Triterpenoids are a class of bioactive secondary metabolites with high added value, widely used in the pharmaceutical and healthcare industries, and lanosterol is the main skeleton for the biosynthesis of triterpenoids. To construct the lanosterol biosynthetic pathway in Escherichia coli chassis, squalene synthase (SQS) derived from Saccharomyces cerevisiae, squalene epoxidase (SE) and oxidosqualene cyclase (OSC) derived from Methylococcus capsulatus were cloned into vectors pET28a (+) and pCDF-DUET-1 respectively, and co-transformed into E. coli BL21 (DE3). The engineered strain BT-tSSE-DXOSC produced 0.77 mg/L lanosterol. Precursor supply was improved by overexpressing key enzymes in the MEP pathway, 1-deoxy-D-xylulose-5-phosphate synthase (DXS) and farnesyl pyrophosphate synthase (FPS), thus lanosterol production reached 2.39 mg/L, increasing by 2.1 times compared to that before optimization. The optimal fermentation conditions were determined by single-factor and orthogonal experiments: pH 7.5, isopropyl-b-D-thiogalactoside (IPTG) concentration 0.5 mmol/L, and inducing temperature 30 ℃, under which the lanosterol production reached 5.12 mg/L, increasing by 1.1 times than that under initial conditions. This study constructed a lanosterol-producing strain for the first time, which provides a platform for the synthesis of triterpenoids and steroids as well as the study of relating metabolic pathways.
HU Miaomiao
,
LU Zhenming
,
LUO Zhishan
,
LI Xinyang
,
XU Guoqiang
,
LI Hui
,
SHI Jinsong
,
XU Zhenghong
. Construction and optimization of lanosterol synthetic pathway in Escherichia coli[J]. Food and Fermentation Industries, 2023
, 49(3)
: 16
-21
.
DOI: 10.13995/j.cnki.11-1802/ts.031643
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