Synthesis of 9α-hydroxysteroids by a Rhodococcus sp.

Datcheva, V. K.; Voishvillo, N. E.; Камерницкий, А. В.; Vlahov, R.; Reshetova, I. G.

doi:10.1016/0039-128x(89)90002-0

Cited by 25 publications

(5 citation statements)

References 2 publications

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“…(Gibson et al 1966 ). The enzyme was discovered both in pathogenic and environmental Actinobacteria such as Mycobacterium (Brzostek et al 2005 ; van der Geize et al 2007 ; Wovcha et al 1978 ), Nocardia (Strijewski 1982 ), Rhodococcus (Datcheva et al 1989 ; Geize et al 2002 ; Petrusma et al 2009 ), Arthrobacter (Dutta et al 1992 ), as well as in proteobacteria such as C. testosteroni ( Horinouchi et al 2012 ). Recently, a wide comparative genomic analysis revealed the presence of KSH genes in new genera such as Acinoplanes , Shewanella , and Pseudoalteromonas (Bergstrand et al 2016 ).…”

Section: Non-heme O 2 -Dependent Hydroxylationmentioning

confidence: 99%

Bacterial steroid hydroxylases: enzyme classes, their functions and comparison of their catalytic mechanisms

Szaleniec

Wojtkiewicz

Bernhardt

et al. 2018

Appl Microbiol Biotechnol

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The steroid superfamily includes a wide range of compounds that are essential for living organisms of the animal and plant kingdoms. Structural modifications of steroids highly affect their biological activity. In this review, we focus on hydroxylation of steroids by bacterial hydroxylases, which take part in steroid catabolic pathways and play an important role in steroid degradation. We compare three distinct classes of metalloenzymes responsible for aerobic or anaerobic hydroxylation of steroids, namely: cytochrome P450, Rieske-type monooxygenase 3-ketosteroid 9α-hydroxylase, and molybdenum-containing steroid C25 dehydrogenases. We analyze the available literature data on reactivity, regioselectivity, and potential application of these enzymes in organic synthesis of hydroxysteroids. Moreover, we describe mechanistic hypotheses proposed for all three classes of enzymes along with experimental and theoretical evidences, which have provided grounds for their formulation. In case of the 3-ketosteroid 9α-hydroxylase, such a mechanistic hypothesis is formulated for the first time in the literature based on studies conducted for other Rieske monooxygenases. Finally, we provide comparative analysis of similarities and differences in the reaction mechanisms utilized by bacterial steroid hydroxylases.

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Section: Non-heme O 2 -Dependent Hydroxylationmentioning

confidence: 99%

Bacterial steroid hydroxylases: enzyme classes, their functions and comparison of their catalytic mechanisms

Szaleniec

Wojtkiewicz

Bernhardt

et al. 2018

Appl Microbiol Biotechnol

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“…No accumulation of 9OHAD from AD was observed with either strain SQ1 or strain RG1. In comparison, Datcheva et al [27] reported the isolation of a Rhodococcus sp. converting AD in 9OHAD with a 70% yield at substrate concentrations of 1^3 g l 31 .…”

Section: Bioconversion Of Ad By Cell-free Extracts and Whole Cells Ofmentioning

confidence: 98%

Unmarked gene deletion mutagenesis of kstD, encoding 3-ketosteroid Δ1-dehydrogenase, in Rhodococcus erythropolis SQ1 using sacB as counter-selectable marker

Geize¹

2001

FEMS Microbiology Letters

131

135

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This paper reports the first method for the construction of unmarked gene deletion mutants in the genus Rhodococcus. Unmarked deletion of the kstD gene, encoding 3-ketosteroid Delta1-dehydrogenase (KSTD1) in Rhodococcus erythropolis SQ1, was achieved using the sacB counter-selection system. Conjugative mobilization of the mutagenic plasmid from Escherichia coli S17-1 to R. erythropolis strain SQ1 was used to avoid its random genomic integration. The kstD gene deletion mutant, designated strain RG1, still possessed about 10% of the KSTD enzyme activity of wild-type and was not affected in its ability to grow on the steroid substrates 4-androstene-3,17-dione (AD) and 9alpha-hydroxy-4-androstene-3,17-dione (9OHAD). Biochemical evidence subsequently was obtained for the presence of a second KSTD enzyme (KSTD2) in R. erythropolis SQ1. UV mutants of strain RG1 unable to grow on AD were isolated. One of these mutants, strain RG1-UV29, had lost all KSTD enzyme activity and was also unable to grow on 9OHAD. It stoichiometrically converted AD into 9OHAD in concentrations as high as 20 g x l(-1). The two KSTD enzymes apparently both function in AD and 9OHAD catabolism. These isoenzymes have been inactivated in strain RG1 (KSTD1 negative) and strain RG1-UV29 (KSTD1 and KSTD2 negative), respectively.

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“…KSH activity has been found in many bacterial genera (Martin, 1977; Kieslich, 1985; Mahato and Garai, 1997), e.g. Rhodococcus (Datcheva et al , 1989; van der Geize et al , 2001), Nocardia (Strijewski, 1982), Arthrobacter (Dutta et al , 1992) and Mycobacterium (Wovcha et al , 1978). In previous work, we demonstrated the presence of KSH activity in Rhodococcus erythropolis strain SQ1: inactivation of both kstD and kstD2 , encoding two 3‐ketosteroid Δ1‐dehydrogenase (KSTD) isoenzymes (KSTD1 and KSTD2 respectively) of R. erythropolis SQ1, results in the selective 9α‐hydroxylation of AD, producing >90% 9OHAD (van der Geize et al , 2001).…”

Section: Introductionmentioning

confidence: 99%

Molecular and functional characterization of kshA and kshB, encoding two components of 3‐ketosteroid 9α‐hydroxylase, a class IA monooxygenase, in Rhodococcus erythropolis strain SQ1

Geize

Hessels

Gerwen

et al. 2002

Molecular Microbiology

112

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Summary9a-Hydroxylation of 4-androstene-3,17-dione (AD) and 1,4-androstadiene-3,17-dione (ADD) is catalysed by 3-ketosteroid 9a-hydroxylase (KSH), a key enzyme in microbial steroid catabolism. Very limited knowledge is presently available on the KSH enzyme. Here, we report for the first time the identification and molecular characterization of genes encoding KSH activity. The kshA and kshB genes, encoding KSH in Rhodococcus erythropolis strain SQ1, were cloned by functional complementation of mutant strains blocked in AD(D) 9a-hydroxylation. Analysis of the deduced amino acid sequences of kshA and kshB showed that they contain domains typically conserved in class IA terminal oxygenases and class IA oxygenase reductases respectively. By definition, class IA oxygenases are made up of two components, thus classifying the KSH enzyme system in R. erythropolis strain SQ1 as a two-component class IA monooxygenase composed of KshA and KshB. Unmarked in frame gene deletion mutants of parent strain R. erythropolis SQ1, designated strains RG2 (kshA mutant) and RG4 (kshB mutant), were unable to grow on steroid substrates AD(D), whereas growth on 9a-hydroxy-4-androstene-3,17-dione (9OHAD) was not affected. Incubation of these mutant strains with AD resulted in the accumulation of ADD (30-50% conversion), confirming the involvement of KshA and KshB in AD(D) 9a-hydroxylation. Strain RG4 was also impaired in sterol degradation, suggesting a dual role for KshB in both sterol and steroid degradation.

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Synthesis of 9α-hydroxysteroids by a Rhodococcus sp.

Cited by 25 publications

References 2 publications

Bacterial steroid hydroxylases: enzyme classes, their functions and comparison of their catalytic mechanisms

Bacterial steroid hydroxylases: enzyme classes, their functions and comparison of their catalytic mechanisms

Unmarked gene deletion mutagenesis of kstD, encoding 3-ketosteroid Δ1-dehydrogenase, in Rhodococcus erythropolis SQ1 using sacB as counter-selectable marker

Molecular and functional characterization of kshA and kshB, encoding two components of 3‐ketosteroid 9α‐hydroxylase, a class IA monooxygenase, in Rhodococcus erythropolis strain SQ1

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