<scp>FadD</scp>3 is an acyl‐<scp>CoA</scp> synthetase that initiates catabolism of cholesterol rings <scp>C</scp> and <scp>D</scp> in actinobacteria

Casabon, Israël; Crowe, Adam M.; Liu, Jie; Eltis, Lindsay D.

doi:10.1111/mmi.12095

Cited by 76 publications

(136 citation statements)

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“…Strains and plasmids used in this study are described in Table S1 in the supplemental material. RHA1 (33) derivative strains were cultivated as previously described (14,27). Escherichia coli DH5␣ was used for cloning purposes, and E. coli BL21(DE3) (34), Rosetta(DE3)/pLysS, and Rosetta 2(DE3)/pLysS (EMD Millipore) were used for heterologous expression.…”

Section: Methodsmentioning

confidence: 99%

“…3-Oxo-23,24-bisnorchola-1,4-dien-22-oate (1,4-BNC), 3a␣-H-4␣(3=-propanoate)-7a␤-methylhexahydro-1,5-indanedione (HIP), 1␤(2=-propanoate)-3a␣-H-4␣(3Љ(R)-hydroxy-3Љ-propanoate)-7a␤-methylhexahydro-5-indanone (HHIDP), and 1␤(2=-propanoate)-3a␣-H-4␣(3Љ-propanoate)-7a␤-methylhexahydro-5-indanone (HIDP) were obtained as previously described (14,27,36). 3-Hydroxy-9-oxo-9,10-seco-23,24-bisnorchola-1,3,5(10)-trien-22-oate (HSBNC), 3-hydroxy-9-oxo-9,10-seco-23,24-bisnorchola-1,3,5(10),17-tetren-22-oate (⌬ 17 -HSBNC), 3,4-dihydroxy-9-oxo-9,10-seco-23,24-bisnorchola-1,3,5(10)-trien-22-oate (DHSBNC), and 4,5-9,10-diseco-3-hydroxy-5,9-dioxo-23,24-bisnorchola-1(10),2-dien-4,22-dioate (DSHBNC) were obtained as described elsewhere (A. M. Crowe, I. Casabon, and L. D. Eltis, unpublished data).…”

Section: For Expression Of Fadd19mentioning

confidence: 99%

“…Although there is no report about FadD17 or FadD19 of RHA1, their orthologs from M. tuberculosis have been shown to catalyze the CoA thioesterification of long-chain fatty acids (LCFAs) (28), while FadD19 from Rhodococcus rhodochrous DSM 43269 is involved in degradation of C 24 branched-chain sterols (29). Additionally, FadD3 was recently identified as being essential for the catabolism of cholesterol and cholate rings C and D in RHA1 (14), and disruption of orf18, encoding a homolog of FadD3 in C. testosteroni TA441, abrogates degradation of bile acid rings C and D (30). Finally, fadD19 and fadD3 were among those genes predicted by transposon mapping to be essential for growth of M. tuberculosis on cholesterol (31).…”

mentioning

confidence: 99%

“…strain Chol1 (10)(11)(12). Conversely, cholesterol catabolism has been extensively studied in Actinobacteria, especially M. tuberculosis and Rhodococcus species (13)(14)(15)(16)(17). During microbial degradation of steroids, the side chains are degraded via a process similar to the ␤-oxidation of fatty acids.…”

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Actinobacterial Acyl Coenzyme A Synthetases Involved in Steroid Side-Chain Catabolism

et al. 2014

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Section: For Expression Of Fadd19mentioning

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Actinobacterial Acyl Coenzyme A Synthetases Involved in Steroid Side-Chain Catabolism

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“…Both ChsE1/2 and FadE26/27 are part of the KstR regulon (18), which is consistent with their role in side chain degradation. KstR2 regulation of the triplet cluster fadE31-fadE32-fadE33 suggests that the ␣ 2 ␤ 2 ACADs formed from combinations of FadE31/32/33 are likely involved in the degradation of the C/D rings, but this is yet to be confirmed (19). One set of fadE genes (fadE17 and fadE18) is induced by cholesterol but is controlled by Mce3R (20).…”

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Mycobacterium tuberculosis Cholesterol Catabolism Requires a New Class of Acyl Coenzyme A Dehydrogenase

Voskuil

2013

J Bacteriol

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In this month's issue of the Journal of Bacteriology, Wipperman et al. establish a new category of acyl coenzyme A (acyl-CoA) dehydrogenases (ACADs) that form unique ␣ 2 ␤ 2 heterotetrameric complexes required to catalyze unsaturation reactions in ␤-oxidation of steroid-CoA substrates (1). The described ␣ 2 ␤ 2 ACADs allow Mycobacterium tuberculosis, and perhaps other pathogens, to take advantage of ubiquitous cholesterol as a readily available carbon source.The M. tuberculosis genome contains six sets of ACAD genes (fadE genes) that are regulated by cholesterol, with each set of genes found adjacent to each other within the same operon (1). The current study provides compelling evidence that all of these adjacent cholesterol-regulated genes encode members of a new ␣ 2 ␤ 2 heterotetrameric ACAD class. The Sampson laboratory previously demonstrated that two of these genes, chsE1 and chsE2, encode an ␣ 2 ␤ 2 ACAD required for the dehydrogenation of the cholesterol side chain (2). The current study extends the initial characterization of ChsE1/2 to five additional sets of ␣ 2 ␤ 2 ACADs. Of the six ␣ 2 ␤ 2 ACAD sets, five are induced by cholesterol and likely important for cholesterol catabolism, while one set is repressed by cholesterol and is not likely involved in cholesterol catabolism.Acyl-CoA dehydrogenases were first characterized in the 1950s by Beinert and colleagues as enzymes that catalyze the ␣␤ unsaturation of acyl-CoA thioesters in ␤-oxidation (3, 3a). Prior to the recent studies by the Sampson laboratory, ACADs had only been characterized that catalyze the ␣␤ unsaturation of short-, medium-, and long-chain acyl-CoA thioesters, and all were homotetrameric or homodimeric in structure (4). Thus, the recent report on ChsE1/E2 was the first characterization of an ␣ 2 ␤ 2 heterotetrameric ACAD (2). Homotetrameric ACADs have four active sites with four flavin adenine dinucleotide (FAD) cofactors. In five of the six ␣ 2 ␤ 2 ACADs, only one of the adjacent ACAD subunits contains the catalytic residue required for deprotonation of the acyl-CoA substrate. The authors demonstrate that the ␣ 2 ␤ 2 ACADs have one active site and one FAD per heterodimer instead of two. Five of the six genome adjacent fadE pairs formed only heteromeric complexes when expressed in vitro, while the sixth set, FadE17/18, did not form soluble protein. A control pair of fadE genes that are transcribed in the same operon, but are not adjacent pairs, formed typical homomeric complexes only when expressed in vitro and did not have characteristics of the ␣ 2 ␤ 2 ACAD class. Thus, the ␣ 2 ␤ 2 ACADs are formed from two heterodimers in which each specific monomer is required to form a unique active site.The M. tuberculosis genome contains a disproportionally large number of lipid and fatty acid metabolic genes including 35 members of the fadE family (5). This apparent redundancy in the genome has long been a mystery. The study by Wipperman et al. has unraveled a piece of this mystery by establishing a role for 11 fadE genes that encode six highl...

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A Keto Reductase Involved in Steroid Degradation in Mycolicibacterium neoaurum

Jin

Peng

Tian

et al. 2023

Chemistry & Biodiversity

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Phytosterols can be used by microorganisms as carbon and energy sources and completely degraded into CO 2 and H 2 O. The catabolic pathway of phytosterols was well characterized in many microorganisms. Blocking the steroid core ring degradation by deletions of fadE30 and fadD3 genes, two important steroid intermediates, 3aα-H-4α-(3'-Propionic acid)-5α-hydroxy-7aβ-methylhexahydro-1-indanone-δ-lactone (sitolactone, or HIL) and 3aα-H-4α-(3'-propionic acid)-7aβ-methylhexahydro-1,5-indanedione (HIP) can be accumulated. They are currently used to synthesize nor-steroid drugs with an α-methyl group or without the methyl group at the C 10 -position, such as estrone and norethindrone. In this study, a key gene involved in the bioconversion of HIP to HIL was identified in Mycolicibacterium neoaurum. Through heterologous expression, gene hipR was found to be involved in the reduction of the C 5 keto group of HIP to a hydroxy group, leading to spontaneously lactonization into HIL in vitro. Through gene complementation and knockout, HipR functions were verified and two HIP degradation pathways in vivo were elucidated. The finding of this research facilitated the understanding of the metabolic pathway of sterols, and was directly applied to engineering robust production strains by overexpression or knockout of related genes.

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FadD3 is an acyl‐CoA synthetase that initiates catabolism of cholesterol rings C and D in actinobacteria

Cited by 76 publications

References 63 publications

Actinobacterial Acyl Coenzyme A Synthetases Involved in Steroid Side-Chain Catabolism

Actinobacterial Acyl Coenzyme A Synthetases Involved in Steroid Side-Chain Catabolism

Mycobacterium tuberculosis Cholesterol Catabolism Requires a New Class of Acyl Coenzyme A Dehydrogenase

A Keto Reductase Involved in Steroid Degradation in Mycolicibacterium neoaurum

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