Improved campesterol production in engineered Yarrowia lipolytica strains
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To engineer Yarrowia lipolytica for improving the heterologous production of campesterol (a key precursor to manufacture pharmaceutical steroids).
By screening 7-dehydrocholesterol reductase (DHCR7) from diverse species, DHCR7 from Danio rerio was the best candidate for campesterol synthesis. Overexpression of ACL (ATP: citrate lyase) or POX2 (peroxisome acyl-CoA oxidase 2) were key to improving campesterol production. The highest yield of campesterol was 942 mg/l was with the strain overexpressing POX2 in a 5 l bioreactor via high cell density fermentation process with a restricted supply of carbon sourc, sunflower seed oil.
A promising platform to synthesize downstream steroid drugs was established. Efficient approaches were provided to improve the production of desired molecules in Y. lipolytica with high oil utilization efficiency.
KeywordsCampesterol 7-Dehydrocholesterol reductase Metabolic engineering Oil utilization Synthetic biology Yarrowia lipolytica
This work was supported by the financial support from the International S&T Cooperation Program of China (2015DFA00960), the National Natural Science Foundation of China (21390203, 31570088 and 21622605) and Innovative Talents and Platform Program of Tianjin (16PTSYJC00050).
Supplementary Table 1—Plasmids and strains used in this study.
Supplementary Table 2—Primers used in this study.
Supplementary Table 3—The Codon-optimized sequences of 7-dehydrocholesterol reductase (DHCR7) involved in this study.
Supplementary Fig. 1—Analysis of sterols profile of Y. lipolytica strains SyBE_Yl02060002 and SyBE_Yl02060004–08 for DHCR7 screening.
Supplementary Fig. 2—GC-TOF/MS analysis of campesterol producing strains SyBE_Yl02060006 and SyBE_Yl02060056.
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