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Literature summary for 1.3.1.34 extracted from

  • Tu, X.; Hubbard, P.A.; Kim, J.J.; Schulz, H.
    Two distinct proton donors at the active site of Escherichia coli 2,4-dienoyl-CoA reductase are responsible for the formation of different products (2008), Biochemistry, 47, 1167-1175.
    View publication on PubMed
Search for more literature for 1.3.1.34

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21 cells Escherichia coli

Protein Variants

Protein Variants Comment Organism
C334A/C337A the mutation exhibits 0.02% of wild type activity Escherichia coli
C337A the mutation results in the loss of most of the iron and acid-labile sulfur and exhibits 0.09% of wild type activity Escherichia coli
E164A mutant exhibits 1.5% of the wild type activity Escherichia coli
H252A the mutation exhibits 0.09% of wild type activity Escherichia coli
H252F mutant is inactive Escherichia coli
Y166F mutant exhibits 27% of the wild type activity, however, the product of the reduction is 3-enoyl-CoA instead of 2-enoyl-CoA Escherichia coli
Y166F/E164Q double mutant is inactive Escherichia coli
Y166F/H252A the double mutation exhibits 4.4% of the wild type activity Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00044
-
 NADPH mutant enzyme Y166F, in 50 mM potassium phosphate buffer (pH 7.4) Escherichia coli
0.0008
-
 NADPH wild type enzyme, in 50 mM potassium phosphate buffer (pH 7.4) Escherichia coli
0.00141
-
 NADPH mutant enzyme C337A, in 50 mM potassium phosphate buffer (pH 7.4) Escherichia coli
0.0024
-
 NADPH mutant enzyme H252A/Y166F, in 50 mM potassium phosphate buffer (pH 7.4) Escherichia coli
0.00534
-
 NADPH mutant enzyme E164A, in 50 mM potassium phosphate buffer (pH 7.4) Escherichia coli
0.00548
-
 NADPH mutant enzyme H252A, in 50 mM potassium phosphate buffer (pH 7.4) Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Iron the wild type enzyme contains 3.9 mol iron per protein Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli P42593
-
-

Purification (Commentary)

Purification (Comment) Organism
NTA-Ni2+ agarose column chromatography Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0015
-
 mutant enzyme C334A/C337A, with 2-trans,4-trans-decadienoyl-CoA as substrate Escherichia coli
0.0067
-
 mutant enzyme C337A, with 2-trans,4-trans-decadienoyl-CoA as substrate Escherichia coli
0.007
-
 mutant enzyme H252A, with 2-trans,4-trans-decadienoyl-CoA as substrate Escherichia coli
0.095
-
 mutant enzyme E164A, with 2-trans,4-trans-decadienoyl-CoA as substrate Escherichia coli
0.33
-
 mutant enzyme H252A/Y166F, with 2-trans,4-trans-decadienoyl-CoA as substrate Escherichia coli
2.02
-
 mutant enzyme Y166F, with 2-trans,4-trans-decadienoyl-CoA as substrate Escherichia coli
7.54
-
 wild type enzyme, with 2-trans,4-trans-decadienoyl-CoA as substrate Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-trans,4-trans-decadienoyl-CoA + NADPH
-
 Escherichia coli 2-trans-decenoyl-CoA + NADP+ 2-trans-decenoyl-CoA is the normal product of the wild type enzyme in bacteria ?
2-trans,4-trans-decadienoyl-CoA + NADPH
-
 Escherichia coli 3-trans-decenoyl-CoA + NADP+ 3-trans-decenoyl-CoA is the product of the mutant enzymes Y166F, H252A, and H252A/Y166F ?

Synonyms

Synonyms Comment Organism
DCR
-
 Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.008
-
 NADPH mutant enzyme C337A, in 50 mM potassium phosphate buffer (pH 7.4) Escherichia coli
0.011
-
 NADPH mutant enzyme H252A, in 50 mM potassium phosphate buffer (pH 7.4) Escherichia coli
0.15
-
 NADPH mutant enzyme E164A, in 50 mM potassium phosphate buffer (pH 7.4) Escherichia coli
0.44
-
 NADPH mutant enzyme H252A/Y166F, in 50 mM potassium phosphate buffer (pH 7.4) Escherichia coli
2.5
-
 NADPH mutant enzyme Y166F, in 50 mM potassium phosphate buffer (pH 7.4) Escherichia coli
9.5
-
 NADPH wild type enzyme, in 50 mM potassium phosphate buffer (pH 7.4) Escherichia coli

Cofactor

Cofactor Comment Organism Structure
4Fe-4S-center
-
 Escherichia coli
NADPH dependent Escherichia coli