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Literature summary for 1.1.3.6 extracted from

  • Nishiya, Y.; Harada, N.; Teshima, S-i.; Yamahita, M.; Fuji, I.; Hirayama, N.; Murooka, Y.
    Improvement of thermal stability of Streptomyces cholesterol oxidase by random mutagenesis and a structural interpretation (1997), Protein Eng., 10, 231-235.
    View publication on PubMed
Search for more literature for 1.1.3.6

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Streptomyces sp.

Protein Variants

Protein Variants Comment Organism
R135H random mutagenesis Streptomyces sp.
S103T random mutagenesis Streptomyces sp.
V121A random mutagenesis Streptomyces sp.
V145E random mutagenesis Streptomyces sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.011
-
 cholesterol V145E mutant Streptomyces sp.
0.013
-
 cholesterol wild-type Streptomyces sp.
0.013
-
 cholesterol S103T mutant Streptomyces sp.
0.03
-
 cholesterol R135H mutant Streptomyces sp.

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
cholesterol + O2 Streptomyces sp.
-
 cholest-5-en-3-one + H2O2
-
 ?

Organism

Organism UniProt Comment Textmining
Streptomyces sp.
-
-
-

Reaction

Reaction Comment Organism Reaction ID
cholesterol + O2 = cholest-5-en-3-one + H2O2 bifunctional enzyme, catalyzes both the oxidation of DELTA5-ene-3beta hydroxysteroids with a trans A-B ring junction to the corresponding DELTA5-3-ketosteroid with the reduction of oxygen to hydrogen peroxide, and the isomerization to the DELTA4-3-ketosteroid Streptomyces sp.
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
26.7
-
 S103T/V145A/V121A/R135H quadruple mutant Streptomyces sp.
39.9
-
 S103T/V145A double mutant Streptomyces sp.
47.8
-
 S103T/V145A/V121A triple mutant Streptomyces sp.
63.5
-
 V145E mutant, 37°C, pH 7.0 Streptomyces sp.
67.7
-
 wild type Streptomyces sp.
68.9
-
 R135H mutant, 37°C, pH 7.0 Streptomyces sp.
72
-
 V121A mutant, 37°C, pH 7.0 Streptomyces sp.
86.6
-
 S103T mutant, 37°C, pH 7.0 Streptomyces sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
cholesterol + O2
-
 Streptomyces sp. cholest-4-en-3-one + H2O2
-
 ?
cholesterol + O2
-
 Streptomyces sp. cholest-5-en-3-one + H2O2
-
 ?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
60
-
 wild-type and S103T, V121A, R135H and V145E mutants Streptomyces sp.

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
60
-
 half-lives of S103T, V121A, R135H, V145E mutants and wild-type: 11.3, 12.2, 11.0, 24.1 and 7.8 min respectively, half-lives of S103T/V145A double mutants and S103T/V145A/V121A triple mutants: 32.4 and 52.2 min respectively Streptomyces sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
 wild-type, S103T, V121A and R135H mutants Streptomyces sp.
7.5
-
 V145E mutant Streptomyces sp.

pH Range

pH Minimum pH Maximum Comment Organism
6 8 wild-type, S103T, V121A, R135H and V145E mutants Streptomyces sp.