Combining Metabolic Engineering and Lipid Droplet Assembly to Achieve Campesterol Overproduction in Saccharomyces cerevisiae.
J Agric Food Chem
; 72(9): 4814-4824, 2024 Mar 06.
Article
in En
| MEDLINE
| ID: mdl-38389392
ABSTRACT
Campesterol is a kind of important functional food additive. Therefore, stable and efficient campesterol biosynthesis is significant. Herein, we first knocked out the sterol 22-desaturase gene in Saccharomyces cerevisiae and expressed sterol Δ7-reductase from Pangasianodon hypophthalmus, obtaining a strain that produced 6.6 mg/L campesterol. Then, the modular expression of campesterol synthesis enzymes was performed, and a campesterol titer of 88.3 mg/L was achieved. Because campesterol is a lipid-soluble macromolecule, we promoted lipid droplet formation by exploring regulatory factors, and campesterol production was improved to 169.20 mg/L. Next, triacylglycerol lipase was used to achieve compartment campesterol synthesis. After enhancing the expression of sterol Δ7-reductase and screening cations, the campesterol titer reached 438.28 mg/L in a shake flask and 1.44 g/L in a 5 L bioreactor, which represents the highest campesterol titer reported to date. Metabolic regulation combined with lipid droplet engineering may be useful for the synthesis of other steroids as well.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Phytosterols
/
Saccharomyces cerevisiae
/
Cholesterol
/
Saccharomyces cerevisiae Proteins
Language:
En
Journal:
J Agric Food Chem
Year:
2024
Type:
Article
Affiliation country:
China