Self-sufficient biosynthesis of pregnenolone and progesterone in engineered yeast

  title={Self-sufficient biosynthesis of pregnenolone and progesterone in engineered yeast},
  author={Catherine Duport and Roberto Spagnoli and Eric Degryse and Denis Pompon},
  journal={Nature Biotechnology},
The first two steps of the steroidogenic pathway were reproduced in Saccharomyces cerevisiae. Engineering of sterol biosynthesis by disruption of the Δ22-desaturase gene and introduction of the Arabidopsis thaliana Δ7-reductase activity and coexpression of bovine side chain cleavage cytochrome P450, adrenodoxin, and adrenodoxin reductase, lead to pregnenolone biosynthesis from a simple carbon source. Following additional coexpression of human β-hydroxysteroid dehydrogenase/isomerase… 
Total biosynthesis of hydrocortisone from a simple carbon source in yeast
This work demonstrates the feasibility of transfering a complex biosynthetic pathway from higher eukaryotes into microorganisms.
From structure and functions of steroidogenic enzymes to new technologies of gene engineering
This review summarizes data about structural and functional organization of steroidogenic P450-dependent enzymatic systems and principal steps in the creation and catalytic properties of transgenic strains of Escherichia coli, Saccharomyces cerevisiae, and Yarrowia lipolytica.
Metabolic engineering strategies for de novo biosynthesis of sterols and steroids in yeast
In this review, the metabolic engineering strategies developed and employed for improving the de novo biosynthesis of sterols and steroids in yeast based on the regulation mechanisms are summarized, and the recent progresses in de noovo synthesis of some typical sterol and steroid compounds in yeast are introduced.
The Fascinating World of Steroids: S. cerevisiae as a Model Organism for the Study of Hydrocortisone Biosynthesis
The purpose of this review is to introduce the reader to the steroid world through an example of Inetabolic engineering consisting of the transfer of a InamlnaJian biosynthetic pathway into a
Escherichia coli kduD encodes an oxidoreductase that converts both sugar and steroid substrates
KduD from E. coli is therefore a promiscuous enzyme that has a predicted role in sugar conversion in vivo but can be used for the production of valuable bioactive 20-hydroxysteroids.
Effect of steroid biosynthesis modifiers on progesterone biotransformation by recombinant yeasts expressing cytochrome P450cl7
Ketokonazole, mifepriston and danazol were found to be low-affinity competitive inhibitors, but the 20-dihydroderivatives of progesterone were mixed type inhibitors of the cytochrome P450cl7.
Biotransformation of steroids by the fission yeast Schizosaccharomyces pombe
The findings indicate that genes encoding soluble proteins should be examined as candidates for actual steroid dehydrogenase activity in Schizosaccharomyces pombe, and more than one steroid dehydration homologue is likely to occur.
Precursor Balancing for Metabolic Engineering of Lycopene Production in Escherichia coli
It is shown that the distribution between pyruvate and glyceraldehyde 3‐phosphate (G3P) is a major factor that can limit isoprenoid production yields in E. coli, and modifications that achieve a more equitable distribution between the two precursors are able to increase the lycopene yield in metabolically engineered E. bacteria.


Physiological implications of sterol biosynthesis in yeast.
Questions why fungi require ergosterol rather than the simpler cholesterol provides heuristic impetus for further experimentation and research on the effects of altering sterol metabolism by sterol mutants are providing new insights into sterol functions in the cells.
Cloning by Metabolic Interference in Yeast and Enzymatic Characterization of Arabidopsis thaliana Sterol 7-Reductase (*)
In vitro tests indicated that Δ7-reductase activity is preferentially associated with the endoplasmic reticulum membrane and confirmed the previous finding that NADPH is the reducing agent, and analysis using transformed wild type yeast or sterol altered mutants suggested that Δ5,7-ergosta- and cholesta-sterols are efficiently reduced in vivo, regardless of the structural variations on the side chain.
The mitochondrial environment is required for activity of the cholesterol side-chain cleavage enzyme, cytochrome P450scc.
Results show that P450scc can function only in the mitochondria, and it appears to be the mitochondrial environment that is required, rather than the specific mitochondrial electron-transport intermediates.
[Import of a modified form of a cytochrome P450scc precursor into mitochondria from various sources].
An Escherichia coli strain providing hypersynthesis of a recombinant cytochrome P450scc precursor supplemented with the extra MetArgGlySerHis6GlyIleArg sequence at the NH2-terminus has been constructed and it has been found that the recombinant precursor is imported into isolated rat liver and rat heart mitochondria as well as into yeast mitochondria.
Expression of bovine cytochrome P450c17 cDNA in Saccharomyces cerevisiae.
Results indicated that bovine P450c17 synthesized in S. cerevisiae cells manifests the 17 alpha-hydroxylase activity, but not the C17,20-lyase activity.