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Total biosynthesis of hydrocortisone from a simple carbon source in yeast

Nature Biotechnology volume 21, pages 143–149 (2003)Cite this article

Abstract

We report on the production of hydrocortisone, the major adrenal glucocorticoid of mammals and an important intermediate of steroidal drug synthesis, from a simple carbon source by recombinant Saccharomyces cerevisiae strains. An artificial and fully self-sufficient biosynthetic pathway involving 13 engineered genes was assembled and expressed in a single yeast strain. Endogenous sterol biosynthesis was rerouted to produce compatible sterols to serve as substrates for the heterologous part of the pathway. Biosynthesis involves eight mammalian proteins (mature forms of CYP11A1, adrenodoxin (ADX), and adrenodoxin reductase (ADR); mitochondrial forms of ADX and CYP11B1; 3β-HSD, CYP17A1, and CYP21A1). Optimization involved modulating the two mitochondrial systems and disrupting of unwanted side reactions associated with ATF2, GCY1, and YPR1 gene products. Hydrocortisone was the major steroid produced. This work demonstrates the feasibility of transfering a complex biosynthetic pathway from higher eukaryotes into microorganisms.

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Figure 1: Reconstitution of the hydrocortisone biosynthetic pathway in S. cerevisiae.
Figure 2: Steroid productivity of selected strains.

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Acknowledgements

This work was funded by Aventis Pharma France. We are in debt to Jacques Raynaud and the late Jean Pierre Lecocq for starting this innovative project, and to Udo Hedtmann for his continuous support. We also thank Francis Karst and Mike Waterman for providing us with their advice during the many years of this exciting adventure.

Author information

Author notes

  1. Gilles Cauet

    Present address: Transgene S.A., 11 Rue de Molsheim, F-67082, Strasbourg, France

  2. David Balbuena & Roberto Spagnoli

    Present address: Aventis Pharma, Lead Discovery Technologies, 102 Route de Noisy, Romainville, F-93235, France

  3. Bruno Dumas

    Present address: Aventis Pharma, Functional Genomics, 13 Quai J. Guesdes, Vitry sur Seine, F-93400, Cedex, France

  4. Florence Ménard Szczebara

    Present address: Anjou Recherche/Vivendi Water, 1 Place de Turenne, 94417, St Maurice Cedex, France

  5. Cathy Chandelier & Stéphane Bourot

    Present address: Bayer Crop Science, 1 rue Pierre Fontaine, F-91058, Evry, France

  6. Catherine Duport

    Present address: UMR408, INRA, F-84914, Avignon, France

  7. Sophie Blanchard

    Present address: Laboratoire écologie moléculaire IBEAS–Université de PAU BP, 1155–64013, Pau, France

  8. Agnès Groisillier

    Present address: Station Biologique de Roscoff, UMR 7127, CNRS, Place Georges Teissier BP 74, 29682, Roscoff Cedex, France

  9. Eric Degryse

    Present address: Centre de Recherche Pernod-Ricard, 120 Avenue du Marechal Foch, 94015, Créteil Cedex, France

  10. Jacques Winter

    Present address: Infors Sarl, 6 Rue Marcel Paul, 91742, Massy Cedex, France

  11. Tilman Achstetter

    Present address: Hochschule Bremen, ISTAB, Neustadtswall 30, D-28201, Bremen, Germany

  12. Florence Ménard Szczebara and Cathy Chandelier: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratoire d'Ingénierie des Protéines Membranaires, CGM du CNRS, Gif sur Yvette, F-91198, France

    Florence Ménard Szczebara, Cathy Chandelier, Coralie Villeret, Amélie Masurel, Stéphane Bourot, Catherine Duport & Denis Pompon

  2. École Supérieure de Technologie des Biomolécules de Bordeaux 2 (ESTBB), Université Victor Segalen Bordeaux 2, 146 Rue Léo Saignat, Bordeaux, F-33076, France

    Sophie Blanchard, Agnès Groisillier, Eric Testet, Patricia Costaglioli & Tilman Achstetter

  3. Transgene S.A., 11 Rue de Molsheim, F-67082, Strasbourg, France

    Eric Degryse & Tilman Achstetter

  4. Aventis Pharma, Lead Discovery Technologies, 102 Route de Noisy, Romainville, F-93235, France

    Jacques Winter

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  1. Florence Ménard Szczebara
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Correspondence to Bruno Dumas.

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Szczebara, F., Chandelier, C., Villeret, C. et al. Total biosynthesis of hydrocortisone from a simple carbon source in yeast. Nat Biotechnol 21, 143–149 (2003). https://doi.org/10.1038/nbt775

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