Steroids, the steroid community, and Upjohn in perspective: a profile of innovation

Hogg, John A.

doi:10.1016/0039-128x(92)90013-y

Cited by 129 publications

(67 citation statements)

References 52 publications

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“…For more than 60 years, bacterial transformation of natural steroid molecules has played an essential role in the biotechnological production of steroid-based pharmaceuticals, such as cortisol derivatives or sex hormones (1)(2)(3). Bacterial steroid metabolism is also very important for the degradation of synthetic steroid pharmaceuticals, which are thought to influence the fertility of animals and humans (4).…”

mentioning

confidence: 99%

The Essential Function of Genes for a Hydratase and an Aldehyde Dehydrogenase for Growth of Pseudomonas sp. Strain Chol1 with the Steroid Compound Cholate Indicates an Aldolytic Reaction Step for Deacetylation of the Side Chain

2013

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In the bacterial degradation of steroid compounds, the enzymes initiating the breakdown of the steroid rings are well known, while the reactions for degrading steroid side chains attached to C-17 are largely unknown. A recent in vitro analysis with Pseudomonas sp. strain Chol1 has shown that the degradation of the C 5 acyl side chain of the C 24 steroid compound cholate involves the C 22 intermediate 7␣,12␣-dihydroxy-3-oxopregna-1,4-diene-20S-carbaldehyde (DHOPDCA) with a terminal aldehyde group. In the present study, candidate genes with plausible functions in the formation and degradation of this aldehyde were identified. All deletion mutants were defective in growth with cholate but could transform it into dead-end metabolites. A mutant with a deletion of the shy gene, encoding a putative enoyl coenzyme A (CoA) hydratase, accumulated the C 24 steroid (22E)-7␣,12␣-dihydroxy-3-oxochola-1,4,22-triene-24-oate (DHOCTO). Deletion of the sal gene, formerly annotated as the steroid ketothiolase gene skt, resulted in the accumulation of 7␣,12␣,22-trihydroxy-3-oxochola-1,4-diene-24-oate (THOCDO). In cell extracts of strain Chol1, THOCDO was converted into DHOPDCA in a coenzyme A-and ATP-dependent reaction. A sad deletion mutant accumulated DHOPDCA, and expression in Escherichia coli revealed that sad encodes an aldehyde dehydrogenase for oxidizing DHOPDCA to the corresponding acid 7␣,12␣-dihydroxy-3-oxopregna-1,4-diene-20-carboxylate (DHOPDC) with NAD ؉ as the electron acceptor. These results clearly show that the degradation of the acyl side chain of cholate proceeds via an aldolytic cleavage of an acetyl residue; they exclude a thiolytic cleavage for this reaction step. Based on these results and on sequence alignments with predicted aldolases from other bacteria, we conclude that the enzyme encoded by sal catalyzes this aldolytic cleavage.

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confidence: 99%

The Essential Function of Genes for a Hydratase and an Aldehyde Dehydrogenase for Growth of Pseudomonas sp. Strain Chol1 with the Steroid Compound Cholate Indicates an Aldolytic Reaction Step for Deacetylation of the Side Chain

2013

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“…For example, hormone steroids, found mainly in livestock runoff and sewage effluent, are problematic due to endocrine disruption in humans and in fish and other wildlife (2,18,33). Alternatively, certain steroids have therapeutic properties and are routinely prescribed to treat a wide variety of diseases and disorders (16,19). Ongoing efforts are aimed at discovering new pharmaceutically useful steroids.…”

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confidence: 99%

Two Transporters Essential for Reassimilation of Novel Cholate Metabolites by Rhodococcus jostii RHA1

et al. 2012

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The bacterial uptake of steroids and their metabolites remains poorly understood. We investigated two transporters associated with cholate catabolism in Rhodococcus jostii RHA1. Reverse transcriptase quantitative-PCR indicated that an ATP-binding cassette (ABC) transporter and a major facilitator superfamily (MFS) transporter were upregulated 16.7-and 174-fold, respectively, during the exponential phase of growth on cholate compared to growth on pyruvate. Gene knockout analysis established that these transporters are required for the reassimilation of distinct metabolites that accumulate during growth on cholate. The ABC transporter, encoded by camABCD, was essential for uptake of 1␤(2=-propanoate)-3a␣-H-4␣(3؆(R)-hydroxy-3؆-propanoate)-7a␤-methylhexahydro-5-indanone and a desaturated analog. The MFS transporter, encoded by camM, was essential for uptake of 3,7(R),12(S)-trihydroxy-9-oxo-9,10-seco-23,24-bisnorchola-1,3,5(10)-trien-22-oate. These metabolites differ from cholate metabolites reported to be excreted by proteobacteria in that they retain an isopropanoyl side chain at C-17. The uptake of these metabolites was necessary for maximal growth on cholate: a ⌬camB mutant lacking the permease component of the ABC transporter and a ⌬camM mutant lacking the MFS transporter grew to 74% and 77%, respectively, of the yield of the wild type. This study demonstrates for the first time the requirement for specific transporters for uptake of cholate metabolites and highlights the importance and complexity of transport processes associated with bacterial steroid catabolism.

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“…In the Upjohn industrial process, an Aspergillus niger strain was used for oxidatively hydroxylating the desired C11-position regioselectively with formation of the C11-a-alcohol 4. 9 This was the wrong diastereomer in an otherwise impressive site-selective CH-activating process. At the time the nature of the actual enzyme was not known, but it was most likely a cytochrome P450 monooxygenase (CYP), enzymes that have since been used in many selective oxidation reactions.…”

Section: Guangyue LImentioning

confidence: 99%

“…9,10 CYPs have large binding pockets, which leads to two basic problems: Small molecules such as C3-C5 alkanes are generally not accepted, while larger ones are often randomly attacked at different positions with formation of undesired mixtures of regio-and stereoisomers. 10 Rational design utilizing site-specific mutagenesis continues to provide progress in this challenging research area, but ongoing studies relying on directed evolution indicate that this approach constitutes the more general and reliable strategy.…”

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confidence: 99%

Enzymatic site-selectivity enabled by structure-guided directed evolution

Wang

Reetz

2017

Chem. Commun.

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Biocatalytic site-selective (regioselective) organic transformations have been practiced for decades, but the traditional limitations of enzymes regarding narrow substrate acceptance and the often observed insufficient degree of selectivity have persisted until recently. With the advent of directed evolution, it is possible to engineer site-selectivity to suit the needs of organic chemists. This review features recent progress in this exciting research area, selected examples involving P450 monooxygenases, halogenases and Baeyer-Villiger monooxygenases being featured for illustrative purposes. The complementary nature of enzymes and man-made catalysts is emphasized.

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Steroids, the steroid community, and Upjohn in perspective: a profile of innovation

Cited by 129 publications

References 52 publications

The Essential Function of Genes for a Hydratase and an Aldehyde Dehydrogenase for Growth of Pseudomonas sp. Strain Chol1 with the Steroid Compound Cholate Indicates an Aldolytic Reaction Step for Deacetylation of the Side Chain

The Essential Function of Genes for a Hydratase and an Aldehyde Dehydrogenase for Growth of Pseudomonas sp. Strain Chol1 with the Steroid Compound Cholate Indicates an Aldolytic Reaction Step for Deacetylation of the Side Chain

Two Transporters Essential for Reassimilation of Novel Cholate Metabolites by Rhodococcus jostii RHA1

Enzymatic site-selectivity enabled by structure-guided directed evolution

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