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Literature summary for 5.3.3.1 extracted from

  • Henot, F.; Pollack, R.M.
    Catalytic activity of the D38A mutant of 3-oxo-DELTA 5-steroid isomerase: recruitment of aspartate-99 as the base (2000), Biochemistry, 39, 3351-3359.
    View publication on PubMed
Search for more literature for 5.3.3.1

Cloned(Commentary)

Cloned (Comment) Organism
D38A, D38A/Y14F, D38A/Y55F and D38A/D99N proteins are expressed in Epicurian Coli XL1-Blue cells Comamonas testosteroni

Protein Variants

Protein Variants Comment Organism
D38A the catalytic turnover number is 140fold less than that for the wild-type Comamonas testosteroni
D38A/D99N no detectable activity Comamonas testosteroni
D38A/Y14F no detectable activity Comamonas testosteroni
D38A/Y55F no detectable activity Comamonas testosteroni

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.067
-
 5-androstene-3,17-dione 25°C, pH 6.5, solvent D2O, mutant enzyme D38A Comamonas testosteroni
0.073
-
 5-androstene-3,17-dione 25°C, pH 8.5, solvent D2O, mutant enzyme D38A Comamonas testosteroni
0.085
-
 5-androstene-3,17-dione 25°C, pH 7.5, solvent D2O, mutant enzyme D38A Comamonas testosteroni
0.094
-
 5-androstene-3,17-dione 25°C, pH 8.5, solvent H2O, mutant enzyme D38A Comamonas testosteroni
0.096
-
 5-androstene-3,17-dione 25°C, pH 7.5, solvent H2O, mutant enzyme D38A Comamonas testosteroni
0.099
-
 5-androstene-3,17-dione 25°C, pH 6.5, solvent H2O, mutant enzyme D38A Comamonas testosteroni

Organism

Organism UniProt Comment Textmining
Comamonas testosteroni
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5-Androstene-3,17-dione residue Asp38 acts as a base to abstract a proton from C-4 of the substrate to form an intermediate dienolate, which is then reprotonated on C-6 Comamonas testosteroni 4-Androstene-3,17-dione
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
230
-
 5-androstene-3,17-dione 25°C, pH 6.5, solvent D2O, mutant enzyme D38A Comamonas testosteroni
260
-
 5-androstene-3,17-dione 25°C, pH 8.5, solvent D2O, mutant enzyme D38A Comamonas testosteroni
279
-
 5-androstene-3,17-dione 25°C, pH 6.5, solvent H2O, mutant enzyme D38A Comamonas testosteroni
340
-
 5-androstene-3,17-dione 25°C, pH 6.5, solvent H2O, mutant enzyme D38A Comamonas testosteroni
412
-
 5-androstene-3,17-dione 25°C, pH 7.5 or 8.5, solvent H2O, mutant enzyme D38A Comamonas testosteroni