Self-sufficient biosynthesis of pregnenolone and progesterone in engineered yeast

@article{Duport1998SelfsufficientBO,
  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},
  year={1998},
  volume={16},
  pages={186-189}
}
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… 
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References

SHOWING 1-10 OF 26 REFERENCES
Physiological implications of sterol biosynthesis in yeast.
TLDR
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 (*)
TLDR
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.
Cloning and characterization of the Saccharomyces cerevisiae C-22 sterol desaturase gene, encoding a second cytochrome P-450 involved in ergosterol biosynthesis.
The mitochondrial environment is required for activity of the cholesterol side-chain cleavage enzyme, cytochrome P450scc.
TLDR
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.
Incorporation of steroidogenic pathways which produce cortisol and aldosterone from cholesterol into nonsteroidogenic cells
[Import of a modified form of a cytochrome P450scc precursor into mitochondria from various sources].
TLDR
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.
Advances in microbial steroid biotransformation
Yeast expression of animal and plant P450s in optimized redox environments.
Yeast sterols: yeast mutants as tools for the study of sterol metabolism.
Expression of bovine cytochrome P450c17 cDNA in Saccharomyces cerevisiae.
TLDR
Results indicated that bovine P450c17 synthesized in S. cerevisiae cells manifests the 17 alpha-hydroxylase activity, but not the C17,20-lyase activity.
...
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