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Purification and reconstitution of activity of Saccharomyces cerevisiae P450 61, a sterol Δ 22 ‐desaturase
Author(s) -
Kelly Steven L.,
Lamb David C.,
Corran Andrew J.,
Baldwin Brian C.,
Parks Leo W.,
Kelly Diane E.
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)01342-3
Subject(s) - saccharomyces cerevisiae , sterol , ergosterol , biochemistry , microsome , enzyme , biology , yeast , reductase , biosynthesis , chemistry , microbiology and biotechnology , cholesterol
P450 was purified from microsomal fractions of a strain of Saccharomyces cerevisiae which contained detectable P450 despite the disruption of CYP51A1 . The P450 had a molecular mass of 58 kDa, similar to P450 51A1, and in a reconstituted assay with rabbit NADPH‐P450 reductase and dilauryl phosphotidylcholine exhibited activity for conversion of ergosta‐5,7‐dienol into ergosterol. N‐Terminal amino acid sequencing of the purified protein corresponded to the translated sequence of P450 61 which was recently identified during sequencing of chromosome XIII. This allowed the function of this family of P450 to be identified as sterol Δ 22 ‐desaturation in the pathway of ergosterol biosynthesis.