Hydrocortisone production using whole-cell biocatalysts in recombinant Escherichia coli

Hydrocortisone, is one of the most powerful adrenocorticosteroid used in clinics for glucose metabolism. It is also served as an important intermediate for synthesis of other steroid drugs. However, hydrocortisone is currently made by multiple chemical steps. All of these chemical routes for the synthesis of hydrocortisone suffered from certain defects. With continuous development of biotechnology, the prospect of biosynthesis of hydrocortisone drugs is unanimously favored. In this paper, we reported that hydrocortisone was made for the first time from progesterone using engineered Escherichia coli strains with assembled P450s in one pot artificial cascade reactions. This biocatalytic cascade comprises three different P450 modules named 17AM, 21 M and 11BM. Firstly, an efficient biosynthetic pathway of deoxycortisone from progesterone was constructed by assembling 17AM and 21 M modules. The results revealed that the catalytic efficiency of the strain deo04(pRSF-21M2–17AM) is significantly higher (86.74 μmol·L−1) than the other combinatorial strains. Subsequently, the three-P450-modules co-expression system of E. coli C43(DE3) named hyg4 was developed to produce hydrocortisone directly from progesterone. Moreover, the whole-cell biotransformation system was optimized for culture and reaction conditions. The total production of hydrocortisone reached 16.82 μmol·L−1 within 24 h. •Hydrocortisone was synthesized for the first time from progesterone using E.coli.•Deoxycortisol was efficiently produced by expressing 17AM and 21 M in one plasmid.•Two-fold expression of Fpr and Adx enhanced hydrocortisone biosynthesis.•A cascade reaction consisting of three P450s was first constructed in E.coli.•Provide a strategy for the cell factory to produce polyhydroxylated products.