Cloned (Comment) | Organism |
---|---|
overexpression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli | Synechocystis sp. |
Protein Variants | Comment | Organism |
---|---|---|
W204A | site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to the wild-type enzyme | Synechocystis sp. |
W204F | site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to the wild-type enzyme, the mutant is active with the substrate analogue 1,2-dideoxy-D-threo-3-hexulose 6-phosphate in contrast to the wild-type enzyme | Synechocystis sp. |
W204L | site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to the wild-type enzyme | Synechocystis sp. |
W204V | site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to the wild-type enzyme | Synechocystis sp. |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
1,2-dideoxy-D-threo-3-hexulose 6-phosphate | weak, competitive inhibition, a highly conserved tryptophan residue in the flexible loop is blocked by the substrate analog | Synechocystis sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.17 | - |
1-deoxy-D-xylulose 5-phosphate | pH 7.8, 37°C, recombinant wild-type enzyme | Synechocystis sp. | |
0.42 | - |
1-deoxy-D-xylulose 5-phosphate | pH 7.8, 37°C, recombinant mutant W204F | Synechocystis sp. | |
1.3 | - |
1,2-dideoxy-D-threo-3-hexulose 6-phosphate | pH 7.8, 37°C, recombinant mutant W204F | Synechocystis sp. | |
2.9 | - |
1-deoxy-D-xylulose 5-phosphate | pH 7.8, 37°C, recombinant mutant W204L | Synechocystis sp. | |
5 | - |
1-deoxy-D-xylulose 5-phosphate | pH 7.8, 37°C, recombinant mutant W204V | Synechocystis sp. | |
12 | - |
1-deoxy-D-xylulose 5-phosphate | pH 7.8, 37°C, recombinant mutant W204A | Synechocystis sp. |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | required | Synechocystis sp. |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
1-deoxy-D-xylulose 5-phosphate + NADPH + H+ | Synechocystis sp. | - |
2-C-methyl-D-erythritol 4-phosphate + NADP+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Synechocystis sp. | - |
strain PCC6803 | - |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli by metal ion affinity chromatography | Synechocystis sp. |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
2-C-methyl-D-erythritol 4-phosphate + NADP+ = 1-deoxy-D-xylulose 5-phosphate + NADPH + H+ | the conserved active site residue Trp204 is important in catalysis | Synechocystis sp. |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Synechocystis sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1,2-dideoxy-D-threo-3-hexulose 6-phosphate + NADPH + H+ | i.e. 1-methyl-1-deoxy-D-xylulose 5-phosphate or M-DXP, a substrate analogue whichis inactive with the wild-type but active with mutant W204F | Synechocystis sp. | 2-C-ethyl-D-erythritol 4-phosphate + NADP+ | - |
? | |
1-deoxy-D-xylulose 5-phosphate + NADPH + H+ | - |
Synechocystis sp. | 2-C-methyl-D-erythritol 4-phosphate + NADP+ | - |
? | |
1-deoxy-D-xylulose 5-phosphate + NADPH + H+ | i.e. DXP | Synechocystis sp. | 2-C-methyl-D-erythritol 4-phosphate + NADP+ | - |
? | |
additional information | substrate synthesis and product identification method, overview | Synechocystis sp. | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
DXP reductoisomerase | - |
Synechocystis sp. |
DXR | - |
Synechocystis sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Synechocystis sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.04 | - |
1,2-dideoxy-D-threo-3-hexulose 6-phosphate | pH 7.8, 37°C, recombinant mutant W204F | Synechocystis sp. | |
0.16 | - |
1-deoxy-D-xylulose 5-phosphate | pH 7.8, 37°C, recombinant mutant W204A | Synechocystis sp. | |
0.26 | - |
1-deoxy-D-xylulose 5-phosphate | pH 7.8, 37°C, recombinant mutant W204L | Synechocystis sp. | |
0.54 | - |
1-deoxy-D-xylulose 5-phosphate | pH 7.8, 37°C, recombinant mutant W204V | Synechocystis sp. | |
10 | - |
1-deoxy-D-xylulose 5-phosphate | pH 7.8, 37°C, recombinant mutant W204F | Synechocystis sp. | |
17 | - |
1-deoxy-D-xylulose 5-phosphate | pH 7.8, 37°C, recombinant wild-type enzyme | Synechocystis sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.8 | - |
assay at | Synechocystis sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | - |
Synechocystis sp. |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.63 | - |
1,2-dideoxy-D-threo-3-hexulose 6-phosphate | pH 7.8, 37°C, recombinant wild-type enzyme | Synechocystis sp. |