Expression of cytochrome P-450d by Saccharomyces cerevisiae

Toru Shimizu; Kazuhiro Sogawa; Yoshiaki Fujii-Kuriyama; Masae Takahashi; Yoshiro Ogoma; Masahiro Hatano

doi:10.1016/0014-5793(86)81491-0

Expression of cytochrome P-450_d by Saccharomyces cerevisiae

Toru Shimizu, Kazuhiro Sogawa, Yoshiaki Fujii-Kuriyama, Masae Takahashi, Yoshiro Ogoma, Masahiro Hatano

Research output: Contribution to journal › Article › peer-review

23 Citations (Scopus)

Abstract

Rat liver microsomal cytochrome P-450_d was abundantly expressed in the yeast Saccharomyces cerevisiae by using a yeast-Escherichia coli shuttle vector consisting of rat liver P-450_d cDNA and yeast acid phosphatase promoter. The expressed cytochrome P-450_d was immunologically crossed with rat liver P-450_d. The hydroxylase activity of estra-1,3,5(10)-triene-3,17β-diol was 11 nmol/min per nmol P-450_d, which is comparable to that reported previously for rat liver P-450_d. The expressed P-450_d content was nearly 1% of total yeast protein as estimated from immunoblotting, hydroxylase activity and optical absorption of the reduced CO form.

Original language	English
Pages (from-to)	217-221
Number of pages	5
Journal	FEBS Letters
Volume	207
Issue number	2
DOIs	https://doi.org/10.1016/0014-5793(86)81491-0
Publication status	Published - 1986 Oct 27

Keywords

(Yeast)
Cytochrome P-450
Expression
cDNA

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

Access to Document

10.1016/0014-5793(86)81491-0

Cite this

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title = "Expression of cytochrome P-450d by Saccharomyces cerevisiae",

abstract = "Rat liver microsomal cytochrome P-450d was abundantly expressed in the yeast Saccharomyces cerevisiae by using a yeast-Escherichia coli shuttle vector consisting of rat liver P-450d cDNA and yeast acid phosphatase promoter. The expressed cytochrome P-450d was immunologically crossed with rat liver P-450d. The hydroxylase activity of estra-1,3,5(10)-triene-3,17β-diol was 11 nmol/min per nmol P-450d, which is comparable to that reported previously for rat liver P-450d. The expressed P-450d content was nearly 1% of total yeast protein as estimated from immunoblotting, hydroxylase activity and optical absorption of the reduced CO form.",

keywords = "(Yeast), Cytochrome P-450, Expression, cDNA",

author = "Toru Shimizu and Kazuhiro Sogawa and Yoshiaki Fujii-Kuriyama and Masae Takahashi and Yoshiro Ogoma and Masahiro Hatano",

note = "Funding Information: T. Kuga for instruction on the transformation of the yeast and to Dr H. Ohkawa for valuable discussions. We gratefully acknowledge Dr K. Matsubara of Osaka University for giving us access to the yeast strain AH22 and the expression vector, pAM82. This work was supported in part by Grants-in-Aid from the Ministry of Education, Science and Culture of Japan.",

year = "1986",

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doi = "10.1016/0014-5793(86)81491-0",

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T1 - Expression of cytochrome P-450d by Saccharomyces cerevisiae

AU - Shimizu, Toru

AU - Sogawa, Kazuhiro

AU - Fujii-Kuriyama, Yoshiaki

AU - Takahashi, Masae

AU - Ogoma, Yoshiro

AU - Hatano, Masahiro

N1 - Funding Information: T. Kuga for instruction on the transformation of the yeast and to Dr H. Ohkawa for valuable discussions. We gratefully acknowledge Dr K. Matsubara of Osaka University for giving us access to the yeast strain AH22 and the expression vector, pAM82. This work was supported in part by Grants-in-Aid from the Ministry of Education, Science and Culture of Japan.

PY - 1986/10/27

Y1 - 1986/10/27

N2 - Rat liver microsomal cytochrome P-450d was abundantly expressed in the yeast Saccharomyces cerevisiae by using a yeast-Escherichia coli shuttle vector consisting of rat liver P-450d cDNA and yeast acid phosphatase promoter. The expressed cytochrome P-450d was immunologically crossed with rat liver P-450d. The hydroxylase activity of estra-1,3,5(10)-triene-3,17β-diol was 11 nmol/min per nmol P-450d, which is comparable to that reported previously for rat liver P-450d. The expressed P-450d content was nearly 1% of total yeast protein as estimated from immunoblotting, hydroxylase activity and optical absorption of the reduced CO form.

AB - Rat liver microsomal cytochrome P-450d was abundantly expressed in the yeast Saccharomyces cerevisiae by using a yeast-Escherichia coli shuttle vector consisting of rat liver P-450d cDNA and yeast acid phosphatase promoter. The expressed cytochrome P-450d was immunologically crossed with rat liver P-450d. The hydroxylase activity of estra-1,3,5(10)-triene-3,17β-diol was 11 nmol/min per nmol P-450d, which is comparable to that reported previously for rat liver P-450d. The expressed P-450d content was nearly 1% of total yeast protein as estimated from immunoblotting, hydroxylase activity and optical absorption of the reduced CO form.

KW - (Yeast)

KW - Cytochrome P-450

KW - Expression

KW - cDNA

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