Microbiological synthesis of stereoisomeric 7(α/β)-hydroxytestololactones and 7(α/β)-hydroxytestolactones

Tarlachkov

et al. 2021

Preprint

Self Cite

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.

Section: Steroid Nucleus Degradationmentioning

confidence: 67%

Section: Read Full License Introductionmentioning

confidence: 99%

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

Lobastova

Tarlachkov

et al. 2021

Preprint

Self Cite

“…Steroids with 3-oxo-4-ene moiety were visualized under UV light (254 nm) using hemiscope CN-15MC UV Darkroom (Vilber Lourmat, Collégien, France). LCA metabolites were assayed as described earlier [ 22 , 26 ].…”

Section: Methodsmentioning

confidence: 99%

Genome-Wide Transcriptome Profiling Provides Insight on Cholesterol and Lithocholate Degradation Mechanisms in Nocardioides simplex VKM Ac-2033D

Shtratnikova

Schelkunov

et al. 2020

Genes

Steroid microbial degradation plays a significant ecological role for biomass decomposition and removal/detoxification of steroid pollutants. In this study, the initial steps of cholesterol degradation and lithocholate bioconversion by a strain with enhanced 3-ketosteroid dehydrogenase (3-KSD) activity, Nocardioides simplex VKM Ac-2033D, were studied. Biochemical, transcriptomic, and bioinformatic approaches were used. Among the intermediates of sterol sidechain oxidation cholest-5-en-26-oic acid and 3-oxo-cholesta-1,4-dien-26-oic acid were identified as those that have not been earlier reported for N. simplex and related species. The transcriptomic approach revealed candidate genes of cholesterol and lithocholic acid (LCA) catabolism by the strain. A separate set of genes combined in cluster and additional 3-ketosteroid Δ1-dehydrogenase and 3-ketosteroid 9α-hydroxylases that might be involved in LCA catabolism were predicted. Bioinformatic calculations based on transcriptomic data showed the existence of a previously unknown transcription factor, which regulates cholate catabolism gene orthologs. The results contribute to the knowledge on diversity of steroid catabolism regulation in actinobacteria and might be used at the engineering of microbial catalysts for ecological and industrial biotechnology.

“…hirsuta were originally isolated from spontaneously heated sugarcane bagasse (5). The strain utilized lithocholic acid (6), transformed 3␤-ol-5-ene steroids to their corresponding 3-keto-4ene derivatives, and catalyzed 1(2)-dehydrogenation as it was demonstrated for dehydroepiandrosterone and 7-hydroxylated steroid D-lactones (7); however, the molecular mechanisms of its steroid-transforming activities have not been studied yet. Here, we report a whole-genome sequence of the strain.…”

mentioning

confidence: 99%

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

Lobastova

Microbiol Resour Announc

Bragin

et al. 2020