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Literature summary for 2.5.1.55 extracted from

  • Taylor, W.P.; Sheflyan, G.Y.; Woodard, R.W.
    A single point mutation in 3-deoxy-D-manno-octulosonate-8-phosphate synthase is responsible for temperature sensitivity in a mutant strain of Salmonella typhimurium (2000), J. Biol. Chem., 275, 32141-32146.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
enzyme from wild-type strain AG701 and temperature-sensitive strain AG701i50 expressed in Escherichia coli Salmonella enterica subsp. enterica serovar Typhimurium

Protein Variants

Protein Variants Comment Organism
P145S natural mutation in the temperature-sensitive strain AG701i50 leads to an increase in Km-value for both substrates, D-arabinose 5-phosphate and phosphoenolpyruvate, this mutant enzyme also has an altered oligomeric state. Reduced activity, about 35% of the wild-type, between 15 and 30°C. Above 30°C the activity of the mutant enzyme decreases dramatically Salmonella enterica subsp. enterica serovar Typhimurium

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0028
-
phosphoenolpyruvate pH 7.4, 25°C, wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium
0.0042
-
phosphoenolpyruvate pH 7.4, 37°C, wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium
0.0044
-
D-arabinose 5-phosphate pH 7.4, 25°C, wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium
0.0093
-
D-arabinose 5-phosphate pH 7.4, 37°C, wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium
0.014
-
D-arabinose 5-phosphate pH 7.4, 25°C, mutant enzyme P145S Salmonella enterica subsp. enterica serovar Typhimurium
0.26
-
phosphoenolpyruvate pH 7.4, 25°C, mutant enzyme P145S Salmonella enterica subsp. enterica serovar Typhimurium
0.28
-
D-arabinose 5-phosphate pH 7.4, 37°C, mutant enzyme P145S Salmonella enterica subsp. enterica serovar Typhimurium
1.65
-
phosphoenolpyruvate pH 7.4, 37°C, mutant enzyme P145S Salmonella enterica subsp. enterica serovar Typhimurium

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
additional information
-
two poorly resolved peaks are detected by gel filtration of the mutant enzyme P145S: 91000 Da and 53000 Da Salmonella enterica subsp. enterica serovar Typhimurium
97000
-
wild-type enzyme, gel filtration Salmonella enterica subsp. enterica serovar Typhimurium

Organism

Organism UniProt Comment Textmining
Salmonella enterica subsp. enterica serovar Typhimurium
-
wild-type strain AG701 and temperature-sensitive strain AG701i50
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme from wild-type strain AG701 and temperature-sensitive strain AG701i50 Salmonella enterica subsp. enterica serovar Typhimurium

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
phosphoenolpyruvate + D-arabinose 5-phosphate
-
Salmonella enterica subsp. enterica serovar Typhimurium 2-dehydro-3-deoxy-D-octonate 8-phosphate + phosphate
-
?

Subunits

Subunits Comment Organism
trimer
-
Salmonella enterica subsp. enterica serovar Typhimurium

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
37
-
1 h, wild-type enzyme retains full activity, 50% loss of activity of the mutant enzyme P145S Salmonella enterica subsp. enterica serovar Typhimurium

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.72
-
phosphoenolpyruvate pH 7.4, 25°C, mutant enzyme P145S Salmonella enterica subsp. enterica serovar Typhimurium
0.74
-
D-arabinose 5-phosphate pH 7.4, 25°C, mutant enzyme P145S Salmonella enterica subsp. enterica serovar Typhimurium
1.63
-
D-arabinose 5-phosphate pH 7.4, 37°C, mutant enzyme P145S Salmonella enterica subsp. enterica serovar Typhimurium
1.75
-
D-arabinose 5-phosphate pH 7.4, 25°C, wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium
2.02
-
phosphoenolpyruvate pH 7.4, 37°C, mutant enzyme P145S Salmonella enterica subsp. enterica serovar Typhimurium
2.08
-
phosphoenolpyruvate pH 7.4, 25°C, wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium
5.6
-
D-arabinose 5-phosphate pH 7.4, 37°C, wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium
6.7
-
phosphoenolpyruvate pH 7.4, 37°C, wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium