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4-Methylzymosterone and Other Intermediates of Sterol Biosynthesis from Yeast Mutants Engineered in the ERG27 Gene Encoding 3-Ketosteroid Reductase

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Lipids

Abstract

Studies in the post-squalene section of sterol biosynthesis may be hampered by the poor availability of authentic standards. The present study used different yeast strains engineered in 3-ketosteroid reductase (Erg27p) to obtain radioactive and non-radioactive intermediates of sterol biosynthesis hardly or not available commercially. Non-radioactive 3-keto 4-monomethyl sterones were purified from non-saponifiable lipids extracted from cells bearing point-mutated 3-ketosteroid reductase. Two strategies were adopted to prepare the radioactive compounds: (1) incubation of cell homogenates of an ERG27-deletant strain with radioactive lanosterol, (2) incubation of growing cells of a strain expressing point-mutated 3-ketosteroid reductase with radioactive acetate. Chemical reduction of both radioactive and non-radioactive 3-keto sterones gave the physiological 3-β OH sterols, as well as the non-physiological 3-α OH isomers. This combined biological and chemical preparation procedure provided otherwise unavailable or hardly available 4-mono-methyl intermediates of sterol biosynthesis, paving the way for research into their roles in physiological and pathological conditions.

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Abbreviations

OSC/Erg7p:

Yeast oxidosqualene cyclase

ERG7 :

Gene encoding oxidosqualene cyclase

Erg27p:

Yeast 3-ketosteroid reductase

ERG27 :

Gene encoding 3-ketosteroid reductase

HSD17B7:

Hydroxysteroid-17β-dehydrogenase type 7

SC4MOL:

Sterol-C-4 methyloxidase-like

NSDHL:

NADH sterol dehydrogenase-like

C14ORF1:

Chromosome 14 Open Reading Frame 1

MAS:

Meiosis-activating sterols

EGFR:

Epidermal growth factor receptor

SAM:

S-adenosyl methionine

TLC:

Thin-layer chromatography

CI–MS:

Chemical ionization-mass spectrometry

GC–MS:

Gas chromatography-mass spectrometry

NMR:

Nuclear magnetic resonance

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Acknowledgments

We thank M. Bard for providing some of the yeast strains used in this work. We also thank F. Viola for helpful discussions and revision of the manuscript.This research was supported by the financial support of the University of Torino [Fondo di Ricerca Locale (ex 60 %) 2013—linea B] to Simonetta Oliaro-Bosso.

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Authors and Affiliations

  1. Department of Science and Drug Technology, University of Torino, Via P. Giuria 9, 10125, Turin, Italy

    Terenzio Ferrante, Alessandro Barge, Silvia Taramino, Simonetta Oliaro-Bosso & Gianni Balliano

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  1. Terenzio Ferrante
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  2. Alessandro Barge
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  3. Silvia Taramino
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  4. Simonetta Oliaro-Bosso
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  5. Gianni Balliano
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Correspondence to Simonetta Oliaro-Bosso or Gianni Balliano.

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The authors declare no financial or commercial conflict of interest.

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Ferrante, T., Barge, A., Taramino, S. et al. 4-Methylzymosterone and Other Intermediates of Sterol Biosynthesis from Yeast Mutants Engineered in the ERG27 Gene Encoding 3-Ketosteroid Reductase. Lipids 51, 1103–1113 (2016). https://doi.org/10.1007/s11745-016-4173-6

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  • DOI: https://doi.org/10.1007/s11745-016-4173-6

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