Total biosynthesis of hydrocortisone from a simple carbon source in yeast


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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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.

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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).

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