Two moles of O2 consumption and one mole of H2O2 formation during cholesterol peroxidation with cholesterol oxidase from <i>Pseudomonas</i> sp. strain ST-200

DOUKYU, Noriyuki; Aono, Rikizo

doi:10.1042/bj3410621

Cited by 13 publications

(5 citation statements)

References 10 publications

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“…The R f (factor rate) value and retention time of product 1 matched exactly with those of cholestenone. However, product 2 presented an R f that could correspond to 6β‐hydroperoxy‐cholest‐4‐en‐3‐one, a secondary product resulting from an alternative activity of some ChOxs (Doukyu and Aono, 1999; Doukyu et al ., 2008). In order to confirm the structure of product 2, the sample was extracted and analysed by liquid chromatography/mass spectrometry (LC/MS) as described in Experimental procedures .…”

Section: Resultsmentioning

confidence: 99%

Initial step in the catabolism of cholesterol by Mycobacterium smegmatis mc²155

Uhía

Galán²,

Morales

et al. 2011

Environmental Microbiology

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The first step in the catabolism of cholesterol, i.e. the transformation of cholesterol into cholestenone, has been investigated in Mycobacterium smegmatis. In silico analysis identified the MSMEG_1604 gene encoding a putative protein similar to the ChoD cholesterol oxidase of M. tuberculosis H37Rv (Rv3409c) and the MSMEG_5228 gene coding for a protein similar to the NAD(P)-dependent cholesterol dehydrogenase/isomerase of Nocardia sp. The expression of the MSMEG_5228 gene was inducible by cholesterol whereas the expression of MSMEG_1604 gene was constitutive. When both genes were expressed in Escherichia coli only the MSMEG_5228 protein was active on cholesterol. The function of ChoD-like MSMEG_1604 protein remains to be elucidated, but it does not appear to play a critical role in the mineralization of cholesterol as a MSMEG_1604(-) mutant was not affected in the production of cholestenone. However, a MSMEG_5228(-) mutant showed a drastic reduction in the synthesis of cholestenone. The finding that this mutant was still able to grow in cholesterol, allowed us to demonstrate that the cholesterol-inducible MSMEG_5233 gene encodes an additional cholesterol dehydrogenase/isomerase similar to the AcmA dehydrogenase of Sterolibacterium denitrificans. The observation that the double MSMEG_5228-5233(-) mutant was able to grow in cholesterol suggests that in addition to these enzymes other dehydrogenase/isomerases can also catalyse the first reaction of the cholesterol degradation pathway in M. smegmatis, which is not the limiting step of the process.

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Section: Resultsmentioning

confidence: 99%

Initial step in the catabolism of cholesterol by Mycobacterium smegmatis mc²155

Uhía

Galán²,

Morales

et al. 2011

Environmental Microbiology

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“…Unlike most ChoXs, the VAO/PCMH-type enzymes from Gram-negative bacteria do not catalyze the isomerization of cholest-5-en-3-one to cholest-4-en-3-one and the main product of the reaction is 6hydroperoxycholest-4-en-3-one (Fig. 13) [140]. The exact mechanism by which 6-hydroperoxycholest-4-en-3-one is formed is unclear.…”

Section: Subgroup 11: Cholesterol Oxidasesmentioning

confidence: 99%

“…The exact mechanism by which 6-hydroperoxycholest-4-en-3-one is formed is unclear. It has been proposed that it may be the result of dioxygenase activity of the enzyme, but a mechanism where the oxidation proceeds through enzyme-initiated radical reactions is also a possibility [140,141]. The molecular determinants of which product is formed are unclear and would be an interesting topic for future studies, particularly considering that a crystal structure is available of both the cholest-4-en-3-one-forming VAO/PCMH-type ChoX from B. sterolicum and the 6-hydroperoxycholest-4-en-3-oneforming VAO/PCMH-type ChoX from Chromobacterium sp.…”

Section: Subgroup 11: Cholesterol Oxidasesmentioning

confidence: 99%

The VAO/PCMH flavoprotein family

Ewing

Fraaije

Mattevi

et al. 2017

Archives of Biochemistry and Biophysics

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The VAO/PCMH flavoprotein family consists of structurally homologous flavin-dependent enzymes that catalyze a wide range of chemical reactions. Family members share an architecture consisting of a conserved FAD-binding domain and a more variable substrate-binding domain, which enables varying interactions with a range of substrates while maintaining the same cofactor-binding fold. Here, we provide an overview of the current state of our knowledge on the members of the VAO/PCMH family. Based on a phylogenetic analysis, we divide the family into 11 subgroups. We discuss the properties of these subgroups, focusing on recent developments in terms of the discovery of new family members and mechanistic advances. We also highlight open questions that will provide challenges for future research.

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“…and Chromobacterium sp. DS‐1, oxidize cholesterol mainly to 6β‐hydroperoxy‐cholest‐4‐en‐3‐one and hydrogen peroxide instead of cholestenone (Doukyu and Aono, 1999; Doukyu et al ., 2008; Doukyu, 2009) but this intermediate has not been associated so far to any cholesterol degradation pathway. Recently, a cholesterol inducible ChoX encoding gene ( choG ) has been identified in Rhodococcus sp.…”

Section: Catabolism Of Cholesterolmentioning

confidence: 99%

Catabolism and biotechnological applications of cholesterol degrading bacteria

Garcı́a

Uhía

Galán

2012

Microbial Biotechnology

146

111

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SummaryCholesterol is a steroid commonly found in nature with a great relevance in biology, medicine and chemistry, playing an essential role as a structural component of animal cell membranes. The ubiquity of cholesterol in the environment has made it a reference biomarker for environmental pollution analysis and a common carbon source for different microorganisms, some of them being important pathogens such as Mycobacterium tuberculosis. This work revises the accumulated biochemical and genetic knowledge on the bacterial pathways that degrade or transform this molecule, given that the characterization of cholesterol metabolism would contribute not only to understand its role in tuberculosis but also to develop new biotechnological processes that use this and other related molecules as starting or target materials.

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Two moles of O2 consumption and one mole of H2O2 formation during cholesterol peroxidation with cholesterol oxidase from Pseudomonas sp. strain ST-200

Cited by 13 publications

References 10 publications

Initial step in the catabolism of cholesterol by Mycobacterium smegmatis mc²155

Initial step in the catabolism of cholesterol by Mycobacterium smegmatis mc²155

The VAO/PCMH flavoprotein family

Catabolism and biotechnological applications of cholesterol degrading bacteria

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Two moles of O2 consumption and one mole of H2O2 formation during cholesterol peroxidation with cholesterol oxidase from Pseudomonas sp. strain ST-200

Cited by 13 publications

References 10 publications

Initial step in the catabolism of cholesterol by Mycobacterium smegmatis mc2155

Initial step in the catabolism of cholesterol by Mycobacterium smegmatis mc2155

The VAO/PCMH flavoprotein family

Catabolism and biotechnological applications of cholesterol degrading bacteria

Contact Info

Product

Resources

About

Initial step in the catabolism of cholesterol by Mycobacterium smegmatis mc²155

Initial step in the catabolism of cholesterol by Mycobacterium smegmatis mc²155