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US 7,977,065 B2Granted
Yeast strains autonomously producing steroids
AssigneeAventis Pharma S.A.
InventorsRoberto Spagnoli, Tilman Achstetter, Gilles Cauet, Eric Degryse, Bruno Dumas, Denis Pompon, Jacques Winter
Life of this patent
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Abstract
The present invention relates to genetically modified yeast strains autonomously producing, from a simple carbon source, steroids. The invention also relates to a method for producing steroids from such yeast strains.
Claims22
A method of producing a steroid or a steroid derivative, comprising
- a) fermenting a Saccharomyces cerevisiae yeast strain genetically modified to autonomously produce, from a simple carbon source, a steroid or steroid derivative derived from cholesterol metabolism, wherein said steroid or steroid derivative is selected from the group consisting of pregnenolone, 17α-hydroxypregnenolone, cortisol, cortexolone, progesterone, 17α-hydroxyprogesterone, and derivatives thereof;
- wherein the genetic modification comprises i) the inactivation of ATF2, GCY1 and YPR1, and ii) at least one heterologous gene or cDNA in an expression cassette, wherein the gene or cDNA is selected from the group consisting of the gene or cDNA encoding A. thaliana sterol Δ7-reductase, bovine cytochrome P450SCC, bovine adrenodoxin, bovine adremodoxin reductase, bovine 3β-hydroxysteroid dehydrogenase/isomerase, human 3β-hydroxysteroid dehydrogenase/isomerase, bovine cytochrome P450 C17, human cytochrome P450 C21, human cytochrome P450 C11, bovine cytochrome P450 C11 and human-bovine hybrid cytochrome P450 C11, and
- b) recovering the steroid or the steroid derivative produced.
A method of producing a steroid or a steroid derivative, comprising
- a) fermenting a Saccharomyces cerevisiae yeast strain genetically modified to autonomously produce, from a simple carbon source, a steroid or steroid derivative derived from cholesterol metabolism, wherein said steroid or steroid derivative is selected from the group consisting of pregnenolone, 17α-hydroxypregnenolone, cortisol, cortexolone, progesterone, 17α-hydroxyprogesterone, and derivatives thereof;
- wherein the genetic modification comprises i) the inactivation of ADE2, ATF2, GCY1 and YPR1, and ii) at least one heterologous gene or cDNA in an expression cassette, wherein the gene or cDNA is selected from the group consisting of the gene or cDNA encoding A. thaliana sterol Δ7-reductase, bovine cytochrome P450SCC, bovine adrenodoxin, bovine adrenodoxin reductase, bovine 3β-hydroxysteroid dehydrogenase/isomerase, human 3β-hydroxysteroid dehydrogenase/isomerase, bovine cytochrome P450 C17, human cytochrome P450 C21, human cytochrome P450 C11, bovine cytochrome P450 C11 and human-bovine hybrid cytochrome P450 C11, and
- b) recovering the steroid or the steroid derivative produced.
A method of producing a steroid or a steroid derivative, comprising
- a) fermenting a Saccharomyces cerevisiae yeast strain genetically modified to autonomously produce, from a simple carbon source, a steroid or steroid derivative derived from cholesterol metabolism, wherein said steroid or steroid derivative is selected from the group consisting of pregnenolone, 17α-hydroxypregnenolone, cortisol, cortexolone, progesterone, 17α-hydroxyprogesterone, and derivatives thereof;
- wherein the genetic modification comprises i) the inactivation of ERG5, ATF2, GCY1 and YPR1, and ii) at least one heterologous gene or cDNA in an expression cassette, wherein the gene or cDNA is selected from the group consisting of the gene or cDNA encoding A. thaliana sterol Δ7-reductase, bovine cytochrome P450SCC, bovine adrenodoxin, bovine adrenodoxin reductase, bovine 3β-hydroxysteroid dehydrogenase/isomerase, human 3β-hydroxysteroid dehydrogenase/isomerase, bovine cytochrome P450 C17, human cytochrome P450 C21, human cytochrome P450 C11, bovine cytochrome P450 C11 and human-bovine hybrid cytochrome P450 C11, and
- b) recovering the steroid or the steroid derivative produced.
A method of producing a steroid or a steroid derivative, comprising
- a) a Saccharomyces cerevisiae yeast strain genetically modified to autonomously produce, from a simple carbon source, a steroid or steroid derivative derived from cholesterol metabolism, wherein said steroid or steroid derivative is selected from the group consisting of pregnenolone, 17α-hydroxypregnenolone, cortisol, cortexolone, progesterone, 17α-hydroxyprogesterone, and derivatives thereof;
- wherein the genetic modification comprises i) the inactivation of ADE2, ERG5, ATF2, GCY1 and YPR1, and ii) at least one heterologous gene or cDNA in an expression cassette, wherein the gene or cDNA is selected from the group consisting of the gene or cDNA encoding A. thaliana sterol Δ7-reductase, bovine cytochrome P450SCC, bovine adrenodoxin, bovine adrenodoxin reductase, bovine 3β-hydroxysteroid dehydrogenase/isomerase, human 3β-hydroxysteroid dehydrogenase/isomerase, bovine cytochrome P450 C17, human cytochrome P450 C21, human cytochrome P450 C11, bovine cytochrome P450 C11 and human-bovine hybrid cytochrome P450 C11, and
- b) recovering the steroid or the steroid derivative produced.
Dependencies are inferred from the claim text.