Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 2.2.1.1 extracted from

  • Hibbert, E.G.; Senussi, T.; Costelloe, S.J.; Lei, W.; Smith, M.E.; Ward, J.M.; Hailes, H.C.; Dalby, P.A.
    Directed evolution of transketolase activity on non-phosphorylated substrates (2007), J. Biotechnol., 131, 425-432.
    View publication on PubMed
Search for more literature for 2.2.1.1

Application

Application Comment Organism
additional information phylogenetically variant active-site residues are useful for modulating activity of transketolase on natural or structurally-homologous substrates, whereas conserved residues which no longer interact with modified target substrates are useful sites to apply saturation mutagenesis for improvement of activity of transketolase Escherichia coli

Cloned(Commentary)

Cloned (Comment) Organism
tkta gene complete with its natural promoter on the pQR711 plasmid expressed in Escherichia coli TOP10 or XL10 cells. A29E mutation introduced into the pQR412 vector, in which the transketolase gene has an N-terminal His×6 tag. Both wild-type and mutant A29E overexpressed from pQR412 Escherichia coli

Protein Variants

Protein Variants Comment Organism
A29D specific activity is 2.7fold greater than for the wild-type Escherichia coli
A29E specific activity is 3fold greater than for the wild-type Escherichia coli
D259A specific activity is 2.3fold greater than for the wild-type Escherichia coli
D259G specific activity is 2fold greater than for the wild-type Escherichia coli
H461S specific activity is 4.8fold greater than for the wild-type Escherichia coli
R358I specific activity is 2.1fold greater than for the wild-type Escherichia coli
R358P specific activity is 1.5fold greater than for the wild-type Escherichia coli
R520G specific activity is 2.1fold greater than for the wild-type Escherichia coli
R520P specific activity is 2.3fold greater than for the wild-type Escherichia coli
R520Stop specific activity is 3.6fold greater than for the wild-type Escherichia coli
R520V specific activity is 3.6fold greater than for the wild-type Escherichia coli
S188Q specific activity is 2.3fold greater than for the wild-type Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5.3
-
 Hydroxypyruvate wild-type, in the presence of 50 mM glycolaldehyde Escherichia coli
6.6
-
 Hydroxypyruvate mutant A29E, in the presence of 50 mM glycolaldehyde Escherichia coli
35
-
 glycolaldehyde wild-type, in the presence of 50 mM hydroxypyruvate Escherichia coli
200
-
 glycolaldehyde mutant A29E, in the presence of 50 mM hydroxypyruvate Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Purification (Commentary)

Purification (Comment) Organism
wild-type and mutant A29E purified on Ni-NTA resin Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.65
-
 wild-type, in the presence of 50 mM hydroxypyruvate, 50 mM glycolaldehyde, and 50 mM Tris-HCl, 2.4 mM thiamine diphosphate, 9 mM MgCl2, pH 7.0 Escherichia coli
1
-
 mutant R358P, in the presence of 50 mM hydroxypyruvate, 50 mM glycolaldehyde, and 50 mM Tris-HCl, 2.4 mM thiamine diphosphate, 9 mM MgCl2, pH 7.0 Escherichia coli
1.3
-
 mutant D259G, in the presence of 50 mM hydroxypyruvate, 50 mM glycolaldehyde, and 50 mM Tris-HCl, 2.4 mM thiamine diphosphate, 9 mM MgCl2, pH 7.0 Escherichia coli
1.37
-
 mutant R358I, in the presence of 50 mM hydroxypyruvate, 50 mM glycolaldehyde, and 50 mM Tris-HCl, 2.4 mM thiamine diphosphate, 9 mM MgCl2, pH 7.0 Escherichia coli
1.4
-
 mutant R520G, in the presence of 50 mM hydroxypyruvate, 50 mM glycolaldehyde, and 50 mM Tris-HCl, 2.4 mM thiamine diphosphate, 9 mM MgCl2, pH 7.0 Escherichia coli
1.5
-
 mutant D259A, in the presence of 50 mM hydroxypyruvate, 50 mM glycolaldehyde, and 50 mM Tris-HCl, 2.4 mM thiamine diphosphate, 9 mM MgCl2, pH 7.0 Escherichia coli
1.5
-
 mutant R520P, in the presence of 50 mM hydroxypyruvate, 50 mM glycolaldehyde, and 50 mM Tris-HCl, 2.4 mM thiamine diphosphate, 9 mM MgCl2, pH 7.0 Escherichia coli
1.5
-
 mutant S188Q, in the presence of 50 mM hydroxypyruvate, 50 mM glycolaldehyde, and 50 mM Tris-HCl, 2.4 mM thiamine diphosphate, 9 mM MgCl2, pH 7.0 Escherichia coli
1.8
-
 mutant A29D, in the presence of 50 mM hydroxypyruvate, 50 mM glycolaldehyde, and 50 mM Tris-HCl, 2.4 mM thiamine diphosphate, 9 mM MgCl2, pH 7.0 Escherichia coli
1.95
-
 mutant A29E, in the presence of 50 mM hydroxypyruvate, 50 mM glycolaldehyde, and 50 mM Tris-HCl, 2.4 mM thiamine diphosphate, 9 mM MgCl2, pH 7.0 Escherichia coli
2.3
-
 mutant R520Stop, in the presence of 50 mM hydroxypyruvate, 50 mM glycolaldehyde, and 50 mM Tris-HCl, 2.4 mM thiamine diphosphate, 9 mM MgCl2, pH 7.0 Escherichia coli
2.3
-
 mutant R520V, in the presence of 50 mM hydroxypyruvate, 50 mM glycolaldehyde, and 50 mM Tris-HCl, 2.4 mM thiamine diphosphate, 9 mM MgCl2, pH 7.0 Escherichia coli
3.14
-
 mutant H461S, in the presence of 50 mM hydroxypyruvate, 50 mM glycolaldehyde, and 50 mM Tris-HCl, 2.4 mM thiamine diphosphate, 9 mM MgCl2, pH 7.0 Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
hydroxypyruvate + glycolaldehyde
-
 Escherichia coli L-erythrulose + ?
-
 ?

Synonyms

Synonyms Comment Organism
TktA
-
 Escherichia coli

Cofactor

Cofactor Comment Organism Structure
thiamine diphosphate
-
 Escherichia coli