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Literature summary for 1.1.3.6 extracted from

  • Kasabe, P.J.; Mali, G.T.; Dandge, P.B.
    Assessment of alkaline cholesterol oxidase purified from Rhodococcus sp. PKPD-CL for its halo tolerance, detergent and organic solvent stability (2015), Protein Expr. Purif., 116, 30-41 .
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
Benzene 9% activation at 0.5% w/v Rhodococcus sp.
chloroform 7% activation at 0.5% w/v Rhodococcus sp.
ethanol 10% activation at 0.5% w/v Rhodococcus sp.
Isopropanol 15% activation at 0.5% w/v Rhodococcus sp.
NaN3 activates slightly Rhodococcus sp.
Tween-80 18% activation at 0.5% w/v Rhodococcus sp.

General Stability

General Stability Organism
purified enzyme, 80% residual activity with 20% NaCl (3.42 M) and 83% relative activity with 12% NaCl (2.05 M) concentration. The enzyme is stable in presence of chemical reagents (NaN3, EDTA), detergents (Tween-80, Tween-20, Triton X-100, sodium cholate) Rhodococcus sp.

Inhibitors

Inhibitors Comment Organism Structure
4-chloromercuribenzoate 85% inhibition at 0.1 mM Rhodococcus sp.
acetone 85% inhibition at a ratio of 1:2 with enzyme solution Rhodococcus sp.
Ag+ 75% inhibition at 0.1 mM Rhodococcus sp.
Ba2+ 91% inhibition at 0.1 mM Rhodococcus sp.
butanol 6% inhibition at a ratio of 1:2 with enzyme solution Rhodococcus sp.
Ca2+
-
Rhodococcus sp.
Cu2+ 69% inhibition at 0.1 mM Rhodococcus sp.
DMSO 11% inhibition at a ratio of 1:2 with enzyme solution Rhodococcus sp.
Fe2+ 10% inhibition at 0.1 mM Rhodococcus sp.
Fe3+ 88% inhibition at 0.1 mM Rhodococcus sp.
Hg2+ complete inhibition at 0.1 mM Rhodococcus sp.
Li+ 8% inhibition at 0.1 mM Rhodococcus sp.
Mg2+
-
Rhodococcus sp.
additional information the enzyme activity is unaffected or poorly affected by Tween-20, Triton X-100, sodium cholate, methanol, toluene, and ethyl acetate Rhodococcus sp.
NaCl 80% residual activity with 20% NaCl (3.42 M) and 83% relative activity with 12% NaCl (2.05 M) concentration Rhodococcus sp.
SDS 88% inhibition at 0.5% w/v Rhodococcus sp.
Zn2+ 67% inhibition at 0.1 mM Rhodococcus sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.11
-
cholesterol pH 8.0, 37°C Rhodococcus sp.

Localization

Localization Comment Organism GeneOntology No. Textmining
extracellular
-
Rhodococcus sp.
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
additional information the enzyme activity is unaffected or poorly affected by Ca2+, Mg2+, Mn2+, and EDTA Rhodococcus sp.

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
cholesterol + O2 Rhodococcus sp.
-
cholest-4-en-3-one + H2O2
-
?
cholesterol + O2 Rhodococcus sp. PKPD-CL
-
cholest-4-en-3-one + H2O2
-
?

Organic Solvent Stability

Organic Solvent Comment Organism
benzene stable in presence of Rhodococcus sp.
Butanol stable in presence of Rhodococcus sp.
chloroform stable in presence of Rhodococcus sp.
dimethyl sulfoxide stable in presence of Rhodococcus sp.
Ethanol stable in presence of Rhodococcus sp.
Ethyl acetate stable in presence of Rhodococcus sp.
isopropanol stable in presence of Rhodococcus sp.
Methanol stable in presence of Rhodococcus sp.
toluene stable in presence of Rhodococcus sp.

Organism

Organism UniProt Comment Textmining
Rhodococcus sp.
-
isolated from brackish water of the Chilika Lake at Odisha state of India
-
Rhodococcus sp. PKPD-CL
-
isolated from brackish water of the Chilika Lake at Odisha state of India
-

Purification (Commentary)

Purification (Comment) Organism
extracellular enzyme 59fold from strain PKPD-CL by ammonium sulfate fractionation, anion exchange chromatography, ultrafiltration, and gel filtration Rhodococcus sp.

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
35.64
-
purified enzyme, pH 8.0, 37°C Rhodococcus sp.

Storage Stability

Storage Stability Organism
30°C, purified enzyme, completely stable for 60 days, 24% reduction after 70 days, 60% reduction after 100 days Rhodococcus sp.
4°C, purified enzyme, completely stable for 60 days, 24% reduction after 70 days, 60% reduction after 100 days Rhodococcus sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
beta-sitosterol + O2 85% activity compared to cholesterol Rhodococcus sp. stigmast-5-en-3-one + H2O2
-
?
beta-sitosterol + O2 85% activity compared to cholesterol Rhodococcus sp. PKPD-CL stigmast-5-en-3-one + H2O2
-
?
beta-stigmasterol + O2 83% activity compared to cholesterol Rhodococcus sp. stigmasta-5,22-dien-3-one + H2O2
-
?
beta-stigmasterol + O2 83% activity compared to cholesterol Rhodococcus sp. PKPD-CL stigmasta-5,22-dien-3-one + H2O2
-
?
cholestenol + O2 43% activity compared to cholesterol Rhodococcus sp. 5alpha-cholest-8-en-3-one + H2O2
-
?
cholesterol + O2
-
Rhodococcus sp. cholest-4-en-3-one + H2O2
-
?
cholesterol + O2 best substrate Rhodococcus sp. cholest-4-en-3-one + H2O2
-
?
cholesterol + O2
-
Rhodococcus sp. PKPD-CL cholest-4-en-3-one + H2O2
-
?
cholesterol + O2 best substrate Rhodococcus sp. PKPD-CL cholest-4-en-3-one + H2O2
-
?
ergosterol + O2 22% activity compared to cholesterol Rhodococcus sp. ? + H2O2
-
?
additional information the enzyme shows substrate specificity for 3beta-hydroxysteroids. No activity with cholic acid and cholesteryl linoleate Rhodococcus sp. ?
-
?
additional information the enzyme shows substrate specificity for 3beta-hydroxysteroids. No activity with cholic acid and cholesteryl linoleate Rhodococcus sp. PKPD-CL ?
-
?

Subunits

Subunits Comment Organism
? x * 60000, SDS-PAGE Rhodococcus sp.

Synonyms

Synonyms Comment Organism
ChOx
-
Rhodococcus sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
-
Rhodococcus sp.

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
15 55 activity range, profile overview Rhodococcus sp.

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
40 50 purified enzyme, about 80% reduction of activity after 100 days Rhodococcus sp.
60 70 purified enzyme, activity s reduced to 85% and 48% after 10 days and is rapidly reduced during further incubation Rhodococcus sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
-
Rhodococcus sp.

pH Range

pH Minimum pH Maximum Comment Organism
3 13 activity range, profile overview Rhodococcus sp.

pH Stability

pH Stability pH Stability Maximum Comment Organism
4
-
purified enzyme, loss of 80% activity Rhodococcus sp.
6 8.5 purified enzyme, completely stable at Rhodococcus sp.
9.5
-
purified enzyme, loss of 85% activity Rhodococcus sp.

Cofactor

Cofactor Comment Organism Structure
FAD
-
Rhodococcus sp.

pI Value

Organism Comment pI Value Maximum pI Value
Rhodococcus sp. isoelectric focusing
-
9.5

General Information

General Information Comment Organism
additional information a extracellular halo tolerant, detergent and organic solvent stable alkaline cholesterol oxidase. The enzyme is stable in presence of chemical reagents (NaN3, EDTA), detergents (Tween-80, Tween-20, Triton X-100, sodium cholate) and various organic solvents (isopropanol, ethanol, benzene, chloroform, methanol, toluene, ethyl acetate, butanol and dimethylsulfoxide) Rhodococcus sp.