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Literature summary for 1.1.1.25 extracted from

  • Garcia-Guevara, F.; Bravo, I.; Martinez-Anaya, C.; Segovia, L.
    Cofactor specificity switch in shikimate dehydrogenase by rational design and consensus engineering (2017), Protein Eng. Des. Sel., 30, 533-541 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene aroE, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli Escherichia coli

Protein Variants

Protein Variants Comment Organism
A243G site-directed mutagenesis, the mutant shows altered cofactor specificity compared to wild-type enzyme Escherichia coli
D195E site-directed mutagenesis, the mutant shows altered cofactor specificity compared to wild-type enzyme Escherichia coli
additional information invertion of the cofactor specificity from NADP+ to NAD+ on the Escherichia coli wild-type enzyme, effect of consensus mutations, overview. Mutant structure modeling Escherichia coli
N149D site-directed mutagenesis, the mutant shows altered cofactor specificity compared to wild-type enzyme Escherichia coli
N149D/V152F site-directed mutagenesis, the mutant shows altered cofactor specificity compared to wild-type enzyme Escherichia coli
S131A site-directed mutagenesis, the mutant shows altered cofactor specificity compared to wild-type enzyme Escherichia coli
S131A/L135A site-directed mutagenesis, the mutant shows altered cofactor specificity compared to wild-type enzyme Escherichia coli
S131A/L135A/N149D/V152F site-directed mutagenesis, the mutant shows altered cofactor specificity compared to wild-type enzyme Escherichia coli
S131A/N149D/V152F site-directed mutagenesis, the mutant shows altered cofactor specificity compared to wild-type enzyme Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information Michaelis-Menten kinetics Escherichia coli
0.13
-
shikimate pH 9.0, 25°C, recombinant wild-type enzyme Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
shikimate + NAD+ Escherichia coli
-
3-dehydroshikimate + NADH + H+
-
r
shikimate + NADP+ Escherichia coli
-
3-dehydroshikimate + NADPH + H+
-
r

Organism

Organism UniProt Comment Textmining
Escherichia coli P15770
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography and dialysis Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
shikimate + NAD+
-
Escherichia coli 3-dehydroshikimate + NADH + H+
-
r
shikimate + NADP+
-
Escherichia coli 3-dehydroshikimate + NADPH + H+
-
r

Synonyms

Synonyms Comment Organism
AroE
-
Escherichia coli
EcoSDH
-
Escherichia coli
SDH
-
Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
3-dehydroshikimate formation, assay at Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
190
-
shikimate pH 9.0, 25°C, recombinant wild-type enzyme Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
3-dehydroshikimate formation, assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Escherichia coli
NADH
-
Escherichia coli
NADP+ preferred cofactor of the wild-type enzyme Escherichia coli
NADPH
-
Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.322
-
NADP+ pH 9.0, 25°C, recombinant mutant N149D Escherichia coli
0.457
-
NADP+ pH 9.0, 25°C, recombinant mutant N149D/V152F Escherichia coli
1.42
-
NAD+ pH 9.0, 25°C, recombinant wild-type enzyme Escherichia coli
1.8
-
NAD+ pH 9.0, 25°C, recombinant mutant N149D Escherichia coli
2.217
-
NADP+ pH 9.0, 25°C, recombinant mutant S131A/N149D/V152F Escherichia coli
3.133
-
NADP+ pH 9.0, 25°C, recombinant mutant S131A/L135A/N149D/V152F Escherichia coli
4.17
-
NAD+ pH 9.0, 25°C, recombinant mutant N149D/V152F Escherichia coli
12.55
-
NAD+ pH 9.0, 25°C, recombinant mutant S131A/N149D/V152F Escherichia coli
16.72
-
NAD+ pH 9.0, 25°C, recombinant mutant S131A/L135A/N149D/V152F Escherichia coli
1461.5
-
shikimate pH 9.0, 25°C, recombinant wild-type enzyme Escherichia coli
3200
-
NADP+ pH 9.0, 25°C, recombinant wild-type enzyme Escherichia coli