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4‐Methylzymosterone and Other Intermediates of Sterol Biosynthesis from Yeast Mutants Engineered in the ERG27 Gene Encoding 3‐Ketosteroid Reductase
Author(s) -
Ferrante Terenzio,
Barge Alessandro,
Taramino Silvia,
OliaroBosso Simonetta,
Balliano Gianni
Publication year - 2016
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/s11745-016-4173-6
Subject(s) - sterol , biochemistry , reductase , ketosteroid , biosynthesis , lanosterol , yeast , mutant , isomerase , metabolic engineering , biology , chemistry , enzyme , gene , cholesterol
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.