Expression of rat liver vitamin D3 25-hydroxylase cDNA in Saccharomyces cerevisiae
| Autor: | Emiko Usui, Megumi Akiyoshi-Shibata, Hideo Ohkawa, Toshiyuki Sakaki, Kyuichiro Okuda, Mitsuhidel Noshiro, Yoshiyasu Yabusaki |
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| Rok vydání: | 1991 |
| Předmět: |
Heterologous expression in yeast
Saccharomyces cerevisiae Molecular Sequence Data Restriction Mapping Biophysics Gene Expression Cytochrome P450 Reductase Biochemistry DNA Mitochondrial Transformation Genetic Cytochrome P-450 Enzyme System Structural Biology Complementary DNA Adrenodoxin Gene expression Genetics Animals 5β-Cholestane-3α 7α 12α-triol 27-hydroxylase Promoter Regions Genetic Vitamin D3 25-hydroxylase Molecular Biology chemistry.chemical_classification biology Base Sequence Alcohol Dehydrogenase Cell Biology biology.organism_classification Molecular biology Yeast Mitochondria Rats Enzyme chemistry Liver Steroid Hydroxylases biology.protein Cholestanetriol 26-Monooxygenase |
| Zdroj: | FEBS letters. 280(2) |
| ISSN: | 0014-5793 |
| Popis: | The cDNA coding for the precursor protein of rat liver mitochondrial vitamin D3 25-hydroxylase, cytochrome P450LMT25, was expressed under the control of the yeast alcohol dehydrogenase I promoter and terminator in Saccharomyces cerevisiae AH22 cells. The transformed yeast cells produced a P450LMT25 protein with an almost similar apparent molecular weight as compared with that of the authentic mature enzyme. The expression level of the P450LMT25 hemoprotein was about 5 x 10(4) molecules per cell as determined by reduced CO-difference spectra. The mitochondrial fraction prepared from the transformed yeast cells exhibited both 25-hydroxylase activity toward 1 alpha-hydroxyvitamin D3 and 27-hydroxylase activity toward 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha-triol in a reconstituted system containing bovine adrenodoxin and NADPH-adrenodoxin reductase. |
| Databáze: | OpenAIRE |
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