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

BRENDA support

Literature summary for 5.3.3.1 extracted from

  • Choi, G.; Ha, N.C.; Kim, S.W.; Kim, D.H.; Park, S.; Oh, B.H.; Choi, K.Y.
    Asp-99 donates a hydrogen bond not to Tyr-14 but to the steroid directly in the catalytic mechanism of DELTA 5-3-ketosteroid isomerase from Pseudomonas putida biotype B (2000), Biochemistry, 39, 903-909.
    View publication on PubMed

Crystallization (Commentary)

Crystallization (Comment) Organism
mutant enzyme D99E/D38N complexed with equilenin, an intermediate analogue, crystals of the complexes are grown from 1.1 M ammonium acetate and 0.1 M sodium acetate, pH 4.6, by the hanging drop vapor diffusion method at 22°C. The resulting crystals have C2 space group symmetry with unit cell dimensions of a = 89.04 A, b = 72.42 A, c = 51.24 A and beta = 90.9° Pseudomonas putida

Protein Variants

Protein Variants Comment Organism
D99E turnover-number for 5-androstene-3,17-dione is 68.1% of the turnover number of the wild-type enzyme, the Km-value is 1.22fold higher than the Km-value of the wild-type enzyme Pseudomonas putida
D99L turnover-number for 5-androstene-3,17-dione is 125fold lower than the turnover number of the wild-type enzyme, the Km-value is 1.95fold lower than the Km-value of the wild-type enzyme Pseudomonas putida
Y14F turnover-number for 5-androstene-3,17-dione is less than 1% of the turnover number of the wild-type enzyme, the Km-value is 1.87fold lower than the Km-value of the wild-type enzyme Pseudomonas putida
Y14F/D99E turnover-number for 5-androstene-3,17-dione is less than 1% of the turnover number of the wild-type enzyme, the Km-value is 1.05fold lower than the Km-value of the wild-type enzyme Pseudomonas putida
Y14F/D99L turnover-number for 5-androstene-3,17-dione is less than 1% of the turnover number of the wild-type enzyme, the Km-value is 1.95fold higher than the Km-value of the wild-type enzyme Pseudomonas putida
Y14F/D99N turnover-number for 5-androstene-3,17-dione is less than 1% of the turnover number of the wild-type enzyme, the Km-value is 1.06fold lower than the Km-value of the wild-type enzyme Pseudomonas putida

Inhibitors

Inhibitors Comment Organism Structure
19-nortestosterone
-
Pseudomonas putida
equilenin
-
Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0269
-
5-androstene-3,17-dione pH 7.0, 25°C, mutant enzyme Y14F Pseudomonas putida
0.0475
-
5-androstene-3,17-dione pH 7.0, 25°C, mutant enzyme Y14F/D99N Pseudomonas putida
0.0481
-
5-androstene-3,17-dione pH 7.0, 25°C, mutant enzyme Y14F/D99E Pseudomonas putida
0.0503
-
5-androstene-3,17-dione pH 7.0, 25°C, wild-type enzyme Pseudomonas putida
0.0616
-
5-androstene-3,17-dione pH 7.0, 25°C, mutant enzyme D99E Pseudomonas putida
0.0982
-
5-androstene-3,17-dione pH 7.0, 25°C, mutant enzyme Y14F/D99L Pseudomonas putida
0.258
-
5-androstene-3,17-dione pH 7.0, 25°C, mutant enzyme D99L Pseudomonas putida

Organism

Organism UniProt Comment Textmining
Pseudomonas putida
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5-Androstene-3,17-dione
-
Pseudomonas putida 4-Androstene-3,17-dione
-
?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.67
-
5-androstene-3,17-dione pH 7.0, 25°C, mutant enzyme Y14F/D99L Pseudomonas putida
0.7 1 5-androstene-3,17-dione pH 7.0, 25°C, mutant enzyme Y14F/D99N Pseudomonas putida
0.71
-
5-androstene-3,17-dione pH 7.0, 25°C, mutant enzyme Y14F/D99N Pseudomonas putida
3
-
5-androstene-3,17-dione pH 7.0, 25°C, mutant enzyme Y14F/D99E Pseudomonas putida
6.08
-
5-androstene-3,17-dione pH 7.0, 25°C, mutant enzyme Y14F/D99L Pseudomonas putida
11.5
-
5-androstene-3,17-dione pH 7.0, 25°C, mutant enzyme Y14F Pseudomonas putida
220
-
5-androstene-3,17-dione pH 7.0, 25°C, mutant enzyme D99L Pseudomonas putida
19000
-
5-androstene-3,17-dione pH 7.0, 25°C, mutant enzyme D99E Pseudomonas putida
27900
-
5-androstene-3,17-dione pH 7.0, 25°C, wild-type enzyme Pseudomonas putida

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
additional information
-
additional information Ki-values for equilenin and 19-nortestosterone, wild-type enzyme and mutant enzyme D99E Pseudomonas putida