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Literature summary for 5.3.1.25 extracted from

  • Hong, S.H.; Lim, Y.R.; Kim, Y.S.; Oh, D.K.
    Molecular characterization of a thermostable L-fucose isomerase from Dictyoglomus turgidum that isomerizes L-fucose and D-arabinose (2012), Biochimie, 94, 1926-1934.
    View publication on PubMed
Search for more literature for 5.3.1.25

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli ER2566 cells Dictyoglomus turgidum

Protein Variants

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

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 [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

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 [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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Dictyoglomus turgidum B8E1T1
-
-
Dictyoglomus turgidum DSM 6724 B8E1T1
-
-

Purification (Commentary)

Purification (Comment) Organism
heat treatment and His-Trap affinity chromatography Dictyoglomus turgidum

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
80
-
-
 Dictyoglomus turgidum

Temperature Range [°C]

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 [°C]

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 [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
-
 Dictyoglomus turgidum

pH Range

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 [mM/s]

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