1.Construction and optimization of squalene epoxide synthetic pathway in Escherichia coli.
Xixi SHAO ; Yunhe MENG ; Shenting ZHOU ; Xinting LIU ; Huishu JIANG ; Xiao XIAO ; Jing YANG ; Gongyuan WEI ; Chonglong WANG
Chinese Journal of Biotechnology 2021;37(6):2105-2115
Triterpenoids are a class of natural products of great commercial value that are widely used in pharmaceutical, health care and cosmetic industries. The biosynthesis of triterpenoids relies on the efficient synthesis of squalene epoxide, which is synthesized from the NADPH dependent oxidation of squalene catalyzed by squalene epoxidase. We screened squalene epoxidases derived from different species, and found the truncated squalene epoxidase from Rattus norvegicus (RnSETC) showed the highest activity in engineered Escherichia coli. Further examination of the effect of endogenous cytochrome P450 reductase like (CPRL) proteins showed that overexpression of NADH: quinone oxidoreductase (WrbA) under Lac promoter in a medium-copy number plasmid increased the production of squalene epoxide by nearly 2.5 folds. These results demonstrated that the constructed pathway led to the production of squalene epoxide, an important precursor for the biosynthesis of triterpenoids.
Animals
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Escherichia coli/genetics*
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NADPH-Ferrihemoprotein Reductase
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Protein Engineering
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Rats
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Repressor Proteins
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Squalene
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Squalene Monooxygenase/genetics*