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

BRENDA support

Literature summary for 4.2.1.46 extracted from

  • Hegeman, A.D.; Gross, J.W.; Frey, P.A.
    Probing catalysis by Escherichia coli dTDP-glucose-4,6-dehydratase: identification and preliminary characterization of functional amino acid residues at the active site (2001), Biochemistry, 40, 6598-6610.
    View publication on PubMed
Search for more literature for 4.2.1.46

Protein Variants

Protein Variants Comment Organism
C187A 9.4fold decrease in turnover number for dTDP-glucose compared to wild-type value, 6fold decrease in KM-value for dTDP-glucose compared to wild-type value. 8% of the mutant enzyme contains NADH during steady-state turnover by adding a large excess of dTDPglucose, compared to 45% of the wild-type enzyme Escherichia coli
C187S 4.1fold decrease in turnover number for dTDP-glucose compared to wild-type value, 4fold decrease in KM-value for dTDP-glucose compared to wild-type value. 5% of the mutant enzyme contains NADH during steady-state turnover by adding a large excess of dTDPglucose, compared to 45% of the wild-type enzyme Escherichia coli
D135A 222fold decrease in turnover number for dTDP-glucose compared to wild-type value, 1.2fold increase in KM-value for dTDP-glucose compared to wild-type value. 30% of the mutant enzyme contains NADH during steady-state turnover by adding a large excess of dTDPglucose, compared to 45% of the wild-type enzyme Escherichia coli
D135N 124fold decrease in turnover number for dTDP-glucose compared to wild-type value, 1.3fold increase in KM-value for dTDP-glucose compared to wild-type value. 9% of the mutant enzyme contains NADH during steady-state turnover by adding a large excess of dTDPglucose, compared to 45% of the wild-type enzyme Escherichia coli
D135N/E136Q 204fold decrease in turnover number for dTDP-glucose compared to wild-type value, 3.2fold increase in KM-value for dTDP-glucose compared to wild-type value. Less than 0.5% of the mutant enzyme contains NADH during steady-state turnover by adding a large excess of dTDPglucose, compared to 45% of the wild-type enzyme Escherichia coli
E136A 288fold decrease in turnover number for dTDP-glucose compared to wild-type value, 2fold increase in KM-value for dTDP-glucose compared to wild-type value. Less than 0.5% of the mutant enzyme contains NADH during steady-state turnover by adding a large excess of dTDPglucose, compared to 45% of the wild-type enzyme Escherichia coli
E136Q 67fold decrease in turnover number for dTDP-glucose compared to wild-type value, 1.5fold increase in KM-value for dTDP-glucose compared to wild-type value. 3% of the mutant enzyme contains NADH during steady-state turnover by adding a large excess of dTDPglucose, compared to 45% of the wild-type enzyme Escherichia coli
E198Q 258fold decrease in turnover number for dTDP-glucose compared to wild-type value, 1.8fold increase in KM-value for dTDP-glucose compared to wild-type value. Less than 0.5% of the mutant enzyme contains NADH during steady-state turnover by adding a large excess of dTDPglucose, compared to 45% of the wild-type enzyme Escherichia coli
H232A 57.6fold decrease in turnover number for dTDP-glucose compared to wild-type value, 1.8fold decrease in KM-value for dTDP-glucose compared to wild-type value Escherichia coli
H232N 6.8fold decrease in turnover number for dTDP-glucose compared to wild-type value, 1.2fold decrease in KM-value for dTDP-glucose compared to wild-type value. 3% of the mutant enzyme contains NADH during steady-state turnover by adding a large excess of dTDPglucose, compared to 45% of the wild-type enzyme Escherichia coli
H232Q 114fold decrease in turnover number for dTDP-glucose compared to wild-type value, 1.3fold increase in KM-value for dTDP-glucose compared to wild-type value. Less than 0.5% of the mutant enzyme contains NADH during steady-state turnover by adding a large excess of dTDPglucose, compared to 45% of the wild-type enzyme Escherichia coli
K199M 115fold decrease in turnover number for dTDP-glucose compared to wild-type value, 1.3fold increase in KM-value for dTDP-glucose compared to wild-type value. 2% of the mutant enzyme contains NADH during steady-state turnover by adding a large excess of dTDPglucose, compared to 45% of the wild-type enzyme Escherichia coli
K199R 288fold decrease in turnover number for dTDP-glucose compared to wild-type value, 2.2fold increase in KM-value for dTDP-glucose compared to wild-type value. 2% of the mutant enzyme contains NADH during steady-state turnover by adding a large excess of dTDPglucose, compared to 45% of the wild-type enzyme Escherichia coli
N190A 551fold decrease in turnover number for dTDP-glucose compared to wild-type value, 0.92fold increase in KM-value for dTDP-glucose compared to wild-type value. Less than 0.5% of the mutant enzyme contains NADH during steady-state turnover by adding a large excess of dTDPglucose, compared to 45% of the wild-type enzyme Escherichia coli
N190D 441fold decrease in turnover number for dTDP-glucose compared to wild-type value, fold 4.2increase in KM-value for dTDP-glucose compared to wild-type value. Less than 0.5% of the mutant enzyme contains NADH during steady-state turnover by adding a large excess of dTDPglucose, compared to 45% of the wild-type enzyme Escherichia coli
N190H 217fold decrease in turnover number for dTDP-glucose compared to wild-type value, 1.2fold increase in KM-value for dTDP-glucose compared to wild-type value Escherichia coli
Y301F 117fold decrease in turnover number for dTDP-glucose compared to wild-type value, 5.2fold increase in KM-value for dTDP-glucose compared to wild-type value. 39% of the mutant enzyme contains NADH during steady-state turnover by adding a large excess of dTDPglucose, compared to 45% of the wild-type enzyme Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
dTDP-galactose
-
 Escherichia coli
dTDP-xylose
-
 Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.001
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme C187A Escherichia coli
0.0015
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme C187S Escherichia coli
0.0033
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme H232A Escherichia coli
0.005
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme H232N Escherichia coli
0.0055
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme N190A Escherichia coli
0.006
-
 dTDP-glucose pH 7.5, 37°C, wild-type enzyme Escherichia coli
0.007
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme D135A Escherichia coli
0.0073
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme N190H Escherichia coli
0.0079
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme D135N Escherichia coli
0.008
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme H232Q Escherichia coli
0.008
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme K199M Escherichia coli
0.009
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme E136Q Escherichia coli
0.011
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme E198Q Escherichia coli
0.012
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme E136A Escherichia coli
0.013
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme K199R Escherichia coli
0.019
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme D135N/E136Q Escherichia coli
0.025
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme N190D Escherichia coli
0.031
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme Y301F Escherichia coli
2.2
-
 dTDP-glucose pH 7.5, 37°C, wild-type enzyme Escherichia coli
30
-
 UDP-glucose pH 7.5, 37°C, wild-type enzyme Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli P27830
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
dTDP-glucose
-
 Escherichia coli dTDP-4-dehydro-6-deoxy-D-glucose + H2O
-
 ?
UDP-D-glucose
-
 Escherichia coli UDP-4-dehydro-6-deoxy-D-glucose + H2O
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0089
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme N190A Escherichia coli
0.011
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme N190D Escherichia coli
0.017
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme E136A Escherichia coli
0.017
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme K199R Escherichia coli
0.019
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme E198Q Escherichia coli
0.022
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme D135A Escherichia coli
0.0226
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme N190H Escherichia coli
0.024
-
 UDP-glucose pH 7.5, 37°C, wild-type enzyme Escherichia coli
0.024
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme D135N/E136Q Escherichia coli
0.0395
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme D135N Escherichia coli
0.042
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme Y301F Escherichia coli
0.0425
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme K199M Escherichia coli
0.043
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme H232Q Escherichia coli
0.073
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme E136Q Escherichia coli
0.085
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme H232A Escherichia coli
0.52
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme C187A Escherichia coli
0.72
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme H232N Escherichia coli
1.2
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme C187S Escherichia coli
1.9
-
 dTDP-glucose pH 7.5, 18°C, wild-type enzyme Escherichia coli
4.9
-
 dTDP-glucose pH 7.5, 37°C, wild-type enzyme Escherichia coli
6.08
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme C187A Escherichia coli
6.08
-
 dTDP-glucose pH 7.5, 37°C, mutant enzyme H232N Escherichia coli

Cofactor

Cofactor Comment Organism Structure
NADH 45% of the wild-type enzyme contains NADH during steady-state turnover for each variant by adding a large excess of dTDPglucose, 9% of mutant enzyme D135N, 30% of mutant enzyme D135A, 3% of mutant enzyme E136Q and H232N, 2% of mutant enzyme K199M and K199R, 39% of mutant enzyme Y301F, 8% of mutant enzyme C187A and less than 0.5% of mutant enzymes D135N/E136Q, E198Q, N190D, N190A and H232Q Escherichia coli

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0112
-
 dTDP-xylose pH 7.5, 37°C, wild-type enzyme Escherichia coli
0.097
-
 dTDP-galactose pH 7.5, 37°C, wild-type enzyme Escherichia coli