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Literature summary for 5.3.1.4 extracted from

  • Hung, X.; Tseng, W.; Liu, S.; Tzou, W.; Fang, T.
    Characterization of a thermophilic L-arabinose isomerase from Thermoanaerobacterium saccharolyticum NTOU1 (2014), Biochem. Eng. J., 83, 121-128.
No PubMed abstract available

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21-CodonPlus (DE3)-RIL cells Thermoanaerobacterium saccharolyticum

Inhibitors

Inhibitors Comment Organism Structure
Dulcitol 90% residual activity at 10 mM Thermoanaerobacterium saccharolyticum
erythritol 42% residual activity at 10 mM Thermoanaerobacterium saccharolyticum
L-arabitol 11% residual activity at 10 mM Thermoanaerobacterium saccharolyticum
ribitol 22% residual activity at 10 mM Thermoanaerobacterium saccharolyticum
xylitol 46% residual activity at 10 mM Thermoanaerobacterium saccharolyticum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
122
-
D-galactose in 25 mM Tris-HCl buffer (pH 7.5), at 60°C Thermoanaerobacterium saccharolyticum

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ 0.05 mM Co2+, can improve both catalytic activity and thermostability at higher temperatures Thermoanaerobacterium saccharolyticum
Mn2+ 0.1 mM Mn2+,increases activity to 167.3% and can improve both catalytic activity and thermostability at higher temperatures Thermoanaerobacterium saccharolyticum

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
52863
-
4 * 52863, native enzyme, calculated from amino acid sequence Thermoanaerobacterium saccharolyticum
55000
-
4 * 55000, recombinant enzyme, SDS-PAGE Thermoanaerobacterium saccharolyticum
55343
-
4 * 55343, recombinant enzyme, calculated from amino acid sequence Thermoanaerobacterium saccharolyticum
55397
-
4 * 55397, recombinant enzyme, MALDI-TOF mass spectrometry Thermoanaerobacterium saccharolyticum
233000
-
gel filtration Thermoanaerobacterium saccharolyticum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-fucose Thermoanaerobacterium saccharolyticum 221% activity compared to D-galactose D-fuculose
-
r
D-galactose Thermoanaerobacterium saccharolyticum 100% activity D-tagatose
-
r
L-arabinose Thermoanaerobacterium saccharolyticum 244% activity compared to D-galactose L-ribulose
-
r
L-arabinose Thermoanaerobacterium saccharolyticum NTOU1 244% activity compared to D-galactose L-ribulose
-
r

Organism

Organism UniProt Comment Textmining
Thermoanaerobacterium saccharolyticum K7SW59
-
-
Thermoanaerobacterium saccharolyticum NTOU1 K7SW59
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography Thermoanaerobacterium saccharolyticum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-Allose 9% activity compared to D-galactose Thermoanaerobacterium saccharolyticum ?
-
?
D-Allose 9% activity compared to D-galactose Thermoanaerobacterium saccharolyticum NTOU1 ?
-
?
D-fucose 221% activity compared to D-galactose Thermoanaerobacterium saccharolyticum D-fuculose
-
r
D-galactose 100% activity Thermoanaerobacterium saccharolyticum D-tagatose
-
r
D-rhamnose 28% activity compared to D-galactose Thermoanaerobacterium saccharolyticum ?
-
?
D-Ribose 7% activity compared to D-galactose Thermoanaerobacterium saccharolyticum ?
-
?
D-Ribose 7% activity compared to D-galactose Thermoanaerobacterium saccharolyticum NTOU1 ?
-
?
D-sorbose 31% activity compared to D-galactose Thermoanaerobacterium saccharolyticum ?
-
?
D-Xylose 14% activity compared to D-galactose Thermoanaerobacterium saccharolyticum ?
-
?
D-Xylose 14% activity compared to D-galactose Thermoanaerobacterium saccharolyticum NTOU1 ?
-
?
L-arabinose 244% activity compared to D-galactose Thermoanaerobacterium saccharolyticum L-ribulose
-
r
L-arabinose 244% activity compared to D-galactose Thermoanaerobacterium saccharolyticum NTOU1 L-ribulose
-
r
additional information no activity with D-glucose, D-mannose, and L-fucose Thermoanaerobacterium saccharolyticum ?
-
?
additional information no activity with D-glucose, D-mannose, and L-fucose Thermoanaerobacterium saccharolyticum NTOU1 ?
-
?

Subunits

Subunits Comment Organism
homotetramer 4 * 55000, recombinant enzyme, SDS-PAGE Thermoanaerobacterium saccharolyticum
homotetramer 4 * 52863, native enzyme, calculated from amino acid sequence Thermoanaerobacterium saccharolyticum
homotetramer 4 * 55343, recombinant enzyme, calculated from amino acid sequence Thermoanaerobacterium saccharolyticum
homotetramer 4 * 55397, recombinant enzyme, MALDI-TOF mass spectrometry Thermoanaerobacterium saccharolyticum

Synonyms

Synonyms Comment Organism
AraA
-
Thermoanaerobacterium saccharolyticum
L-AI
-
Thermoanaerobacterium saccharolyticum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
70
-
-
Thermoanaerobacterium saccharolyticum

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
40 80 the activity increases with the increasing temperature (starting with 22% activity at 40°C, 50% activity at 50°C), reaches an optimal activity at 70°C, and then drops with a steep slope down (65% activity at 75°C) to inactivity at 80°C Thermoanaerobacterium saccharolyticum

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
55 75 the activity is fully retained after 2 h incubation at 55-65°C. In the presence of 0.1 mM Mn2+ and 0.05 mM Co2+, the enzyme can remain stable for 2 h during the incubation at 55-65°C, and the half-life of the enzyme is about 150 and 15 min at 70 and 75°C, respectively Thermoanaerobacterium saccharolyticum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.9
-
D-galactose in 25 mM Tris-HCl buffer (pH 7.5), at 60°C Thermoanaerobacterium saccharolyticum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7 7.5
-
Thermoanaerobacterium saccharolyticum

pH Range

pH Minimum pH Maximum Comment Organism
5 9.5 very minimal activity when the pH either drops below 5.0 or increases to above 9.5 Thermoanaerobacterium saccharolyticum

pH Stability

pH Stability pH Stability Maximum Comment Organism
5.5 10.5 remains stable after 24 h. The enzyme activity is fully retained at pH 6.5-9.5 after 6 h with about 70% remaining at pH 5.5 and 10.5 Thermoanaerobacterium saccharolyticum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.04
-
D-galactose in 25 mM Tris-HCl buffer (pH 7.5), at 60°C Thermoanaerobacterium saccharolyticum