Dataset Information

Complete Genome Sequence of Mycobacterium sp. Strain VKM Ac-1817D, Capable of Producing 9?-Hydroxy-androst-4-ene-3,17-dione from Phytosterol.

ABSTRACT: Mycobacterium sp. strain VKM Ac-1817D is capable of converting phytosterol into 9?-hydroxy androst-4-ene-3,17-dione (9-OH-AD), which is a valuable intermediate for the steroid pharmaceutical industry. Here, a complete genome sequence of the strain is reported. The genome consists of a single circular 6,324,222-bp chromosome with a G+C content of 66.2% and encodes approximately 6,000 CDSs, 54 tRNAs, and 6 rRNAs.

SUBMITTER: Shtratnikova VY

PROVIDER: S-EPMC4319502 | biostudies-literature | 2015 Jan

REPOSITORIES: biostudies-literature

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Complete Genome Sequence of Mycobacterium sp. Strain VKM Ac-1817D, Capable of Producing 9α-Hydroxy-androst-4-ene-3,17-dione from Phytosterol.

Shtratnikova Victoriya Y VY Schelkunov Mikhail I MI Dovbnya Dmitry V DV Pekov Yury A YA Bragin Eugeny Y EY Ashapkin Vasily V VV Donova Marina V MV

Genome announcements 20150129 1

Mycobacterium sp. strain VKM Ac-1817D is capable of converting phytosterol into 9α-hydroxy androst-4-ene-3,17-dione (9-OH-AD), which is a valuable intermediate for the steroid pharmaceutical industry. Here, a complete genome sequence of the strain is reported. The genome consists of a single circular 6,324,222-bp chromosome with a G+C content of 66.2% and encodes approximately 6,000 CDSs, 54 tRNAs, and 6 rRNAs. ...[more]

PMID: 25635031

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Project description:BackgroundAerobic side chain degradation of phytosterols by actinobacteria is the basis for the industrial production of androstane steroids which are the starting materials for the synthesis of steroid hormones. A native strain of Mycobacterium sp. VKM Ac-1817D effectively produces 9α-hydroxyandrost-4-ene-3,17-dione (9-OH-AD) from phytosterol, but also is capable of slow steroid core degradation. However, the set of the genes with products that are involved in phytosterol oxidation, their organisation and regulation remain poorly understood.ResultsHigh-throughput sequencing of the global transcriptomes of the Mycobacterium sp. VKM Ac-1817D cultures grown with or without phytosterol was carried out. In the presence of phytosterol, the expression of 260 genes including those related to steroid catabolism pathways significantly increased. Two of the five genes encoding the oxygenase unit of 3-ketosteroid-9α-hydroxylase (kshA) were highly up-regulated in response to phytosterol (55- and 25-fold, respectively) as well as one of the two genes encoding its reductase subunit (kshB) (40-fold). Only one of the five putative genes encoding 3-ketosteroid-∆1-dehydrogenase (KstD_1) was up-regulated in the presence of phytosterol (61-fold), but several substitutions in the conservative positions of its product were revealed. Among the genes over-expressed in the presence of phytosterol, several dozen genes did not possess binding sites for the known regulatory factors of steroid catabolism. In the promoter regions of these genes, a regularly occurring palindromic motif was revealed. The orthologue of TetR-family transcription regulator gene Rv0767c of M. tuberculosis was identified in Mycobacterium sp. VKM Ac-1817D as G155_05115.ConclusionsHigh expression levels of the genes related to the sterol side chain degradation and steroid 9α-hydroxylation in combination with possible defects in KstD_1 may contribute to effective 9α-hydroxyandrost-4-ene-3,17-dione accumulation from phytosterol provided by this biotechnologically relevant strain. The TetR-family transcription regulator gene G155_05115 presumably associated with the regulation of steroid catabolism. The results are of significance for the improvement of biocatalytic features of the microbial strains for the steroid industry.

Project description:BackgroundBacterial degradation/transformation of steroids is widely investigated to create biotechnologically relevant strains for industrial application. The strain of Nocardioides simplex VKM Ac-2033D is well known mainly for its superior 3-ketosteroid ?1-dehydrogenase activity towards various 3-oxosteroids and other important reactions of sterol degradation. However, its biocatalytic capacities and the molecular fundamentals of its activity towards natural sterols and synthetic steroids were not fully understood. In this study, a comparative investigation of the genome-wide transcriptome profiling of the N. simplex VKM Ac-2033D grown on phytosterol, or in the presence of cortisone 21-acetate was performed with RNA-seq.ResultsAlthough the gene patterns induced by phytosterol generally resemble the gene sets involved in phytosterol degradation pathways in mycolic acid rich actinobacteria such as Mycolicibacterium, Mycobacterium and Rhodococcus species, the differences in gene organization and previously unreported genes with high expression level were revealed. Transcription of the genes related to KstR- and KstR2-regulons was mainly enhanced in response to phytosterol, and the role in steroid catabolism is predicted for some dozens of the genes in N. simplex. New transcription factors binding motifs and new candidate transcription regulators of steroid catabolism were predicted in N. simplex. Unlike phytosterol, cortisone 21-acetate does not provide induction of the genes with predicted KstR and KstR2 sites. Superior 3-ketosteroid-?1-dehydrogenase activity of N. simplex VKM Ac-2033D is due to the kstDs redundancy in the genome, with the highest expression level of the gene KR76_27125 orthologous to kstD2, in response to cortisone 21-acetate. The substrate spectrum of N. simplex 3-ketosteroid-?1-dehydrogenase was expanded in this study with progesterone and its 17?-hydroxylated and 11?,17?-dihydroxylated derivatives, that effectively were 1(2)-dehydrogenated in vivo by the whole cells of the N. simplex VKM Ac-2033D.ConclusionThe results contribute to the knowledge of biocatalytic features and diversity of steroid modification capabilities of actinobacteria, defining targets for further bioengineering manipulations with the purpose of expansion of their biotechnological applications.

Dataset Information

Complete Genome Sequence of Mycobacterium sp. Strain VKM Ac-1817D, Capable of Producing 9?-Hydroxy-androst-4-ene-3,17-dione from Phytosterol.

Publications

Complete Genome Sequence of Mycobacterium sp. Strain VKM Ac-1817D, Capable of Producing 9α-Hydroxy-androst-4-ene-3,17-dione from Phytosterol.

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