title: Total biosynthesis of hydrocortisone from a simple carbon source in yeast.
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; 3beta-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.
authors.affiliation: Laboratoire d'Ingénierie des Protéines Membranaires, CGM du CNRS, F-91198 Gif sur Yvette, France.
authors.forename: Florence Ménard
authors.initials: FM
authors.lastname: Szczebara
authors.forename: Cathy
authors.initials: C
authors.lastname: Chandelier
authors.forename: Coralie
authors.initials: C
authors.lastname: Villeret
authors.forename: Amélie
authors.initials: A
authors.lastname: Masurel
authors.forename: Stéphane
authors.initials: S
authors.lastname: Bourot
authors.forename: Catherine
authors.initials: C
authors.lastname: Duport
authors.forename: Sophie
authors.initials: S
authors.lastname: Blanchard
authors.forename: Agnès
authors.initials: A
authors.lastname: Groisillier
authors.forename: Eric
authors.initials: E
authors.lastname: Testet
authors.forename: Patricia
authors.initials: P
authors.lastname: Costaglioli
authors.forename: Gilles
authors.initials: G
authors.lastname: Cauet
authors.forename: Eric
authors.initials: E
authors.lastname: Degryse
authors.forename: David
authors.initials: D
authors.lastname: Balbuena
authors.forename: Jacques
authors.initials: J
authors.lastname: Winter
authors.forename: Tilman
authors.initials: T
authors.lastname: Achstetter
authors.forename: Roberto
authors.initials: R
authors.lastname: Spagnoli
authors.forename: Denis
authors.initials: D
authors.lastname: Pompon
authors.forename: Bruno
authors.initials: B
authors.lastname: Dumas
mesh_terms: D000818:Animals; D002244:Carbon / Q000378:metabolism*; D002417:Cattle; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism; D000431:Ethanol / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D005818:Genetic Engineering / Q000379:methods*; D005947:Glucose / Q000378:metabolism; D006801:Humans; D006854:Hydrocortisone / Q000096:biosynthesis* / Q000235:genetics; D009097:Multienzyme Complexes / Q000235:genetics / Q000378:metabolism; D011786:Quality Control; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D013045:Species Specificity
chemical_list: D009097:Multienzyme Complexes; D011994:Recombinant Proteins; D000431:Ethanol; D002244:Carbon; D003577:Cytochrome P-450 Enzyme System; D005947:Glucose; D006854:Hydrocortisone