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Pregnenolone Overproduction in Yarrowia lipolytica by Integrative Components Pairing of the Cytochrome P450scc System
- Source :
- ACS Synthetic Biology. 8:2666-2678
- Publication Year :
- 2019
- Publisher :
- American Chemical Society (ACS), 2019.
-
Abstract
- Microbial production of steroid drugs exhibits great potentials in much greener and more sustainable manners, in which engineering multiple cytochrome P450s is the prerequisite requirement. The pairing of multicomponents of P450 systems is a tremendous challenge. Herein, biosynthesis of pregnenolone (a common precursor of steroid drugs) in Yarrowia lipolytica was taken as a typical instance to explore the engineering strategy of the cytochrome P450 side-chain cleavage enzyme (P450scc) system. The mature forms of the components belonging to P450scc system, CYP11A1, adrenodoxin (Adx), and adrenodoxin reductase (AdR), were coexpressed in a former constructed campesterol producing strain. To maximize pregnenolone production, an integrative components pairing strategy was proposed for pairing the component sources and balancing the expression levels of CYP11A1, Adx, and AdR. Led by the above approaches, a 2344-fold improvement of pregnenolone titer was achieved at the shake flask level. Consequently, a highest reported pregnenolone titer of 78.0 mg/L in microbes was obtained in a 5 L bioreactor. Our study not only highlights the integrative components pairing of cytochrome P450scc as a general strategy for engineering other cytochrome P450s, but also provides a feasible and efficient platform of Y. lipolytica for other steroids production.
- Subjects :
- 0106 biological sciences
endocrine system
0303 health sciences
biology
Cytochrome
Chemistry
Cholesterol side-chain cleavage enzyme
Biomedical Engineering
Cytochrome P450
Yarrowia
General Medicine
biology.organism_classification
01 natural sciences
Biochemistry, Genetics and Molecular Biology (miscellaneous)
Adrenodoxin reductase
03 medical and health sciences
Biochemistry
010608 biotechnology
Adrenodoxin
biology.protein
Pregnenolone
medicine
Overproduction
030304 developmental biology
medicine.drug
Subjects
Details
- ISSN :
- 21615063
- Volume :
- 8
- Database :
- OpenAIRE
- Journal :
- ACS Synthetic Biology
- Accession number :
- edsair.doi...........d4c5593eca9ffddb66a65826805c7b6b