Recombinant Extracellular Cholesterol Oxidase from Nocardioides simplex

Фокина, В. В.; Karpov, Mikhail V.; Kollerov, Vyacheslav; Bragin, Eugeny Y.; Epiktetov, Dmitry O.; Свиридов, А. В.; Kazantsev, A. V.; Шутов, А. А.; Donova, Marina V.

doi:10.1134/s0006297922090048

Cited by 3 publications

(1 citation statement)

References 33 publications

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“…It was previously reported that the oxidation of sterols can be catalysed by extracellular bacterial oxidases [5,15,16]. The study of the cell-free culture liquid of the M. smegmatis CHE strain did not reveal cholesterol oxidase activity (Figure 4).…”

Section: Oxidation Of Sterols By Mutant Mycolicibacteriamentioning

confidence: 76%

Creation and Characterization of Mycolicibacterium Smegmatis mc²155 with Deletions in Genes Encoding Sterol Oxidation Enzymes

et al. 2023

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The fast-growing saprotrophic strain Mycolicibacterium smegmatis mc2155 is capable of utilizing plant and animal sterols and can be used for creation of genetically engineered strains producing biologically active steroids. Oxidation of the 3β-hydroxyl group and Δ5(6)→Δ4(5) double bond isomerization followed by formation of stenones from sterols are considered as the initial stage of steroid catabolism in some actinobacteria. The study of the mechanism of steroid nucleus 3β-hydroxyl group oxidation is relevant for the creation of a method of the microbiological production of valuable 3β-hydroxy-5-en-steroids. A mutant strain of M. smegmatis with deletions in three genes (MSMEG_1604, MSMEG_5228 and MSMEG_5233) encoding known enzymes exhibiting 3β-hydroxysteroid dehydrogenase activity was constructed by homologous recombination coupled with double selection. The resulting mutant retained macromorphological properties and the ability to convert cholesterol. 3-Keto-4-en-steroids were found among the sterol catabolism intermediates. Experimentally obtained data indicate the presence of a previously undetected intracellular enzyme that performs the function of 3β-hydroxysteroid dehydrogenase/Δ5(6)→Δ4(5) isomerase.

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Section: Oxidation Of Sterols By Mutant Mycolicibacteriamentioning

confidence: 76%

Creation and Characterization of Mycolicibacterium Smegmatis mc²155 with Deletions in Genes Encoding Sterol Oxidation Enzymes

et al. 2023

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Current Trends and Perspectives in Microbial Bioconversions of Steroids

Donova

2023

Methods in Molecular Biology

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Insight into Different Stages of Steroid Degradation in Thermophilic Saccharopolyspora hirsuta VKM Ac-666T Strain

Lobastova

Фокина

Pozdnyakova-Filatova

et al. 2022

IJMS

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Steroids are abundant molecules in nature, and various microorganisms evolved to utilize steroids. Thermophilic actinobacteria play an important role in such processes. However, very few thermophiles have so far been reported capable of degrading or modifying natural sterols. Recently, genes putatively involved in the sterol catabolic pathway have been revealed in the moderately thermophilic actinobacterium Saccharopolyspora hirsuta VKM Ac-666T, but peculiarities of strain activity toward sterols are still poorly understood. S. hirsuta catalyzed cholesterol bioconversion at a rate significantly inferior to that observed for mesophilic actinobacteria (mycobacteria and rhodococci). Several genes related to different stages of steroid catabolism increased their expression in response to cholesterol as was shown by transcriptomic studies and verified by RT–qPCR. Sequential activation of genes related to the initial step of cholesterol side chain oxidation (cyp125) and later steps of steroid core degradation (kstD3, kshA, ipdF, and fadE30) was demonstrated for the first time. The activation correlates with a low cholesterol conversion rate and intermediate accumulation by the strain. The transcriptomic analyses revealed that the genes involved in sterol catabolism are linked functionally, but not transcriptionally. The results contribute to the knowledge on steroid catabolism in thermophilic actinobacteria and could be used at the engineering of microbial catalysts.

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Recombinant Extracellular Cholesterol Oxidase from Nocardioides simplex

Cited by 3 publications

References 33 publications

Creation and Characterization of Mycolicibacterium Smegmatis mc²155 with Deletions in Genes Encoding Sterol Oxidation Enzymes

Creation and Characterization of Mycolicibacterium Smegmatis mc²155 with Deletions in Genes Encoding Sterol Oxidation Enzymes

Current Trends and Perspectives in Microbial Bioconversions of Steroids

Insight into Different Stages of Steroid Degradation in Thermophilic Saccharopolyspora hirsuta VKM Ac-666T Strain

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Recombinant Extracellular Cholesterol Oxidase from Nocardioides simplex

Cited by 3 publications

References 33 publications

Creation and Characterization of Mycolicibacterium Smegmatis mc2155 with Deletions in Genes Encoding Sterol Oxidation Enzymes

Creation and Characterization of Mycolicibacterium Smegmatis mc2155 with Deletions in Genes Encoding Sterol Oxidation Enzymes

Current Trends and Perspectives in Microbial Bioconversions of Steroids

Insight into Different Stages of Steroid Degradation in Thermophilic Saccharopolyspora hirsuta VKM Ac-666T Strain

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Product

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About

Creation and Characterization of Mycolicibacterium Smegmatis mc²155 with Deletions in Genes Encoding Sterol Oxidation Enzymes

Creation and Characterization of Mycolicibacterium Smegmatis mc²155 with Deletions in Genes Encoding Sterol Oxidation Enzymes