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Literature summary for 5.3.3.1 extracted from

  • Jang, D.S.; Choi, G.; Cha, H.J.; Shin, S.; Hong, B.H.; Lee, H.J.; Lee, H.C.; Choi, K.Y.
    Contribution of a low-barrier hydrogen bond to catalysis is not significant in ketosteroid isomerase (2015), Mol. Cells, 38, 409-415 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 5.3.3.1

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Pseudomonas putida

Crystallization (Commentary)

Crystallization (Comment) Organism
mutant enzyme Y30F/Y55F/Y115F/D38N KSI complexed with equilenin, hanging drop vapor diffusion method, using 0.1 M sodium acetate, pH 4.5, 0.6 M ammonium acetate, and 30% PEG 4000 Pseudomonas putida

Protein Variants

Protein Variants Comment Organism
Y115F the mutant shows about 68% activity compared to the wild type enzyme Pseudomonas putida
Y14F the mutant shows almost no activity compared to the wild type enzyme Pseudomonas putida
Y30F the mutant shows about 84% activity compared to the wild type enzyme Pseudomonas putida
Y30F/Y55F the mutant shows about 50% activity compared to the wild type enzyme Pseudomonas putida
Y30F/Y55F/Y115F the mutant shows about 57% activity compared to the wild type enzyme Pseudomonas putida
Y55F the mutant shows about 17% activity compared to the wild type enzyme Pseudomonas putida

Inhibitors

Inhibitors Comment Organism Structure
androstenolone
-
 Pseudomonas putida
equilenin
-
 Pseudomonas putida
estrone
-
 Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0171
-
 5-androstene-3,17-dione mutant enzyme Y14F, at pH 7.0, temperature not specified in the publication Pseudomonas putida
0.023
-
 5-androstene-3,17-dione mutant enzyme Y55F, at pH 7.0, temperature not specified in the publication Pseudomonas putida
0.0417
-
 5-androstene-3,17-dione mutant enzyme Y115F, at pH 7.0, temperature not specified in the publication Pseudomonas putida
0.0499
-
 5-androstene-3,17-dione wild type enzyme, at pH 7.0, temperature not specified in the publication Pseudomonas putida
0.0502
-
 5-androstene-3,17-dione mutant enzyme Y30F/Y55F, at pH 7.0, temperature not specified in the publication Pseudomonas putida
0.0552
-
 5-androstene-3,17-dione mutant enzyme Y30F, at pH 7.0, temperature not specified in the publication Pseudomonas putida
0.0773
-
 5-androstene-3,17-dione mutant enzyme Y30F/Y55F/Y115F, at pH 7.0, temperature not specified in the publication Pseudomonas putida

Organism

Organism UniProt Comment Textmining
Pseudomonas putida P07445
-
-

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
-
 ?

Synonyms

Synonyms Comment Organism
DELTA5-3-ketosteroid isomerase
-
 Pseudomonas putida
KSI
-
 Pseudomonas putida

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
13.3
-
 5-androstene-3,17-dione mutant enzyme Y14F, at pH 7.0, temperature not specified in the publication Pseudomonas putida
3510
-
 5-androstene-3,17-dione mutant enzyme Y55F, at pH 7.0, temperature not specified in the publication Pseudomonas putida
10700
-
 5-androstene-3,17-dione mutant enzyme Y30F/Y55F, at pH 7.0, temperature not specified in the publication Pseudomonas putida
12200
-
 5-androstene-3,17-dione mutant enzyme Y30F/Y55F/Y115F, at pH 7.0, temperature not specified in the publication Pseudomonas putida
14500
-
 5-androstene-3,17-dione mutant enzyme Y115F, at pH 7.0, temperature not specified in the publication Pseudomonas putida
17800
-
 5-androstene-3,17-dione mutant enzyme Y30F, at pH 7.0, temperature not specified in the publication Pseudomonas putida
21000
-
 5-androstene-3,17-dione wild type enzyme, at pH 7.0, temperature not specified in the publication Pseudomonas putida

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
780
-
 5-androstene-3,17-dione mutant enzyme Y14F, at pH 7.0, temperature not specified in the publication Pseudomonas putida
150000
-
 5-androstene-3,17-dione mutant enzyme Y55F, at pH 7.0, temperature not specified in the publication Pseudomonas putida
160000
-
 5-androstene-3,17-dione mutant enzyme Y30F/Y55F/Y115F, at pH 7.0, temperature not specified in the publication Pseudomonas putida
210000
-
 5-androstene-3,17-dione mutant enzyme Y30F/Y55F, at pH 7.0, temperature not specified in the publication Pseudomonas putida
330000
-
 5-androstene-3,17-dione mutant enzyme Y30F, at pH 7.0, temperature not specified in the publication Pseudomonas putida
350000
-
 5-androstene-3,17-dione mutant enzyme Y115F, at pH 7.0, temperature not specified in the publication Pseudomonas putida
430000
-
 5-androstene-3,17-dione wild type enzyme, at pH 7.0, temperature not specified in the publication Pseudomonas putida