Engineering a mevalonate pathway in Escherichia coli for production of terpenoids

Abstract

Isoprenoids are the most numerous and structurally diverse family of natural products. Terpenoids, a class of isoprenoids often isolated from plants, are used as commercial flavor and fragrance compounds and antimalarial or anticancer drugs. Because plant tissue extractions typically yield low terpenoid concentrations, we sought an alternative method to produce high-value terpenoid compounds, such as the antimalarial drug artemisinin, in a microbial host. We engineered the expression of a synthetic amorpha-4,11-diene synthase gene and the mevalonate isoprenoid pathway from Saccharomyces cerevisiae in Escherichia coli. Concentrations of amorphadiene, the sesquiterpene olefin precursor to artemisinin, reached 24 microg caryophyllene equivalent/ml. Because isopentenyl and dimethylallyl pyrophosphates are the universal precursors to all isoprenoids, the strains developed in this study can serve as platform hosts for the production of any terpenoid compound for which a terpene synthase gene is available.

Biofuel alternatives to ethanol: pumping the microbial well

“…High-level production of IPP has been achieved in both E. coli and S. cerevisiae, and many different iso-prenoids have been produced with the use of these engin-eered hosts [49][50][51][52][53]. In the engineering of E. coli hosts, the existence of two separate biosynthetic routes has been exploited by introducing the non-native mevalonate path-way.…”

Functional Expression of an Orchid Fragrance Gene in Lactococcus lactis

“…This can be carried out by utilization of the host’s precursors such as geranyl pyrophosphate (GPP) and farnesyl pyrophosphate (FPP) for the production of mono- and sesquiterpenes respectively, provided that the pool of precursors is sufficient to maintain the requirements of both the endogenous pathways as well as the fabricated one. Therefore, efficient production hosts such as Escherichia coli [8,9] and yeasts [10–12] have been metabolically engineered for increased heterologous isoprenoids production.…”

A platform pathway for production of 3-hydroxyacids provides a biosynthetic route to 3-hydroxy-γ-butyrolactone

“…These low titres were observed in spite of the availability of natural pathways to produce the target molecules. The latter example is especially encouraging given more recent reports of 27.4 g l À 1 amorphadiene produced in 2-L fed-batch reactors 52 . Indeed, we anticipate that production of (S)-3,4-DHBA/3-HBL at comparable titres can result in a substantial reduction in cost of goods for high-value applications such as pharmaceutical synthesis.…”