Cloned (Comment) | Organism |
---|---|
amplified gene cloned into the pGEM-T Easy vector, heterologous expression with a His-tag alternatively placed in the N- or C-terminus in Escherichia coli BL21 Star (DE3) cells | Malus domestica |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
(NH4)2SO4 | - |
Malus domestica | |
KCl | - |
Malus domestica | |
additional information | no significant differences in activity when ATP, ADP, AMP, ADP-glucose, UTP, UDP-glucose, inorganic phosphate or diphosphate (5 mM each), 10% (v/v) glycerol, 500 mM glucitol or 20% (w/v) sucrose are added to the enzyme assay mixture | Malus domestica | |
NaCl | 200 mM cause about 65% inhibition of the enzyme activity | Malus domestica | |
NH4Cl | - |
Malus domestica |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
soluble | level of soluble recombinant enzyme is much higher in the N-term tagged protein than in the C-term tagged one. In the C-term tagged one the majority of the protein is in the insoluble form | Malus domestica | - |
- |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
34900 | - |
sequence analysis, A6PRase | Malus domestica |
39000 | - |
gel filtration, C-term tagged enzyme | Malus domestica |
68000 | - |
gel filtration, N-term tagged enzyme | Malus domestica |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Malus domestica | P28475 | - |
- |
Purification (Comment) | Organism |
---|---|
one-step protein purification by immobilized metal affinity chromatography, N-term tagged protein 95% pure, C-term tagged one 75% pure | Malus domestica |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
leaf | - |
Malus domestica | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.017 | - |
C-term tagged enzyme, purified enzyme | Malus domestica |
0.136 | - |
N-term tagged enzyme, crude extract | Malus domestica |
1.25 | - |
N-term tagged enzyme, 9.2fold purified | Malus domestica |
Storage Stability | Organism |
---|---|
-80°C, 25 mM Tris-HCl buffer, pH 8.0, 300 mM NaCl, 1 mM beta-mercaptoethanol, 10% (v/v) glycerol, 1 mM EDTA, 0.5 mM DTT, 6 months | Malus domestica |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-glucitol-6-phosphate | - |
Malus domestica | D-glucose 6-phosphate + NADPH + H+ | - |
r | |
D-glucose 6-phosphate + NADPH + H+ | - |
Malus domestica | D-glucitol-6-phosphate + NADP+ | - |
r | |
additional information | both, the N- and the C-term tagged enzymes exhibit similar affinity toward substrates | Malus domestica | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | gel filtration, N-term tagged enzyme | Malus domestica |
monomer | gel filtration, C-term tagged enzyme | Malus domestica |
Synonyms | Comment | Organism |
---|---|---|
A6PRase | - |
Malus domestica |
aldose-6-phosphate reductase | - |
Malus domestica |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0011 | - |
D-glucitol-6-phosphate | C-term tagged enzyme | Malus domestica | |
0.009 | - |
D-glucose 6-phosphate | C-term tagged enzyme | Malus domestica | |
0.222 | - |
D-glucitol-6-phosphate | N-term tagged enzyme | Malus domestica | |
0.7 | - |
D-glucose 6-phosphate | N-term tagged enzyme | Malus domestica |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 10 | N- and the C-term tagged enzymes. Both recombinant enzymes similarly show optimal activity in a broad pH range in the direction of D-glucose 6-phosphate reduction, and a narrow range (pH 9.0-10.0) in the opposite oxidative direction | Malus domestica |