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Literature summary for 5.3.3.1 extracted from

  • Kim, D.H.; Nam, G.H.; Jang, D.S.; Choi, G.; Joo, S.; Kim, J.S.; Oh, B.H.; Choi, K.Y.
    Roles of active site aromatic residues in catalysis by ketosteroid isomerase from Pseudomonas putida biotype B (1999), Biochemistry, 38, 13810-13819.
    View publication on PubMed
Search for more literature for 5.3.3.1

Crystallization (Commentary)

Crystallization (Comment) Organism
crystal structure of mutant enzyme F82A is determined to 2.1 A resolution. Crystals are grown in a solution containing 1.0 M sodium acetate and 0.1 M ammonioum acetate, pH 4.6, by the hanging drop method of vapor diffusion at 22 C. The crystals belong to the space group c2221 with unit cell dimensions of a = 36.24 A, b = 96.13 A and c = 74.30 A Pseudomonas putida

Protein Variants

Protein Variants Comment Organism
F54A turnover number for 5-androstene-3,17-dione is 17.1% of the turnover number for the wild-type enzyme, turnover number for 5(10)-estrene-3,17-dione is 15.3% of the turnover number of the wild-type enzyme,the KM-value for 5-androstene-3,17-dione is 116% of the KM-value of the wild-type enzyme, the KM-value for 5(10)-estrene-3,17-dione is 233% of the KM-value of the wild-type enzyme Pseudomonas putida
F54L turnover number for 5-androstene-3,17-dione is 41.9% of the turnover number of the wild-type enzyme, turnover number for 5(10)-estrene-3,17-dione is 38.7% of the turnover number of the wild-type enzyme, the KM-value for 5-androstene-3,17-dione is 154% of the KM-value of the wild-type enzyme, the KM-value for 5(10)-estrene-3,17-dione is 89.3% of the KM-value of the wild-type enzyme Pseudomonas putida
F82A turnover number for 5-androstene-3,17-dione is 2.5% of the turnover number of the wild-type enzyme, turnover number for 5(10)-estrene-3,17-dione is 2.3% of the turnover number of the wild-type enzyme,the KM-value for 5-androstene-3,17-dione is 29.9% of the KM-value of the wild-type enzyme, the KM-value for 5(10)-estrene-3,17-dione is 29.8% of the KM-value of the wild-type enzyme Pseudomonas putida
F82L turnover number for 5-androstene-3,17-dione is 12% of the turnover number of the wild-type enzyme, turnover number for 5(10)-estrene-3,17-dione is 67.6% of the turnover number of the wild-type enzyme, the KM-value for 5-androstene-3,17-dione is 108% of the KM-value of the wild-type enzyme, the KM-value for 5(10)-estrene-3,17-dione is 130% of the KM-value of the wild-type enzyme Pseudomonas putida
W116A turnover number for 5-androstene-3,17-dione is 1.5% of the turnover number of the wild-type enzyme, turnover number for 5(10)-estrene-3,17-dione is below 12%% of the turnover number of the wild-type enzyme, the KM-value for 5-androstene-3,17-dione is 348% of the KM-value of the wild-type enzyme Pseudomonas putida
W116F turnover number for 5-androstene-3,17-dione is 22.4% of the turnover number of the wild-type enzyme, the KM-value for 5(10)-estrene-3,17-dione is 299% of the KM-value of the wild-type enzyme Pseudomonas putida
W116Y turnover number for 5-androstene-3,17-dione is 21% of the turnover number of the wild-type enzyme Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0149
-
 5-androstene-3,17-dione 25°C, pH 7.0, mutant enzyme F82A Pseudomonas putida
0.0328
-
 5(10)-estrene-3,17-dione 25°C, pH 7.0, mutant enzyme F82A Pseudomonas putida
0.0499
-
 5-androstene-3,17-dione 25°C, pH 7.0, wild-type enzyme Pseudomonas putida
0.0539
-
 5-androstene-3,17-dione 25°C, pH 7.0, mutant enzyme F82L Pseudomonas putida
0.0579
-
 5-androstene-3,17-dione 25°C, pH 7.0, mutant enzyme F54A Pseudomonas putida
0.0651
-
 5(10)-estrene-3,17-dione 25°C, pH 7.0, mutant enzyme F82L Pseudomonas putida
0.077
-
 5-androstene-3,17-dione 25°C, pH 7.0, mutant enzyme F54L Pseudomonas putida
0.0982
-
 5(10)-estrene-3,17-dione 25°C, pH 7.0, mutant enzyme F54L Pseudomonas putida
0.11
-
 5(10)-estrene-3,17-dione 25°C, pH 7.0, wild-type enzyme Pseudomonas putida
0.149
-
 5-androstene-3,17-dione 25°C, pH 7.0, mutant enzyme W116F Pseudomonas putida
0.174
-
 5-androstene-3,17-dione 25°C, pH 7.0, mutant enzyme W116A Pseudomonas putida
0.256
-
 5(10)-estrene-3,17-dione 25°C, pH 7.0, mutant enzyme F54A Pseudomonas putida

Organism

Organism UniProt Comment Textmining
Pseudomonas putida
-
 biotype B
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5(10)-estrene-3,17-dione
-
 Pseudomonas putida estr-4-en-3,17-dione
-
 ?
5-Androstene-3,17-dione
-
 Pseudomonas putida 4-Androstene-3,17-dione
-
 ?
additional information the aromatic residues Phe54, Phe82 and Trp116 in the hydrophobic substrate-binding pocket of the enzyme contribute differentially to steroid species including substrate, intermediate and product Pseudomonas putida ?
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.25
-
 5(10)-estrene-3,17-dione 25°C, pH 7.0, mutant enzyme F82A Pseudomonas putida
1.7
-
 5(10)-estrene-3,17-dione 25°C, pH 7.0, mutant enzyme F54A Pseudomonas putida
4.3
-
 5(10)-estrene-3,17-dione 25°C, pH 7.0, mutant enzyme F54L Pseudomonas putida
7.5
-
 5(10)-estrene-3,17-dione 25°C, pH 7.0, mutant enzyme F82L Pseudomonas putida
11.1
-
 5(10)-estrene-3,17-dione 25°C, pH 7.0, wild-type enzyme Pseudomonas putida
311
-
 5-androstene-3,17-dione 25°C, pH 7.0, mutant enzyme W116A Pseudomonas putida
539
-
 5-androstene-3,17-dione 25°C, pH 7.0, mutant enzyme F82A Pseudomonas putida
2540
-
 5-androstene-3,17-dione 25°C, pH 7.0, mutant enzyme F82L Pseudomonas putida
3630
-
 5-androstene-3,17-dione 25°C, pH 7.0, mutant enzyme F54A Pseudomonas putida
4420
-
 5-androstene-3,17-dione 25°C, pH 7.0, mutant enzyme W116Y Pseudomonas putida
4760
-
 5-androstene-3,17-dione 25°C, pH 7.0, mutant enzyme W116F Pseudomonas putida
8900
-
 5-androstene-3,17-dione 25°C, pH 7.0, mutant enzyme F54L Pseudomonas putida
21200
-
 5-androstene-3,17-dione 25°C, pH 7.0, wild-type enzyme Pseudomonas putida
21230
-
 5-androstene-3,17-dione 25°C, pH 7.0, wild-type enzyme Pseudomonas putida