Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli ER2566 cells | Dictyoglomus turgidum |
Protein Variants | Comment | Organism |
---|---|---|
D349A | inactive | Dictyoglomus turgidum |
D349D | inactive | Dictyoglomus turgidum |
D349K | inactive | Dictyoglomus turgidum |
D349Q | inactive | Dictyoglomus turgidum |
D349W | inactive | Dictyoglomus turgidum |
D373A | the mutation results in a substantial increase in Km value | Dictyoglomus turgidum |
D373E | inactive | Dictyoglomus turgidum |
D373K | inactive | Dictyoglomus turgidum |
D373Q | inactive | Dictyoglomus turgidum |
D373W | inactive | Dictyoglomus turgidum |
F452A | the mutation results in decreases in kcat, but has little effect on Km value | Dictyoglomus turgidum |
I199A | the mutation results in a substantial increase in Km value | Dictyoglomus turgidum |
M197A | the mutation results in a substantial increase in Km value | Dictyoglomus turgidum |
N404A | the mutation results in decreases in kcat, but has little effect on Km value | Dictyoglomus turgidum |
N538A | the mutation results in a substantial increase in Km value | Dictyoglomus turgidum |
Q314A | the mutation results in a substantial increase in Km value | Dictyoglomus turgidum |
R30A | the mutation results in decreases in kcat, but has little effect on Km value | Dictyoglomus turgidum |
S405A | the mutation results in a substantial increase in Km value | Dictyoglomus turgidum |
V131A | the mutation results in a substantial increase in Km value | Dictyoglomus turgidum |
W102A | the mutation results in decreases in kcat, but has little effect on Km value | Dictyoglomus turgidum |
W510A | the mutation results in decreases in kcat, but has little effect on Km value | Dictyoglomus turgidum |
Y451A | the mutation results in a substantial increase in Km value | Dictyoglomus turgidum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Cu2+ | inhibitory at 1 mM | Dictyoglomus turgidum | |
EDTA | complete inhibition at 1 mM | Dictyoglomus turgidum | |
Fe2+ | inhibitory at 1 mM | Dictyoglomus turgidum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
52 | - |
L-fucose | mutant enzyme H539A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
72 | - |
L-fucose | wild type enzyme, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
77 | - |
L-fucose | mutant enzyme W102A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
85 | - |
L-fucose | mutant enzyme R30A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
86 | - |
L-fucose | mutant enzyme F452A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
90 | - |
L-fucose | mutant enzyme W510A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
92 | - |
L-fucose | mutant enzyme N404A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
140 | - |
L-fucose | mutant enzyme D373A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
148 | - |
L-fucose | mutant enzyme V131A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
166 | - |
L-fucose | mutant enzyme M197A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
202 | - |
L-fucose | mutant enzyme I199A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
225 | - |
L-fucose | mutant enzyme Q314A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
302 | - |
L-fucose | mutant enzyme Y451A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
339 | - |
L-fucose | mutant enzyme N538A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
385 | - |
L-fucose | mutant enzyme S405A, at pH 7.0 and 80°C | Dictyoglomus turgidum |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | about 5fold stimulation of activity at 1 mM | Dictyoglomus turgidum | |
Mg2+ | about 2fold stimulation of activity at 1 mM | Dictyoglomus turgidum | |
Mn2+ | required for maximum activity (more than 10fold stimulation of activity at 1 mM), the enzyme contains one atom per monomer | Dictyoglomus turgidum | |
additional information | Mg2+ and Ca2+ show no crucial effect on the enzyme activity | Dictyoglomus turgidum | |
Zn2+ | about 3fold stimulation of activity at 1 mM | Dictyoglomus turgidum |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
68000 | - |
6 * 68000, His6-tagged enzyme, SDS-PAGE | Dictyoglomus turgidum |
68363 | - |
6 * 68363, His6-tagged enzyme, calculated from amino acid sequence | Dictyoglomus turgidum |
410000 | - |
gel filtration | Dictyoglomus turgidum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-altrose | Dictyoglomus turgidum | - |
D-psicose | - |
? | |
D-altrose | Dictyoglomus turgidum DSM 6724 | - |
D-psicose | - |
? | |
D-arabinose | Dictyoglomus turgidum | - |
D-ribose | - |
? | |
D-arabinose | Dictyoglomus turgidum DSM 6724 | - |
D-ribose | - |
? | |
L-Fucose | Dictyoglomus turgidum | highest activity | L-Fuculose | - |
? | |
L-Fucose | Dictyoglomus turgidum DSM 6724 | highest activity | L-Fuculose | - |
? | |
L-galactose | Dictyoglomus turgidum | - |
L-tagatose | - |
? | |
L-galactose | Dictyoglomus turgidum DSM 6724 | - |
L-tagatose | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Dictyoglomus turgidum | B8E1T1 | - |
- |
Dictyoglomus turgidum DSM 6724 | B8E1T1 | - |
- |
Purification (Comment) | Organism |
---|---|
heat treatment and His-Trap affinity chromatography | Dictyoglomus turgidum |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.35 | - |
mutant enzyme W510A, using L-fucose as substrate, at pH 7.0 and 80°C | Dictyoglomus turgidum |
0.47 | - |
mutant enzyme D373A, using L-fucose as substrate, at pH 7.0 and 80°C | Dictyoglomus turgidum |
0.67 | - |
mutant enzyme Y451A, using L-fucose as substrate, at pH 7.0 and 80°C | Dictyoglomus turgidum |
0.93 | - |
mutant enzyme Q314A, using L-fucose as substrate, at pH 7.0 and 80°C | Dictyoglomus turgidum |
1.3 | - |
mutant enzyme N404A, using L-fucose as substrate, at pH 7.0 and 80°C | Dictyoglomus turgidum |
2.1 | - |
mutant enzyme H539A, using L-fucose as substrate, at pH 7.0 and 80°C | Dictyoglomus turgidum |
2.4 | - |
mutant enzyme M197A, using L-fucose as substrate, at pH 7.0 and 80°C | Dictyoglomus turgidum |
2.5 | - |
mutant enzyme W102A, using L-fucose as substrate, at pH 7.0 and 80°C | Dictyoglomus turgidum |
2.6 | - |
mutant enzyme F452A, using L-fucose as substrate, at pH 7.0 and 80°C | Dictyoglomus turgidum |
3.4 | - |
mutant enzyme R30A, using L-fucose as substrate, at pH 7.0 and 80°C | Dictyoglomus turgidum |
4.3 | - |
mutant enzyme I199A, using L-fucose as substrate, at pH 7.0 and 80°C | Dictyoglomus turgidum |
6.8 | - |
mutant enzyme N538A, using L-fucose as substrate, at pH 7.0 and 80°C | Dictyoglomus turgidum |
27 | - |
mutant enzyme V131A, using L-fucose as substrate, at pH 7.0 and 80°C | Dictyoglomus turgidum |
30 | - |
mutant enzyme S405A, using L-fucose as substrate, at pH 7.0 and 80°C | Dictyoglomus turgidum |
33 | - |
wild type enzyme, using L-fucose as substrate, at pH 7.0 and 80°C | Dictyoglomus turgidum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-altrose | - |
Dictyoglomus turgidum | D-psicose | - |
? | |
D-altrose | - |
Dictyoglomus turgidum DSM 6724 | D-psicose | - |
? | |
D-arabinose | - |
Dictyoglomus turgidum | D-ribose | - |
? | |
D-arabinose | - |
Dictyoglomus turgidum DSM 6724 | D-ribose | - |
? | |
L-Fucose | highest activity | Dictyoglomus turgidum | L-Fuculose | - |
? | |
L-Fucose | highest activity | Dictyoglomus turgidum DSM 6724 | L-Fuculose | - |
? | |
L-galactose | - |
Dictyoglomus turgidum | L-tagatose | - |
? | |
L-galactose | - |
Dictyoglomus turgidum DSM 6724 | L-tagatose | - |
? |
Subunits | Comment | Organism |
---|---|---|
homohexamer | 6 * 68000, His6-tagged enzyme, SDS-PAGE | Dictyoglomus turgidum |
homohexamer | 6 * 68363, His6-tagged enzyme, calculated from amino acid sequence | Dictyoglomus turgidum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
80 | - |
- |
Dictyoglomus turgidum |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
65 | 90 | about 50% activity at 65°C, about 60% activity at 70°C, about 75% activity at 75°C, 100% activity at 80°C, about 75% activity at 85°C, about 60% activity at 90°C | Dictyoglomus turgidum |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
80 | - |
the enzyme has half-lives of 20, 12, 7, 5, and 2 h at 65, 70,75, 80, and 85°C, respectively | Dictyoglomus turgidum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.58 | - |
L-fucose | mutant enzyme W510A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
3.37 | - |
L-fucose | mutant enzyme H539A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
7.38 | - |
L-fucose | mutant enzyme Q314A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
8 | - |
L-fucose | mutant enzyme D373A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
10.37 | - |
L-fucose | mutant enzyme N404A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
17.83 | - |
L-fucose | mutant enzyme M197A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
19.17 | - |
L-fucose | mutant enzyme W102A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
20.33 | - |
L-fucose | mutant enzyme F452A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
33.33 | - |
L-fucose | mutant enzyme I199A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
41 | - |
L-fucose | mutant enzyme Y451A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
53.83 | - |
L-fucose | mutant enzyme R30A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
114.8 | - |
L-fucose | mutant enzyme N538A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
216.7 | - |
L-fucose | mutant enzyme V131A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
235 | - |
L-fucose | mutant enzyme S405A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
258.3 | - |
L-fucose | wild type enzyme, at pH 7.0 and 80°C | Dictyoglomus turgidum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
- |
Dictyoglomus turgidum |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6.5 | 8.5 | about 80% activity at pH 6.5, 100% activity at pH 7.0, about 80% activity at pH 7.5, about 60% activity at pH 8.0, about 50% activity at pH 8.5 | Dictyoglomus turgidum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.03 | - |
L-fucose | mutant enzyme Q314A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
0.03 | - |
L-fucose | mutant enzyme W510A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
0.06 | - |
L-fucose | mutant enzyme D373A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
0.07 | - |
L-fucose | mutant enzyme H539A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
0.11 | - |
L-fucose | mutant enzyme M197A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
0.11 | - |
L-fucose | mutant enzyme N404A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
0.14 | - |
L-fucose | mutant enzyme Y451A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
0.16 | - |
L-fucose | mutant enzyme I199A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
0.23 | - |
L-fucose | mutant enzyme F452A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
0.25 | - |
L-fucose | mutant enzyme W102A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
0.33 | - |
L-fucose | mutant enzyme N538A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
0.62 | - |
L-fucose | mutant enzyme R30A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
0.62 | - |
L-fucose | mutant enzyme S405A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
1.48 | - |
L-fucose | mutant enzyme V131A, at pH 7.0 and 80°C | Dictyoglomus turgidum | |
3.58 | - |
L-fucose | wild type enzyme, at pH 7.0 and 80°C | Dictyoglomus turgidum |