archaeon Thermoplasma volcanium (2014), Acta Crystallogr. Sect. D, 70, 1271-1280.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli BL21(DE3) cells | Thermoplasma volcanium |
| expression in Escherichia coli | Thermoplasma volcanium |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| crystal structures of substrate/cofactor-free tvGlcDH and of a tvGlcDH T277F mutant in a binary complex with NADP and in a ternary complex with D-glucose and nicotinic acid adenine dinucleotide phosphate, an NADP analogue, are determined at resolutions of 2.6, 2.25 and 2.33 A | Thermoplasma volcanium |
| in a binary complex with NADP+ and in a ternary complex with D-glucose and nicotinic acid adenine dinucleotide phosphate, sitting drop vapor diffusion method, using 30% (w/v) 1,2-propanediol, 20% (w/v) polyethylene glycol 400 (PEG 400), 0.1 M HEPES pH 7.5 | Thermoplasma volcanium |
| ultrafiltration, sitting-drop vapour-diffusion method, crystal structures of substrate/cofactor-free enzyme and of a T277F mutant enzyme in a binary complex with NADP+ and in a ternary complex with D-glucose and nicotinic acid adenine dinucleotide phosphate, an NADP+ analogue, are determined at resolutions of 2.6, 2.25 and 2.33 A, respectively | Thermoplasma volcanium |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D306N | enhancement of the activity towards D-xylose is not observed, mutation reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| D306N | improvement of the reactivity towards D-xylose can not be achieved, while the mutations reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| E156Q | enhancement of the activity towards D-xylose is not observed, mutation reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| E156Q | improvement of the reactivity towards D-xylose can not be achieved, while the mutations reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| E296H | enhancement of the activity towards D-xylose is not observed, mutation reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| E296H | improvement of the reactivity towards D-xylose can not be achieved, while the mutations reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| K159A | enhancement of the activity towards D-xylose is not observed, mutation reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| K159A | improvement of the reactivity towards D-xylose can not be achieved, while the mutations reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| K159A/N160D | enhancement of the activity towards D-xylose is not observed, mutation reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| K159A/N160D | improvement of the reactivity towards D-xylose can not be achieved, while the mutations reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| N160D | enhancement of the activity towards D-xylose is not observed, mutation reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| N160D | improvement of the reactivity towards D-xylose can not be achieved, while the mutations reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| T277F | crystal structure of the T277F mutant enzyme in a binary complex with NADP+ and in a ternary complex with D-glucose and nicotinic acid adenine dinucleotide phosphate. The Vmax for the T277F mutant is about 12% of that for the wild-type enzyme | Thermoplasma volcanium |
| T277F | crystallization of the T277F mutant enzyme in a binary complex with NADP and in a ternary complex with D-glucose and nicotinic acid adenine dinucleotide phosphate, mutation reduces the D-glucose oxidation activity, the Vmax for the T277F mutant is about 12% of that for the wild-type enzyme | Thermoplasma volcanium |
| V93N | enhancement of the activity towards D-xylose is not observed, mutation reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| V93N | the Vmax value for D-glucose oxidation by the purified V93N mutant is about 71% of the Vmax value for the wild-type enzyme. Enhancement of the activity towards D-xylose is not observed | Thermoplasma volcanium |
| V93N/D306N | enhancement of the activity towards D-xylose is not observed, mutation reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| V93N/D306N | improvement of the reactivity towards D-xylose can not be achieved, while the mutations reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| V93N/E156Q/K159A/N160D/D306N/E296H | activity is completely abolished | Thermoplasma volcanium |
| V93N/E156Q/K159A/N160D/D306N/E296H | mutation completely abolished the activity of the enzyme | Thermoplasma volcanium |
| V93N/K159A | enhancement of the activity towards D-xylose is not observed, mutation reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| V93N/K159A | improvement of the reactivity towards D-xylose can not be achieved, while the mutations reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| V93N/K159A/N160D | enhancement of the activity towards D-xylose is not observed, mutation reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| V93N/K159A/N160D | improvement of the reactivity towards D-xylose can not be achieved, while the mutations reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| V93N/N160D | enhancement of the activity towards D-xylose is not observed, mutation reduces the D-glucose oxidation activity | Thermoplasma volcanium |
| V93N/N160D | improvement of the reactivity towards D-xylose can not be achieved, while the mutations reduces the D-glucose oxidation activity | Thermoplasma volcanium |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.14 | - |
NADP+ | pH 6.5, 55°C, wild-type enzyme | Thermoplasma volcanium |  |
| 0.14 | - |
NADP+ | pH 6.5, 55°C, wild-type enzyme or mutant enzyme T277F | Thermoplasma volcanium | |
| 0.14 | - |
NADP+ | with D-glucose as cosubstrate, at pH 6.5 and 35°C | Thermoplasma volcanium | |
| 7.1 | - |
D-glucopyranose | pH 6.5, 55°C, wild-type enzyme | Thermoplasma volcanium | |
| 7.1 | - |
D-glucose | with NADP+ as cosubstrate, at pH 6.5 and 35°C | Thermoplasma volcanium | |
| 8.7 | - |
D-galactopyranose | pH 6.5, 55°C, wild-type enzyme | Thermoplasma volcanium | |
| 8.7 | - |
D-galactose | with NADP+ as cosubstrate, at pH 6.5 and 35°C | Thermoplasma volcanium | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 41311 | - |
4 * 41311, calculated from sequence | Thermoplasma volcanium |
| 42000 | - |
4 * 42000, SDS-PAGE | Thermoplasma volcanium |
| 158000 | - |
gel filtration | Thermoplasma volcanium |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| D-galactose + NADP+ | Thermoplasma volcanium | the activity towards D-galactose is 58% of that observed towards D-glucose | ? | - |
? | |
| D-galactose + NADP+ | Thermoplasma volcanium DSM 4299 | the activity towards D-galactose is 58% of that observed towards D-glucose | ? | - |
? | |
| D-glucose + NADP+ | Thermoplasma volcanium | - |
D-gluconate + NADPH + H+ | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Thermoplasma volcanium | Q979W2 | - |
- |
| Thermoplasma volcanium DSM 4299 | Q979W2 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Thermoplasma volcanium |
| nickel-charged chelating Sepharose column chromatography and Superdex 200 gel filtration | Thermoplasma volcanium |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 58 | - |
pH 6.5, 55°C, D-glucose oxidation, T277F mutant | Thermoplasma volcanium |
| 356 | - |
pH 6.5, 55°C, D-glucose oxidation, V93N mutant | Thermoplasma volcanium |
| 356 | - |
pH 6.5, 55°C, mutant enzyme V93N | Thermoplasma volcanium |
| 498 | - |
pH 6.5, 55°C, D-glucose oxidation, wild-type enzyme | Thermoplasma volcanium |
| 498 | - |
pH 6.5, 55°C, wild-type enzyme | Thermoplasma volcanium |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| D-galactopyranose + NADP+ | with NADP+ (2 mM) as the electron acceptor, the enzyme acts exclusively on D-glucose and D-galactose (100 mM substrate), and the activity towards D-galactose is 58% of that observed towards D-glucose. The activities with D-xylose, L-arabinose, D-ribose, D-fructose and D-mannose are all less than 4% of that with D-glucose | Thermoplasma volcanium | D-galactono-1,5-lactone + NADPH + H+ | - |
? | |
| D-galactopyranose + NADP+ | with NADP+ as the electron acceptor, the enzyme acts exclusively on D-glucose and D-galactose, and the activity towards D-galactose is 58% of that observed towards D-glucose. The activities with D-xylose, L-arabinose, D-ribose, D-fructose and D-mannose are all less than 4% of that with D-glucose | Thermoplasma volcanium | D-galactono-1,5-lactone + NADPH + H+ | - |
? | |
| D-galactopyranose + NADP+ | with NADP+ (2 mM) as the electron acceptor, the enzyme acts exclusively on D-glucose and D-galactose (100 mM substrate), and the activity towards D-galactose is 58% of that observed towards D-glucose. The activities with D-xylose, L-arabinose, D-ribose, D-fructose and D-mannose are all less than 4% of that with D-glucose | Thermoplasma volcanium DSM 4299 | D-galactono-1,5-lactone + NADPH + H+ | - |
? | |
| D-galactopyranose + NADP+ | with NADP+ as the electron acceptor, the enzyme acts exclusively on D-glucose and D-galactose, and the activity towards D-galactose is 58% of that observed towards D-glucose. The activities with D-xylose, L-arabinose, D-ribose, D-fructose and D-mannose are all less than 4% of that with D-glucose | Thermoplasma volcanium DSM 4299 | D-galactono-1,5-lactone + NADPH + H+ | - |
? | |
| D-galactose + NADP+ | the activity towards D-galactose is 58% of that observed towards D-glucose | Thermoplasma volcanium | ? | - |
? | |
| D-galactose + NADP+ | the activity towards D-galactose is 58% of that observed towards D-glucose | Thermoplasma volcanium DSM 4299 | ? | - |
? | |
| D-glucopyranose + NADP+ | with NADP+ (2 mM) as the electron acceptor, the enzyme acts exclusively on D-glucose and D-galactose (100 mM substrate), and the activity towards D-galactose is 58% of that observed towards D-glucose. The activities with D-xylose, L-arabinose, D-ribose, D-fructose and D-mannose are all less than 4% of that with D-glucose | Thermoplasma volcanium | D-glucono-1,5-lactone + NADPH + H+ | - |
? | |
| D-glucopyranose + NADP+ | with NADP+ as the electron acceptor, the enzyme acts exclusively on D-glucose and D-galactose, and the activity towards D-galactose is 58% of that observed towards D-glucose. The activities with D-xylose, L-arabinose, D-ribose, D-fructose and D-mannose are all less than 4% of that with D-glucose | Thermoplasma volcanium | D-glucono-1,5-lactone + NADPH + H+ | - |
? | |
| D-glucose + NADP+ | - |
Thermoplasma volcanium | D-gluconate + NADPH + H+ | - |
? | |
| additional information | the enzyme does not utilize aldopentoses such as D-xylose | Thermoplasma volcanium | ? | - |
? | |
| additional information | the enzyme does not utilize D-xylose , L-arabinose, D-ribose, D-fructose, and D-mannose as substrates | Thermoplasma volcanium | ? | - |
? | |
| additional information | the enzyme does not utilize aldopentoses such as D-xylose | Thermoplasma volcanium DSM 4299 | ? | - |
? | |
| additional information | the enzyme does not utilize D-xylose , L-arabinose, D-ribose, D-fructose, and D-mannose as substrates | Thermoplasma volcanium DSM 4299 | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homotetramer | 4 * 42000, SDS-PAGE | Thermoplasma volcanium |
| homotetramer | 4 * 41311, calculated from sequence | Thermoplasma volcanium |
| homotetramer | 4 * 41311, calculated from amino acid sequence | Thermoplasma volcanium |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| GlcDH | - |
Thermoplasma volcanium |
| TvGlcD | - |
Thermoplasma volcanium |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 55 | - |
assay at | Thermoplasma volcanium |
| 70 | - |
- |
Thermoplasma volcanium |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | - |
the enzyme retains more than 90% of its activity after incubation for 10 min at temperatures of up to 40°C | Thermoplasma volcanium |
| 60 | - |
10 min, retains more than 90% of its activity | Thermoplasma volcanium |
| 60 | - |
10 min, the enzyme retains more than 90% of its activity | Thermoplasma volcanium |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6.5 | - |
- |
Thermoplasma volcanium |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 9 | 60°C, 10 min, stable | Thermoplasma volcanium |
| 6 | 9 | 10 min, at 60°C, no loss of activity | Thermoplasma volcanium |
| 6 | 9 | the enzyme loses no activity when incubated at pH values between 6.0 and 9.0 for 10 min at 40°C | Thermoplasma volcanium |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| NAD+ | the enzyme utilizes NAD+ as a cofactor with nearly the same substrate specificity as NADP, but the electron-acceptor activity of NAD+ (2 mM) is only 4% of that observed with NADP+ (at 100 mM D-glucose) | Thermoplasma volcanium | |
| NADP+ | the electron-acceptor activity of NAD+ (2 mM) is only 4% of that observed with NADP+ (at 100 mM D-glucose) | Thermoplasma volcanium | |
| NADP+ | the enzyme utilizes NAD+ as a cofactor with nearly the same substrate specificity as NADP, but the electron-acceptor activity of NAD+ (2 mM) is only 4% of that observed with NADP+ (at 100 mM D-glucose) | Thermoplasma volcanium | |
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