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Literature summary for 5.3.1.4 extracted from

  • Lee, Y.J.; Lee, S.J.; Kim, S.B.; Lee, S.J.; Lee, S.H.; Lee, D.W.
    Structural insights into conserved L-arabinose metabolic enzymes reveal the substrate binding site of a thermophilic L-arabinose isomerase (2014), FEBS Lett., 588, 1064-1070.
    View publication on PubMed
Search for more literature for 5.3.1.4

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Geobacillus stearothermophilus

Protein Variants

Protein Variants Comment Organism
D118V the mutant shows 42% of wild type activity Geobacillus stearothermophilus
D195V the mutant shows 29% of wild type activity Geobacillus stearothermophilus
D309V the mutant shows 78% of wild type activity Geobacillus stearothermophilus
D333V the mutant shows 40% of wild type activity Geobacillus stearothermophilus
E133L the mutant shows 65% of wild type activity Geobacillus stearothermophilus
E233L the mutant shows wild type activity Geobacillus stearothermophilus
E261L the mutant shows 29% of wild type activity Geobacillus stearothermophilus
E332L the mutant shows 67% of wild type activity Geobacillus stearothermophilus
K196F the mutant shows 56% of wild type activity Geobacillus stearothermophilus
N406L the mutant shows 57% of wild type activity Geobacillus stearothermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
42.7
-
 L-arabinose mutant enzyme D118V, at pH 7.5 and 70°C Geobacillus stearothermophilus
57.9
-
 L-arabinose wild type enzyme, at pH 7.5 and 70°C Geobacillus stearothermophilus
80
-
 L-arabinose mutant enzyme N460L, at pH 7.5 and 70°C Geobacillus stearothermophilus
100
-
 L-arabinose mutant enzyme D333V, at pH 7.5 and 70°C Geobacillus stearothermophilus
119.3
-
 L-arabinose mutant enzyme D195V, at pH 7.5 and 70°C Geobacillus stearothermophilus
147.1
-
 L-arabinose mutant enzyme E261L, at pH 7.5 and 70°C Geobacillus stearothermophilus

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ about 2fold increase in activity at 1 mM Mn2+ Geobacillus stearothermophilus
additional information not significantly stimulated by Mg2+ Geobacillus stearothermophilus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-arabinose Geobacillus stearothermophilus
-
 L-ribulose
-
 ?

Organism

Organism UniProt Comment Textmining
Geobacillus stearothermophilus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
His-bind resin column chromatography, and Superdex 200 gel filtration Geobacillus stearothermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-arabinose
-
 Geobacillus stearothermophilus L-ribulose
-
 ?

Subunits

Subunits Comment Organism
homohexamer
-
 Geobacillus stearothermophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
11.2
-
 L-arabinose mutant enzyme E261L, at pH 7.5 and 70°C Geobacillus stearothermophilus
11.9
-
 L-arabinose mutant enzyme D118V, at pH 7.5 and 70°C Geobacillus stearothermophilus
14.2
-
 L-arabinose mutant enzyme D195V, at pH 7.5 and 70°C Geobacillus stearothermophilus
14.9
-
 L-arabinose mutant enzyme D333V, at pH 7.5 and 70°C Geobacillus stearothermophilus
27.8
-
 L-arabinose mutant enzyme N460L, at pH 7.5 and 70°C Geobacillus stearothermophilus
33.8
-
 L-arabinose wild type enzyme, at pH 7.5 and 70°C Geobacillus stearothermophilus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.08
-
 L-arabinose mutant enzyme E261L, at pH 7.5 and 70°C Geobacillus stearothermophilus
0.12
-
 L-arabinose mutant enzyme D195V, at pH 7.5 and 70°C Geobacillus stearothermophilus
0.15
-
 L-arabinose mutant enzyme D333V, at pH 7.5 and 70°C Geobacillus stearothermophilus
0.28
-
 L-arabinose mutant enzyme D118V, at pH 7.5 and 70°C Geobacillus stearothermophilus
0.35
-
 L-arabinose mutant enzyme N460L, at pH 7.5 and 70°C Geobacillus stearothermophilus
0.58
-
 L-arabinose wild type enzyme, at pH 7.5 and 70°C Geobacillus stearothermophilus