Draft Genome Sequence of the Moderately Thermophilic Actinobacterial Steroid-Transforming Saccharopolyspora hirsuta subsp.
            <i>hirsuta</i>
            Strain VKM Ac-666
            <sup>T</sup>

Lobastova, Tatyana G.; Фокина, В. В.; Bragin, Eugeny Y.; Shtratnikova, Victoria; Starodumova, Irina P.; Tarlachkov, Sergey V.; Donova, Marina V.

doi:10.1128/mra.01327-19

Cited by 3 publications

(4 citation statements)

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“…ChOs are most likely involved in 3β-ol-5-ene-moiety modification in S. hirsuta since no candidate genes coding for 3β-HSDs were found in Ac-666 T [25]. Two candidate cho genes, choD F1721_14655 and choE F1721_09795, were revealed in Ac-666 T .…”

Section: Cholesterol Oxidases (Chos)mentioning

confidence: 99%

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Steroid Metabolism in Thermophilic Actinobacterium Saccharopolyspora hirsuta VKM Ac-666T

et al. 2021

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The application of thermophilic microorganisms opens new prospects in steroid biotechnology, but little is known to date on steroid catabolism by thermophilic strains. The thermophilic strain Saccharopolyspora hirsuta VKM Ac-666T has been shown to convert various steroids and to fully degrade cholesterol. Cholest-4-en-3-one, cholesta-1,4-dien-3-one, 26-hydroxycholest-4-en-3-one, 3-oxo-cholest-4-en-26-oic acid, 3-oxo-cholesta-1,4-dien-26-oic acid, 26-hydroxycholesterol, 3β-hydroxy-cholest-5-en-26-oic acid were identified as intermediates in cholesterol oxidation. The structures were confirmed by 1H and 13C-NMR analyses. Aliphatic side chain hydroxylation at C26 and the A-ring modification at C3, which are putatively catalyzed by cytochrome P450 monooxygenase CYP125 and cholesterol oxidase, respectively, occur simultaneously in the strain and are followed by cascade reactions of aliphatic sidechain degradation and steroid core destruction via the known 9(10)-seco-pathway. The genes putatively related to the sterol and bile acid degradation pathways form three major clusters in the S. hirsuta genome. The sets of the genes include the orthologs of those involved in steroid catabolism in Mycobacterium tuberculosis H37Rv and Rhodococcus jostii RHA1 and related actinobacteria. Bioinformatics analysis of 52 publicly available genomes of thermophilic bacteria revealed only seven candidate strains that possess the key genes related to the 9(10)-seco pathway of steroid degradation, thus demonstrating that the ability to degrade steroids is not widespread among thermophilic bacteria.

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Section: Cholesterol Oxidases (Chos)mentioning

confidence: 99%

“…The moderately thermophilic Saccharopolyspora hirsuta VKM Ac-666 T [22] is capable of transforming various steroids, such as lithocholic acid (LCA) [23], dehydroepiandrosterone, androstenedione, and 3β,7(α/β)-dihydroxy-5-ene-D-homo-lactones [24]. Recently, the Ac-666 T genome has been sequenced and preliminary annotated [25].…”

Section: Introductionmentioning

confidence: 99%

Steroid Metabolism in Thermophilic Actinobacterium Saccharopolyspora hirsuta VKM Ac-666T

et al. 2021

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“…Cholesterol oxidases are most likely involved in the 3β-hydroxyl group dehydrogenation and ∆ 5 →∆ 4isomerization (I→II, VII→IV) in S. hirsuta since no candidate genes responsible for the 3β-hydroxysteroid dehydrogenase (3-HSD) were found in the Ac-666 T genome (Lobastova et al 2020). Two candidate cho genes: choD F1721_14655 and choE F1721_09795 encoding the cholesterol oxidases were revealed in the genome and both are situated out of the clusters related to cholesterol catabolism in Ac-666 T .…”

Section: Chos and 3-hsdsmentioning

confidence: 99%

“…hirsuta VKM Ac-666 T originated from the sugarcane bagasse (Lacey and Goodfellow 1975) is able to transform lithocholic acid (Kollerov et al 2013) and steroid compounds (dehydroepiandrosterone, androstenedione, 3β,7(α/β)-dihydroxy-5-ene-Dhomo-lactones) (Lobastova et al 2019). Recently, the Ac-666 T whole genome was sequenced and its preliminary bioinformatics analysis has been performed (Lobastova et al 2020).…”

Section: Read Full License Introductionmentioning

confidence: 99%

Steroid Metabolism In Thermophilic Actinobacteria Saccharopolyspora Hirsuta Subsp. Hirsuta VKM Ac-666T

Lobastova

Фокина

Tarlachkov

et al. 2021

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Application of thermophile microorganisms opens new prospects in steroid biotechnology, however little is known on steroid catabolism by the thermophile strains.The thermophilic Saccharopolyspora hirsuta subsp. hirsuta strain VKM Ac-666T is capable of structural modification of different steroids, and fully degrades cholesterol. The intermediates of the cholesterol degradation pathway were identified as cholest-4-en-3-one, cholesta-1,4-dien-3-one, 26-hydroxycholest-4-en-3-one, 3-oxo-cholest-4-en-26-oic acid, 3-oxo-cholesta-1,4-dien-26-oic acid, 26-hydroxycholesterol, 3β-hydroxy-cholest-5-en-26-oic acid by MS, and H1- and C13-NMR analyses. The data evidence sterol degradation by the strain occurs simultaneously through the aliphatic side chain hydroxylation at C26 and the A-ring modification that are putatively catalyzed by cytochrome P450 monooxygenase CYP125 and cholesterol oxidase, respectively.The genes orthologous to those related to the sterol side chain degradation, steroid core rings A/B and C/D disruption and the steroid uptake were revealed. Most of the genes related to steroid degradation are grouped in three clusters. The sets of the genes putatively involved in steroid catabolism and peculiarities of their organization in S. hirsuta are discussed.Despite steroids abundancy in the environments, the ability to degrade them is not widespread among thermophilic bacteria as follows from the bioinformatic analysis of 52 publicly available genomes. Only seven candidate strains were revealed to possess the key genes related to the only known 9(10)-seco pathway of steroid degradation.The results contribute to the knowledge on diversity of microbial steroid degraders, the features of sterol catabolism by thermophilic actinobacteria and could be useful for application in the pharmaceutical and environmental biotechnology.

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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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Draft Genome Sequence of the Moderately Thermophilic Actinobacterial Steroid-Transforming Saccharopolyspora hirsuta subsp. hirsuta Strain VKM Ac-666 ^T

Abstract: The draft genome sequence of the type strain Saccharopolyspora hirsuta subsp. hirsuta VKM Ac-666 was sequenced. This moderately thermophilic actinobacterial strain of sugarcane bagasse origin is able to transform different steroid substrates.

Cited by 3 publications

References 11 publications

Steroid Metabolism in Thermophilic Actinobacterium Saccharopolyspora hirsuta VKM Ac-666T

Steroid Metabolism in Thermophilic Actinobacterium Saccharopolyspora hirsuta VKM Ac-666T

Steroid Metabolism In Thermophilic Actinobacteria Saccharopolyspora Hirsuta Subsp. Hirsuta VKM Ac-666T

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

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Draft Genome Sequence of the Moderately Thermophilic Actinobacterial Steroid-Transforming Saccharopolyspora hirsuta subsp. hirsuta Strain VKM Ac-666 T

Abstract: The draft genome sequence of the type strain Saccharopolyspora hirsuta subsp. hirsuta VKM Ac-666 was sequenced. This moderately thermophilic actinobacterial strain of sugarcane bagasse origin is able to transform different steroid substrates.

Cited by 3 publications

References 11 publications

Steroid Metabolism in Thermophilic Actinobacterium Saccharopolyspora hirsuta VKM Ac-666T

Steroid Metabolism in Thermophilic Actinobacterium Saccharopolyspora hirsuta VKM Ac-666T

Steroid Metabolism In Thermophilic Actinobacteria Saccharopolyspora Hirsuta Subsp. Hirsuta VKM Ac-666T

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

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Draft Genome Sequence of the Moderately Thermophilic Actinobacterial Steroid-Transforming Saccharopolyspora hirsuta subsp. hirsuta Strain VKM Ac-666 ^T