Literature DB >> 23474435

Comparative analysis of genes encoding key steroid core oxidation enzymes in fast-growing Mycobacterium spp. strains.

E Yu Bragin1, V Yu Shtratnikova, D V Dovbnya, M I Schelkunov, Yu A Pekov, S G Malakho, O V Egorova, T V Ivashina, S L Sokolov, V V Ashapkin, M V Donova.   

Abstract

A comparative genome analysis of Mycobacterium spp. VKM Ac-1815D, 1816D and 1817D strains used for efficient production of key steroid intermediates (androst-4-ene-3,17-dione, AD, androsta-1,4-diene-3,17-dione, ADD, 9α-hydroxy androst-4-ene-3,17-dione, 9-OH-AD) from phytosterol has been carried out by deep sequencing. The assembled contig sequences were analyzed for the presence putative genes of steroid catabolism pathways. Since 3-ketosteroid-9α-hydroxylases (KSH) and 3-ketosteroid-Δ(1)-dehydrogenase (Δ(1) KSTD) play key role in steroid core oxidation, special attention was paid to the genes encoding these enzymes. At least three genes of Δ(1) KSTD (kstD), five genes of KSH subunit A (kshA), and one gene of KSH subunit B of 3-ketosteroid-9α-hydroxylases (kshB) have been found in Mycobacterium sp. VKM Ac-1817D. Strains of Mycobacterium spp. VKM Ac-1815D and 1816D were found to possess at least one kstD, one kshB and two kshA genes. The assembled genome sequence of Mycobacterium sp. VKM Ac-1817D differs from those of 1815D and 1816D strains, whereas these last two are nearly identical, differing by 13 single nucleotide substitutions (SNPs). One of these SNPs is located in the coding region of a kstD gene and corresponds to an amino acid substitution Lys (135) in 1816D for Ser (135) in 1815D. The findings may be useful for targeted genetic engineering of the biocatalysts for biotechnological application.
Copyright © 2013. Published by Elsevier Ltd.

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Keywords:  2,3-dehydroxyphenyl dioxygenase; 2-enoyl acyl-CoA hydratase; 2-hydroxypenta-2,4-dienoate hydratase; 3,4-dihydroxy-9,10-secoandrosta-1,3,5(10)-triene-9,17-dione 4,5-dioxygenase; 3-hydroxy-9,10-secoandrosta-1,3,5(10)-triene-9,17-dione monooxygenase; 3-hydroxy-9,10-secoandrosta-1,3,5(10)-triene-9,17-dione monooxygenase subunit; 3-ketosteroid-9α-hydroxylase; 3-ketosteroid-Δ(1)-dehydrogenase; 3β-hydroxysteroid-dehydrogenase; 4,5:9,10-diseco-3-hydroxy-5,9,17-trioxoandrosta-1(10),2-diene-4-oate hydrolase; 4-hydroxy-2-oxovalerate aldolase; 9-OH-AD; 9α-hydroxy androst-4-ene-3,17-dione; AD; ADD; Androst-1,4-diene-3,17-dione; Androst-4-ene-3,17-dione; BWA; Broadband-Wheeler Aligner; CTAB; ChoX; ChoX(D,E); EchA19; FAD; FadA5; FadD17; FadD19; FadE26; FadE27; FadE28; Genome sequencing; HSD; HTH-type transcriptional repressor; HsaA; HsaAB; HsaB; HsaC; HsaD; HsaE; HsaF; HsaG; Hsd4A; Hsd4B; KSH; KshA; KshB; KstR; KstR2; Ltp2; Ltp3; Ltp4; Mycobacterium; ORFs; PWM; Phytosterol; SNP; Steroid bioconversion; TesB; YrbE4A; YrbE4B; acetaldehyde dehydrogenase; acetyl-CoA acetyltransferase; acyl-CoA dehydrogenase; acyl-CoA synthetase; acyl-CoA thioesterase II; androst-4-ene-3,17-dione; androsta-1,4-diene-3,17-dione; base pair; bp; cetyl trimethyl ammonium bromide; cholesterol oxidase; enoyl-CoA hydratase; flavin adenine dinucleotide; hydroxysteroid dehydrogenase; integral membrane protein; lipid transfer protein 4 (keto acyl-CoA thiolase); lipid-transfer protein 2; lipid-transfer protein 3 (acetyl-CoA acetyltransferase); open reading frames; position weight matrix; single nucleotide substitution; subunit A of 3-ketosteroid-9α-hydroxylase; subunit B of 3-ketosteroid-9α-hydroxylases; Δ(1) KSTD

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Year:  2013        PMID: 23474435     DOI: 10.1016/j.jsbmb.2013.02.016

Source DB:  PubMed          Journal:  J Steroid Biochem Mol Biol        ISSN: 0960-0760            Impact factor:   4.292


  20 in total

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6.  Engineered 3-Ketosteroid 9α-Hydroxylases in Mycobacterium neoaurum: an Efficient Platform for Production of Steroid Drugs.

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7.  A mutant form of 3-ketosteroid-Δ(1)-dehydrogenase gives altered androst-1,4-diene-3, 17-dione/androst-4-ene-3,17-dione molar ratios in steroid biotransformations by Mycobacterium neoaurum ST-095.

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8.  Genome-wide bioinformatics analysis of steroid metabolism-associated genes in Nocardioides simplex VKM Ac-2033D.

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

Authors:  Victoriya Y Shtratnikova; Mikhail I Schelkunov; Dmitry V Dovbnya; Yury A Pekov; Eugeny Y Bragin; Vasily V Ashapkin; Marina V Donova
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10.  Complete Genome Sequence of Sterol-Transforming Mycobacterium neoaurum Strain VKM Ac-1815D.

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