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Literature summary 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

Cloned(Commentary)

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

Protein Variants

EC Number Protein Variants Comment Organism
5.3.1.25 D349A inactive Dictyoglomus turgidum
5.3.1.25 D349D inactive Dictyoglomus turgidum
5.3.1.25 D349K inactive Dictyoglomus turgidum
5.3.1.25 D349Q inactive Dictyoglomus turgidum
5.3.1.25 D349W inactive Dictyoglomus turgidum
5.3.1.25 D373A the mutation results in a substantial increase in Km value Dictyoglomus turgidum
5.3.1.25 D373E inactive Dictyoglomus turgidum
5.3.1.25 D373K inactive Dictyoglomus turgidum
5.3.1.25 D373Q inactive Dictyoglomus turgidum
5.3.1.25 D373W inactive Dictyoglomus turgidum
5.3.1.25 F452A the mutation results in decreases in kcat, but has little effect on Km value Dictyoglomus turgidum
5.3.1.25 I199A the mutation results in a substantial increase in Km value Dictyoglomus turgidum
5.3.1.25 M197A the mutation results in a substantial increase in Km value Dictyoglomus turgidum
5.3.1.25 N404A the mutation results in decreases in kcat, but has little effect on Km value Dictyoglomus turgidum
5.3.1.25 N538A the mutation results in a substantial increase in Km value Dictyoglomus turgidum
5.3.1.25 Q314A the mutation results in a substantial increase in Km value Dictyoglomus turgidum
5.3.1.25 R30A the mutation results in decreases in kcat, but has little effect on Km value Dictyoglomus turgidum
5.3.1.25 S405A the mutation results in a substantial increase in Km value Dictyoglomus turgidum
5.3.1.25 V131A the mutation results in a substantial increase in Km value Dictyoglomus turgidum
5.3.1.25 W102A the mutation results in decreases in kcat, but has little effect on Km value Dictyoglomus turgidum
5.3.1.25 W510A the mutation results in decreases in kcat, but has little effect on Km value Dictyoglomus turgidum
5.3.1.25 Y451A the mutation results in a substantial increase in Km value Dictyoglomus turgidum

Inhibitors

EC Number Inhibitors Comment Organism Structure
5.3.1.25 Cu2+ inhibitory at 1 mM Dictyoglomus turgidum
5.3.1.25 EDTA complete inhibition at 1 mM Dictyoglomus turgidum
5.3.1.25 Fe2+ inhibitory at 1 mM Dictyoglomus turgidum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5.3.1.25 52
-
 L-fucose mutant enzyme H539A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 72
-
 L-fucose wild type enzyme, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 77
-
 L-fucose mutant enzyme W102A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 85
-
 L-fucose mutant enzyme R30A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 86
-
 L-fucose mutant enzyme F452A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 90
-
 L-fucose mutant enzyme W510A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 92
-
 L-fucose mutant enzyme N404A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 140
-
 L-fucose mutant enzyme D373A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 148
-
 L-fucose mutant enzyme V131A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 166
-
 L-fucose mutant enzyme M197A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 202
-
 L-fucose mutant enzyme I199A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 225
-
 L-fucose mutant enzyme Q314A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 302
-
 L-fucose mutant enzyme Y451A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 339
-
 L-fucose mutant enzyme N538A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 385
-
 L-fucose mutant enzyme S405A, at pH 7.0 and 80°C Dictyoglomus turgidum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
5.3.1.25 Co2+ about 5fold stimulation of activity at 1 mM Dictyoglomus turgidum
5.3.1.25 Mg2+ about 2fold stimulation of activity at 1 mM Dictyoglomus turgidum
5.3.1.25 Mn2+ required for maximum activity (more than 10fold stimulation of activity at 1 mM), the enzyme contains one atom per monomer Dictyoglomus turgidum
5.3.1.25 additional information Mg2+ and Ca2+ show no crucial effect on the enzyme activity Dictyoglomus turgidum
5.3.1.25 Zn2+ about 3fold stimulation of activity at 1 mM Dictyoglomus turgidum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
5.3.1.25 68000
-
 6 * 68000, His6-tagged enzyme, SDS-PAGE Dictyoglomus turgidum
5.3.1.25 68363
-
 6 * 68363, His6-tagged enzyme, calculated from amino acid sequence Dictyoglomus turgidum
5.3.1.25 410000
-
 gel filtration Dictyoglomus turgidum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
5.3.1.25 D-altrose Dictyoglomus turgidum
-
 D-psicose
-
 ?
5.3.1.25 D-altrose Dictyoglomus turgidum DSM 6724
-
 D-psicose
-
 ?
5.3.1.25 D-arabinose Dictyoglomus turgidum
-
 D-ribose
-
 ?
5.3.1.25 D-arabinose Dictyoglomus turgidum DSM 6724
-
 D-ribose
-
 ?
5.3.1.25 L-Fucose Dictyoglomus turgidum highest activity L-Fuculose
-
 ?
5.3.1.25 L-Fucose Dictyoglomus turgidum DSM 6724 highest activity L-Fuculose
-
 ?
5.3.1.25 L-galactose Dictyoglomus turgidum
-
 L-tagatose
-
 ?
5.3.1.25 L-galactose Dictyoglomus turgidum DSM 6724
-
 L-tagatose
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
5.3.1.25 Dictyoglomus turgidum B8E1T1
-
-
5.3.1.25 Dictyoglomus turgidum DSM 6724 B8E1T1
-
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
5.3.1.25 0.35
-
 mutant enzyme W510A, using L-fucose as substrate, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 0.47
-
 mutant enzyme D373A, using L-fucose as substrate, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 0.67
-
 mutant enzyme Y451A, using L-fucose as substrate, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 0.93
-
 mutant enzyme Q314A, using L-fucose as substrate, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 1.3
-
 mutant enzyme N404A, using L-fucose as substrate, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 2.1
-
 mutant enzyme H539A, using L-fucose as substrate, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 2.4
-
 mutant enzyme M197A, using L-fucose as substrate, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 2.5
-
 mutant enzyme W102A, using L-fucose as substrate, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 2.6
-
 mutant enzyme F452A, using L-fucose as substrate, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 3.4
-
 mutant enzyme R30A, using L-fucose as substrate, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 4.3
-
 mutant enzyme I199A, using L-fucose as substrate, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 6.8
-
 mutant enzyme N538A, using L-fucose as substrate, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 27
-
 mutant enzyme V131A, using L-fucose as substrate, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 30
-
 mutant enzyme S405A, using L-fucose as substrate, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 33
-
 wild type enzyme, using L-fucose as substrate, at pH 7.0 and 80°C Dictyoglomus turgidum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.3.1.25 D-altrose
-
 Dictyoglomus turgidum D-psicose
-
 ?
5.3.1.25 D-altrose
-
 Dictyoglomus turgidum DSM 6724 D-psicose
-
 ?
5.3.1.25 D-arabinose
-
 Dictyoglomus turgidum D-ribose
-
 ?
5.3.1.25 D-arabinose
-
 Dictyoglomus turgidum DSM 6724 D-ribose
-
 ?
5.3.1.25 L-Fucose highest activity Dictyoglomus turgidum L-Fuculose
-
 ?
5.3.1.25 L-Fucose highest activity Dictyoglomus turgidum DSM 6724 L-Fuculose
-
 ?
5.3.1.25 L-galactose
-
 Dictyoglomus turgidum L-tagatose
-
 ?
5.3.1.25 L-galactose
-
 Dictyoglomus turgidum DSM 6724 L-tagatose
-
 ?

Subunits

EC Number Subunits Comment Organism
5.3.1.25 homohexamer 6 * 68000, His6-tagged enzyme, SDS-PAGE Dictyoglomus turgidum
5.3.1.25 homohexamer 6 * 68363, His6-tagged enzyme, calculated from amino acid sequence Dictyoglomus turgidum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
5.3.1.25 80
-
-
 Dictyoglomus turgidum

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
5.3.1.25 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]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
5.3.1.25 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]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.3.1.25 2.58
-
 L-fucose mutant enzyme W510A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 3.37
-
 L-fucose mutant enzyme H539A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 7.38
-
 L-fucose mutant enzyme Q314A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 8
-
 L-fucose mutant enzyme D373A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 10.37
-
 L-fucose mutant enzyme N404A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 17.83
-
 L-fucose mutant enzyme M197A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 19.17
-
 L-fucose mutant enzyme W102A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 20.33
-
 L-fucose mutant enzyme F452A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 33.33
-
 L-fucose mutant enzyme I199A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 41
-
 L-fucose mutant enzyme Y451A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 53.83
-
 L-fucose mutant enzyme R30A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 114.8
-
 L-fucose mutant enzyme N538A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 216.7
-
 L-fucose mutant enzyme V131A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 235
-
 L-fucose mutant enzyme S405A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 258.3
-
 L-fucose wild type enzyme, at pH 7.0 and 80°C Dictyoglomus turgidum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
5.3.1.25 7
-
-
 Dictyoglomus turgidum

pH Range

EC Number pH Minimum pH Maximum Comment Organism
5.3.1.25 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]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5.3.1.25 0.03
-
 L-fucose mutant enzyme Q314A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 0.03
-
 L-fucose mutant enzyme W510A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 0.06
-
 L-fucose mutant enzyme D373A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 0.07
-
 L-fucose mutant enzyme H539A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 0.11
-
 L-fucose mutant enzyme M197A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 0.11
-
 L-fucose mutant enzyme N404A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 0.14
-
 L-fucose mutant enzyme Y451A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 0.16
-
 L-fucose mutant enzyme I199A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 0.23
-
 L-fucose mutant enzyme F452A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 0.25
-
 L-fucose mutant enzyme W102A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 0.33
-
 L-fucose mutant enzyme N538A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 0.62
-
 L-fucose mutant enzyme R30A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 0.62
-
 L-fucose mutant enzyme S405A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 1.48
-
 L-fucose mutant enzyme V131A, at pH 7.0 and 80°C Dictyoglomus turgidum
5.3.1.25 3.58
-
 L-fucose wild type enzyme, at pH 7.0 and 80°C Dictyoglomus turgidum