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Literature summary extracted from

  • Yao, K.; Wang, F.; Zhang, H.; Wei, D.
    Identification and engineering of cholesterol oxidases involved in the initial step of sterols catabolism in Mycobacterium neoaurum (2013), Metab. Eng., 15, 75-87.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.3.6 gene choM1, DNA and amino acid sequence determination and analysis, genetic organization, expression of His-tagged isozyme ChoM1 in Escherichia coli strain BL21(DE3), subcloning in Escherichia cli strain DH5alpha, overexpression of isozyme ChoM1 in Mycobacterium neoaurum strain NwIB-01MS Mycolicibacterium neoaurum
1.1.3.6 gene choM2, DNA and amino acid sequence determination and analysis, genetic organization, expression of full-length isozyme ChoM2 and of the isozyme without the putative signal peptide sequence in Escherichia coli strain BL21(DE3), subcloning in Escherichia cli strain DH5alpha, overexpression of isozyme ChoM2 in Mycobacterium neoaurum strain NwIB-R10 Mycolicibacterium neoaurum

Protein Variants

EC Number Protein Variants Comment Organism
1.1.3.6 additional information development of a strategy to overcome the rate-limiting step catalzed by the enzyme by augmenting the activity of cholesterol oxidases in Mycobacterium neoaurum strains to enhance their transformation productivity of sterols to valuable steroids, overview Mycolicibacterium neoaurum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.3.6 0.12
-
 cholesterol pH 7.5, 37°C, full-length isozyme ChoM2 Mycolicibacterium neoaurum
1.1.3.6 0.14
-
 cholesterol pH 7.5, 37°C, truncated isozyme ChoM2 Mycolicibacterium neoaurum
1.1.3.6 0.17
-
 cholesterol pH 7.5, 37°C Mycolicibacterium neoaurum
1.1.3.6 0.18
-
 beta-sitosterol pH 7.5, 37°C, full-length isozyme ChoM2 Mycolicibacterium neoaurum
1.1.3.6 0.19
-
 beta-sitosterol pH 7.5, 37°C, truncated isozyme ChoM2 Mycolicibacterium neoaurum
1.1.3.6 0.2
-
 beta-sitosterol pH 7.5, 37°C Mycolicibacterium neoaurum
1.1.3.6 0.2
-
 dehydroandrosterone pH 7.5, 37°C, full-length isozyme ChoM2 Mycolicibacterium neoaurum
1.1.3.6 0.23
-
 diosgenin pH 7.5, 37°C Mycolicibacterium neoaurum
1.1.3.6 0.23
-
 diosgenin pH 7.5, 37°C, full-length isozyme ChoM2 Mycolicibacterium neoaurum
1.1.3.6 0.24
-
 dehydroandrosterone pH 7.5, 37°C Mycolicibacterium neoaurum
1.1.3.6 0.25
-
 dehydroandrosterone pH 7.5, 37°C, truncated isozyme ChoM2 Mycolicibacterium neoaurum
1.1.3.6 0.26
-
 diosgenin pH 7.5, 37°C, truncated isozyme ChoM2 Mycolicibacterium neoaurum
1.1.3.6 0.33
-
 pregnenolone pH 7.5, 37°C, full-length isozyme ChoM2 Mycolicibacterium neoaurum
1.1.3.6 0.35
-
 pregnenolone pH 7.5, 37°C, truncated isozyme ChoM2 Mycolicibacterium neoaurum
1.1.3.6 0.38
-
 pregnenolone pH 7.5, 37°C Mycolicibacterium neoaurum

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.1.3.6 cell envelope
-
 Mycolicibacterium neoaurum 30313
-
1.1.3.6 cell envelope ChoM1 is mainly associated with the cell envelope Mycolicibacterium neoaurum 30313
-
1.1.3.6 extracellular
-
 Mycolicibacterium neoaurum
-
-
1.1.3.6 extracellular ChooM2 is mainly extracellular and soluble Mycolicibacterium neoaurum
-
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.3.6 55700
-
 x * 57900, recombinant His-tagged isozyme ChoM2, SDS-PAGE x * 55700, recombinant His-tagged isozyme ChoM2 lacking the signal peptide, SDS-PAGE Mycolicibacterium neoaurum
1.1.3.6 57900
-
 x * 57900, recombinant His-tagged isozyme ChoM2, SDS-PAGE x * 55700, recombinant His-tagged isozyme ChoM2 lacking the signal peptide, SDS-PAGE Mycolicibacterium neoaurum
1.1.3.6 64300
-
 x * 64300, recombinant His-tagged isozyme ChoM1, SDS-PAGE Mycolicibacterium neoaurum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.3.6 3beta-chlorocholest-5-ene + O2 Mycolicibacterium neoaurum low activity ? + H2O2
-
 ?
1.1.3.6 androst-4-ene-3,17-dione + O2 Mycolicibacterium neoaurum
-
 ? + H2O2
-
 ?
1.1.3.6 beta-sitosterol + O2 Mycolicibacterium neoaurum
-
 ? + H2O2
-
 ?
1.1.3.6 cholest-4-en-3-one + O2 Mycolicibacterium neoaurum
-
 ? + H2O2
-
 ?
1.1.3.6 cholesterol + O2 Mycolicibacterium neoaurum
-
 cholest-5-en-3-one + H2O2
-
 ?
1.1.3.6 dehydroandrosterone + O2 Mycolicibacterium neoaurum
-
 ? + H2O2
-
 ?
1.1.3.6 diosgenin + O2 Mycolicibacterium neoaurum
-
 ? + H2O2
-
 ?
1.1.3.6 pregnenolone + O2 Mycolicibacterium neoaurum
-
 ? + H2O2
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.3.6 Mycolicibacterium neoaurum H9NJ43 isozyme ChoM1
-
1.1.3.6 Mycolicibacterium neoaurum H9NJ52 isozyme ChoM2; several strains
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.3.6 0.1
-
 purified full-length isozyme ChoM2, substrate pregnenolone, pH 7.5, 37°C Mycolicibacterium neoaurum
1.1.3.6 0.12
-
 purified truncated isozyme ChoM2, substrate pregnenolone, pH 7.5, 37°C Mycolicibacterium neoaurum
1.1.3.6 0.13
-
 purified isozyme ChoM1, substrate pregnenolone, pH 7.5, 37°C Mycolicibacterium neoaurum
1.1.3.6 0.16
-
 purified isozyme ChoM1, substrate dehydroandrosterone, pH 7.5, 37°C Mycolicibacterium neoaurum
1.1.3.6 0.17
-
 purified truncated isozyme ChoM2, substrate dehydroandrosterone, pH 7.5, 37°C Mycolicibacterium neoaurum
1.1.3.6 0.19
-
 purified full-length isozyme ChoM2, substrate dehydroandrosterone, pH 7.5, 37°C Mycolicibacterium neoaurum
1.1.3.6 0.19
-
 purified isozyme ChoM1, substrate diosgenin, pH 7.5, 37°C Mycolicibacterium neoaurum
1.1.3.6 0.21
-
 purified full-length isozyme ChoM2, substrate diosgenin, pH 7.5, 37°C Mycolicibacterium neoaurum
1.1.3.6 0.22
-
 purified isozyme ChoM1, substrate beta-sitosterol, pH 7.5, 37°C Mycolicibacterium neoaurum
1.1.3.6 0.23
-
 purified isozyme ChoM1, substrate cholesterol, pH 7.5, 37°C Mycolicibacterium neoaurum
1.1.3.6 0.25
-
 purified full-length isozyme ChoM2, substrate beta-sitosterol, pH 7.5, 37°C Mycolicibacterium neoaurum
1.1.3.6 0.26
-
 purified truncated isozyme ChoM2, substrate diosgenin, pH 7.5, 37°C Mycolicibacterium neoaurum
1.1.3.6 0.28
-
 purified full-length isozyme ChoM2, substrate cholesterol, pH 7.5, 37°C Mycolicibacterium neoaurum
1.1.3.6 0.33
-
 purified truncated isozyme ChoM2, substrate beta-sitosterol, pH 7.5, 37°C Mycolicibacterium neoaurum
1.1.3.6 0.42
-
 purified truncated isozyme ChoM2, substrate cholesterol, pH 7.5, 37°C Mycolicibacterium neoaurum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.3.6 3beta-chlorocholest-5-ene + O2
-
 Mycolicibacterium neoaurum ? + H2O2
-
 ?
1.1.3.6 3beta-chlorocholest-5-ene + O2 low activity Mycolicibacterium neoaurum ? + H2O2
-
 ?
1.1.3.6 androst-4-ene-3,17-dione + O2
-
 Mycolicibacterium neoaurum ? + H2O2
-
 ?
1.1.3.6 beta-sitosterol + O2
-
 Mycolicibacterium neoaurum ? + H2O2
-
 ?
1.1.3.6 cholest-4-en-3-one + O2
-
 Mycolicibacterium neoaurum ? + H2O2
-
 ?
1.1.3.6 cholesterol + O2
-
 Mycolicibacterium neoaurum cholest-5-en-3-one + H2O2
-
 ?
1.1.3.6 dehydroandrosterone + O2
-
 Mycolicibacterium neoaurum ? + H2O2
-
 ?
1.1.3.6 diosgenin + O2
-
 Mycolicibacterium neoaurum ? + H2O2
-
 ?
1.1.3.6 pregnenolone + O2
-
 Mycolicibacterium neoaurum ? + H2O2
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.3.6 ? x * 57900, recombinant His-tagged isozyme ChoM2, SDS-PAGE x * 55700, recombinant His-tagged isozyme ChoM2 lacking the signal peptide, SDS-PAGE Mycolicibacterium neoaurum
1.1.3.6 ? x * 64300, recombinant His-tagged isozyme ChoM1, SDS-PAGE Mycolicibacterium neoaurum

Synonyms

EC Number Synonyms Comment Organism
1.1.3.6 CHO
-
 Mycolicibacterium neoaurum
1.1.3.6 ChoM1
-
 Mycolicibacterium neoaurum
1.1.3.6 ChoM2
-
 Mycolicibacterium neoaurum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.3.6 37
-
 assay at Mycolicibacterium neoaurum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.3.6 7.5
-
 assay at Mycolicibacterium neoaurum

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.3.6 FAD
-
 Mycolicibacterium neoaurum

General Information

EC Number General Information Comment Organism
1.1.3.6 malfunction the deletion mutant of choM1 completely lose the ability to form colonies on the cholesterol agar, the catabolism of sterols is greatly blocked, phenotype, overview Mycolicibacterium neoaurum
1.1.3.6 metabolism cholesterol oxidase ChoM1 is involved in the oxidation of sterols to sterones, a rate-limiting step in the catabolic pathway of sterols in Mycobacterium neoaurum. The bacterium exhibits definite preference for steroidal substrates possessing the 3-keto-4-ene moiety rather than the 3beta-ol-5-ene moiety Mycolicibacterium neoaurum
1.1.3.6 metabolism cholesterol oxidase ChoM2 is involved in the oxidation of sterols to sterones, a rate-limiting step in the catabolic pathway of sterols in Mycobacterium neoaurum Mycolicibacterium neoaurum
1.1.3.6 physiological function the conversion of sterols from 3beta-ol-5-ene moiety to 3-keto-4-ene moiety, i.e. the transformation of sterols to sterones, is a rate-limiting step in the catabolic pathway of sterols in Mycobacterium neoaurum. The conversion of 3beta-ol-5-ene moiety to 3-keto-4-ene moiety of sterols is indispensible for the catabolism of sterols in strain ATCC25795 Mycolicibacterium neoaurum