Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli ER2566 cells | Thermus thermophilus |
Protein Variants | Comment | Organism |
---|---|---|
D138A | the mutant shows 59% activity compared to the wild type enzyme | Thermus thermophilus |
E132A | the mutant shows 1.9% activity compared to the wild type enzyme | Thermus thermophilus |
E132D | the mutant shows 0.4% activity compared to the wild type enzyme | Thermus thermophilus |
E132K | inactive | Thermus thermophilus |
E132Q | inactive | Thermus thermophilus |
E132W | inactive | Thermus thermophilus |
E67A | the mutant shows 2.5% activity compared to the wild type enzyme | Thermus thermophilus |
E67D | the mutant shows 3.7% activity compared to the wild type enzyme | Thermus thermophilus |
E67K | inactive | Thermus thermophilus |
E67Q | the mutant shows 0.8% activity compared to the wild type enzyme | Thermus thermophilus |
E67W | inactive | Thermus thermophilus |
H122A | the mutant shows 2.8% activity compared to the wild type enzyme | Thermus thermophilus |
H122D | inactive | Thermus thermophilus |
H122K | inactive | Thermus thermophilus |
H122Q | the mutant shows 2.0% activity compared to the wild type enzyme | Thermus thermophilus |
H122W | inactive | Thermus thermophilus |
H50A | the mutant shows 2.2% activity compared to the wild type enzyme | Thermus thermophilus |
H50D | inactive | Thermus thermophilus |
H50K | inactive | Thermus thermophilus |
H50Q | the mutant shows 2.0% activity compared to the wild type enzyme | Thermus thermophilus |
H50W | inactive | Thermus thermophilus |
K37A | the mutant shows 21% activity compared to the wild type enzyme | Thermus thermophilus |
K65A | the mutant shows 15% activity compared to the wild type enzyme | Thermus thermophilus |
L18A | the mutant shows 78% activity compared to the wild type enzyme | Thermus thermophilus |
L39A | the mutant shows 84% activity compared to the wild type enzyme | Thermus thermophilus |
Q48A | the mutant shows 2.0% activity compared to the wild type enzyme | Thermus thermophilus |
Q48D | inactive | Thermus thermophilus |
Q48K | inactive | Thermus thermophilus |
Q48N | inactive | Thermus thermophilus |
Q48W | inactive | Thermus thermophilus |
R11A | the mutant shows 260% activity compared to the wild type enzyme | Thermus thermophilus |
R142A | the mutant shows 2.6% activity compared to the wild type enzyme | Thermus thermophilus |
R142D | inactive | Thermus thermophilus |
R142K | inactive | Thermus thermophilus |
R142N | inactive | Thermus thermophilus |
R142W | inactive | Thermus thermophilus |
W13A | the mutant shows 0.4% activity compared to the wild type enzyme | Thermus thermophilus |
W13D | inactive | Thermus thermophilus |
W13F | the mutant shows 41% activity compared to the wild type enzyme | Thermus thermophilus |
W13H | the mutant shows 52% activity compared to the wild type enzyme | Thermus thermophilus |
W13K | inactive | Thermus thermophilus |
W13Q | inactive | Thermus thermophilus |
W13Y | the mutant shows 68% activity compared to the wild type enzyme | Thermus thermophilus |
W69A | the mutant shows 50% activity compared to the wild type enzyme | Thermus thermophilus |
Y124A | the mutant shows 52% activity compared to the wild type enzyme | Thermus thermophilus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ba2+ | about 30% inhibition at 0.5 mM | Thermus thermophilus | |
EDTA | complete inhibition at 0.5 mM | Thermus thermophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.18 | - |
D-mannose 6-phosphate | mutant enzyme E132A, pH 7.0, 80°C | Thermus thermophilus | |
0.18 | - |
D-mannose 6-phosphate | mutant enzyme E67A, pH 7.0, 80°C | Thermus thermophilus | |
0.21 | - |
D-mannose 6-phosphate | wild type enzyme, pH 7.0, 80°C | Thermus thermophilus | |
0.22 | - |
D-fructose 6-phosphate | wild type enzyme, pH 7.0, 80°C | Thermus thermophilus | |
0.27 | - |
D-mannose 6-phosphate | mutant enzyme H122A, pH 7.0, 80°C | Thermus thermophilus | |
0.33 | - |
D-mannose 6-phosphate | mutant enzyme H50A, pH 7.0, 80°C | Thermus thermophilus | |
0.53 | - |
D-mannose 6-phosphate | mutant enzyme Q48A, pH 7.0, 80°C | Thermus thermophilus | |
0.85 | - |
D-mannose 6-phosphate | mutant enzyme W13A, pH 7.0, 80°C | Thermus thermophilus | |
1.29 | - |
D-mannose 6-phosphate | mutant enzyme R142A, pH 7.0, 80°C | Thermus thermophilus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | the enzyme is not stimulated by Ca2+ | Thermus thermophilus | |
Co2+ | about 230% activity at 0.5 mM | Thermus thermophilus | |
Cu2+ | about 135% activity at 0.5 mM | Thermus thermophilus | |
Fe2+ | about 125% activity at 0.5 mM | Thermus thermophilus | |
Mg2+ | about 175% activity at 0.5 mM | Thermus thermophilus | |
Mn2+ | about 175% activity at 0.5 mM | Thermus thermophilus | |
Zn2+ | Zn2+ is present at one molecule per monomer, the enzyme has about 240% activity in the presence of 0.5 mM Zn2+ | Thermus thermophilus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
29000 | - |
1 * 29000, SDS-PAGE | Thermus thermophilus |
29054 | - |
1 * 29054, calculated from amino acid sequence | Thermus thermophilus |
29100 | - |
gel filtration | Thermus thermophilus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermus thermophilus | Q5SIM4 | - |
- |
Thermus thermophilus KCCM 40879 | Q5SIM4 | - |
- |
Purification (Comment) | Organism |
---|---|
HisTrap HP column chromatography | Thermus thermophilus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.01 | - |
wild type enzyme, using ribose 5-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
0.3 | - |
mutant enzyme E132D, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
0.3 | - |
mutant enzyme W13A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
0.6 | - |
mutant enzyme E67Q, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
0.74 | - |
wild type enzyme, using ribulose 5-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
1.4 | - |
mutant enzyme E132A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
1.5 | - |
mutant enzyme H122Q, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
1.5 | - |
mutant enzyme H50Q, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
1.5 | - |
mutant enzyme Q48A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
1.7 | - |
mutant enzyme H50A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
1.9 | - |
mutant enzyme E67A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
1.9 | - |
mutant enzyme R142A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
2.1 | - |
mutant enzyme H122A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
2.8 | - |
mutant enzyme E67D, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
3 | 8 | mutant enzyme W69A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
4.5 | - |
wild type enzyme, using D-fructose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
11 | - |
mutant enzyme K65A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
16 | - |
mutant enzyme K37A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
31 | - |
mutant enzyme W13F, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
39 | - |
mutant enzyme W13H, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
39 | - |
mutant enzyme Y124A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
40 | - |
mutant enzyme L39A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
44 | - |
mutant enzyme D138A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
51 | - |
mutant enzyme W13Y, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
58 | - |
mutant enzyme L18A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
75 | - |
wild type enzyme, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
194 | - |
mutant enzyme R11A, using D-mannose 6-phosphate as substrate, at pH 7.0 and 80°C | Thermus thermophilus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-fructose 6-phosphate | - |
Thermus thermophilus | D-mannose 6-phosphate | - |
r | |
D-fructose 6-phosphate | - |
Thermus thermophilus KCCM 40879 | D-mannose 6-phosphate | - |
r | |
D-Mannose 6-phosphate | best substrate | Thermus thermophilus | D-Fructose 6-phosphate | - |
r | |
D-Mannose 6-phosphate | best substrate | Thermus thermophilus KCCM 40879 | D-Fructose 6-phosphate | - |
r | |
D-ribose 5-phosphate | very low activity | Thermus thermophilus | D-ribulose 5-phosphate | - |
r | |
D-ribose 5-phosphate | very low activity | Thermus thermophilus KCCM 40879 | D-ribulose 5-phosphate | - |
r | |
additional information | no activity with D-glucose 6-phosphate and arabinose 5-phosphate | Thermus thermophilus | ? | - |
? | |
additional information | no activity with D-glucose 6-phosphate and arabinose 5-phosphate | Thermus thermophilus KCCM 40879 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 29000, SDS-PAGE | Thermus thermophilus |
monomer | 1 * 29054, calculated from amino acid sequence | Thermus thermophilus |
Synonyms | Comment | Organism |
---|---|---|
mannose-6-phosphate isomerase | - |
Thermus thermophilus |
MPI | - |
Thermus thermophilus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
80 | - |
- |
Thermus thermophilus |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
70 | 90 | - |
Thermus thermophilus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
64 | 85 | the half-lives of the enzyme at 65, 70, 75, 80, and 85°C are 13, 6.5, 3.7, 1.8, and 0.2 h, respectively | Thermus thermophilus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
11 | - |
D-mannose 6-phosphate | mutant enzyme W13A, pH 7.0, 80°C | Thermus thermophilus | |
19 | - |
D-mannose 6-phosphate | mutant enzyme E132A, pH 7.0, 80°C | Thermus thermophilus | |
31 | - |
D-mannose 6-phosphate | mutant enzyme E67A, pH 7.0, 80°C | Thermus thermophilus | |
37 | - |
D-mannose 6-phosphate | mutant enzyme Q48A, pH 7.0, 80°C | Thermus thermophilus | |
54 | - |
D-mannose 6-phosphate | mutant enzyme H50A, pH 7.0, 80°C | Thermus thermophilus | |
70 | - |
D-mannose 6-phosphate | mutant enzyme R142A, pH 7.0, 80°C | Thermus thermophilus | |
75 | - |
D-mannose 6-phosphate | mutant enzyme H122A, pH 7.0, 80°C | Thermus thermophilus | |
141 | - |
D-fructose 6-phosphate | wild type enzyme, pH 7.0, 80°C | Thermus thermophilus | |
1371 | - |
D-mannose 6-phosphate | wild type enzyme, pH 7.0, 80°C | Thermus thermophilus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
- |
Thermus thermophilus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6.5 | 8.5 | - |
Thermus thermophilus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
13 | - |
D-mannose 6-phosphate | mutant enzyme W13A, pH 7.0, 80°C | Thermus thermophilus | |
54 | - |
D-mannose 6-phosphate | mutant enzyme R142A, pH 7.0, 80°C | Thermus thermophilus | |
69 | - |
D-mannose 6-phosphate | mutant enzyme Q48A, pH 7.0, 80°C | Thermus thermophilus | |
109 | - |
D-mannose 6-phosphate | mutant enzyme E132A, pH 7.0, 80°C | Thermus thermophilus | |
166 | - |
D-mannose 6-phosphate | mutant enzyme H50A, pH 7.0, 80°C | Thermus thermophilus | |
174 | - |
D-mannose 6-phosphate | mutant enzyme E67A, pH 7.0, 80°C | Thermus thermophilus | |
278 | - |
D-mannose 6-phosphate | mutant enzyme H122A, pH 7.0, 80°C | Thermus thermophilus | |
641 | - |
D-fructose 6-phosphate | wild type enzyme, pH 7.0, 80°C | Thermus thermophilus | |
6685 | - |
D-mannose 6-phosphate | wild type enzyme, pH 7.0, 80°C | Thermus thermophilus |