Cloned (Comment) | Organism |
---|---|
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 |
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 | Comment | Organism |
---|---|---|
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] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.12 | - |
cholesterol | pH 7.5, 37°C, full-length isozyme ChoM2 | Mycolicibacterium neoaurum | |
0.14 | - |
cholesterol | pH 7.5, 37°C, truncated isozyme ChoM2 | Mycolicibacterium neoaurum | |
0.17 | - |
cholesterol | pH 7.5, 37°C | Mycolicibacterium neoaurum | |
0.18 | - |
beta-sitosterol | pH 7.5, 37°C, full-length isozyme ChoM2 | Mycolicibacterium neoaurum | |
0.19 | - |
beta-sitosterol | pH 7.5, 37°C, truncated isozyme ChoM2 | Mycolicibacterium neoaurum | |
0.2 | - |
beta-sitosterol | pH 7.5, 37°C | Mycolicibacterium neoaurum | |
0.2 | - |
dehydroandrosterone | pH 7.5, 37°C, full-length isozyme ChoM2 | Mycolicibacterium neoaurum | |
0.23 | - |
diosgenin | pH 7.5, 37°C | Mycolicibacterium neoaurum | |
0.23 | - |
diosgenin | pH 7.5, 37°C, full-length isozyme ChoM2 | Mycolicibacterium neoaurum | |
0.24 | - |
dehydroandrosterone | pH 7.5, 37°C | Mycolicibacterium neoaurum | |
0.25 | - |
dehydroandrosterone | pH 7.5, 37°C, truncated isozyme ChoM2 | Mycolicibacterium neoaurum | |
0.26 | - |
diosgenin | pH 7.5, 37°C, truncated isozyme ChoM2 | Mycolicibacterium neoaurum | |
0.33 | - |
pregnenolone | pH 7.5, 37°C, full-length isozyme ChoM2 | Mycolicibacterium neoaurum | |
0.35 | - |
pregnenolone | pH 7.5, 37°C, truncated isozyme ChoM2 | Mycolicibacterium neoaurum | |
0.38 | - |
pregnenolone | pH 7.5, 37°C | Mycolicibacterium neoaurum |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cell envelope | - |
Mycolicibacterium neoaurum | 30313 | - |
cell envelope | ChoM1 is mainly associated with the cell envelope | Mycolicibacterium neoaurum | 30313 | - |
extracellular | - |
Mycolicibacterium neoaurum | - |
- |
extracellular | ChooM2 is mainly extracellular and soluble | Mycolicibacterium neoaurum | - |
- |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
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 |
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 |
64300 | - |
x * 64300, recombinant His-tagged isozyme ChoM1, SDS-PAGE | Mycolicibacterium neoaurum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
3beta-chlorocholest-5-ene + O2 | Mycolicibacterium neoaurum | low activity | ? + H2O2 | - |
? | |
androst-4-ene-3,17-dione + O2 | Mycolicibacterium neoaurum | - |
? + H2O2 | - |
? | |
beta-sitosterol + O2 | Mycolicibacterium neoaurum | - |
? + H2O2 | - |
? | |
cholest-4-en-3-one + O2 | Mycolicibacterium neoaurum | - |
? + H2O2 | - |
? | |
cholesterol + O2 | Mycolicibacterium neoaurum | - |
cholest-5-en-3-one + H2O2 | - |
? | |
dehydroandrosterone + O2 | Mycolicibacterium neoaurum | - |
? + H2O2 | - |
? | |
diosgenin + O2 | Mycolicibacterium neoaurum | - |
? + H2O2 | - |
? | |
pregnenolone + O2 | Mycolicibacterium neoaurum | - |
? + H2O2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycolicibacterium neoaurum | H9NJ43 | isozyme ChoM1 | - |
Mycolicibacterium neoaurum | H9NJ52 | isozyme ChoM2; several strains | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.1 | - |
purified full-length isozyme ChoM2, substrate pregnenolone, pH 7.5, 37°C | Mycolicibacterium neoaurum |
0.12 | - |
purified truncated isozyme ChoM2, substrate pregnenolone, pH 7.5, 37°C | Mycolicibacterium neoaurum |
0.13 | - |
purified isozyme ChoM1, substrate pregnenolone, pH 7.5, 37°C | Mycolicibacterium neoaurum |
0.16 | - |
purified isozyme ChoM1, substrate dehydroandrosterone, pH 7.5, 37°C | Mycolicibacterium neoaurum |
0.17 | - |
purified truncated isozyme ChoM2, substrate dehydroandrosterone, pH 7.5, 37°C | Mycolicibacterium neoaurum |
0.19 | - |
purified full-length isozyme ChoM2, substrate dehydroandrosterone, pH 7.5, 37°C | Mycolicibacterium neoaurum |
0.19 | - |
purified isozyme ChoM1, substrate diosgenin, pH 7.5, 37°C | Mycolicibacterium neoaurum |
0.21 | - |
purified full-length isozyme ChoM2, substrate diosgenin, pH 7.5, 37°C | Mycolicibacterium neoaurum |
0.22 | - |
purified isozyme ChoM1, substrate beta-sitosterol, pH 7.5, 37°C | Mycolicibacterium neoaurum |
0.23 | - |
purified isozyme ChoM1, substrate cholesterol, pH 7.5, 37°C | Mycolicibacterium neoaurum |
0.25 | - |
purified full-length isozyme ChoM2, substrate beta-sitosterol, pH 7.5, 37°C | Mycolicibacterium neoaurum |
0.26 | - |
purified truncated isozyme ChoM2, substrate diosgenin, pH 7.5, 37°C | Mycolicibacterium neoaurum |
0.28 | - |
purified full-length isozyme ChoM2, substrate cholesterol, pH 7.5, 37°C | Mycolicibacterium neoaurum |
0.33 | - |
purified truncated isozyme ChoM2, substrate beta-sitosterol, pH 7.5, 37°C | Mycolicibacterium neoaurum |
0.42 | - |
purified truncated isozyme ChoM2, substrate cholesterol, pH 7.5, 37°C | Mycolicibacterium neoaurum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3beta-chlorocholest-5-ene + O2 | - |
Mycolicibacterium neoaurum | ? + H2O2 | - |
? | |
3beta-chlorocholest-5-ene + O2 | low activity | Mycolicibacterium neoaurum | ? + H2O2 | - |
? | |
androst-4-ene-3,17-dione + O2 | - |
Mycolicibacterium neoaurum | ? + H2O2 | - |
? | |
beta-sitosterol + O2 | - |
Mycolicibacterium neoaurum | ? + H2O2 | - |
? | |
cholest-4-en-3-one + O2 | - |
Mycolicibacterium neoaurum | ? + H2O2 | - |
? | |
cholesterol + O2 | - |
Mycolicibacterium neoaurum | cholest-5-en-3-one + H2O2 | - |
? | |
dehydroandrosterone + O2 | - |
Mycolicibacterium neoaurum | ? + H2O2 | - |
? | |
diosgenin + O2 | - |
Mycolicibacterium neoaurum | ? + H2O2 | - |
? | |
pregnenolone + O2 | - |
Mycolicibacterium neoaurum | ? + H2O2 | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 57900, recombinant His-tagged isozyme ChoM2, SDS-PAGE x * 55700, recombinant His-tagged isozyme ChoM2 lacking the signal peptide, SDS-PAGE | Mycolicibacterium neoaurum |
? | x * 64300, recombinant His-tagged isozyme ChoM1, SDS-PAGE | Mycolicibacterium neoaurum |
Synonyms | Comment | Organism |
---|---|---|
CHO | - |
Mycolicibacterium neoaurum |
ChoM1 | - |
Mycolicibacterium neoaurum |
ChoM2 | - |
Mycolicibacterium neoaurum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Mycolicibacterium neoaurum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Mycolicibacterium neoaurum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Mycolicibacterium neoaurum |
General Information | Comment | Organism |
---|---|---|
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 |
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 |
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 |
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 |