Valeriana officinalis is a medicinal herb which produces a suite of compounds in its root tissue useful for treatment of anxiety and
insomnia. The
sesquiterpene components of the root extract,
valerenic acid and valerena-1,10-diene, are thought to contribute to most of the observed
anxiolytic of
Valerian root preparations. However,
valerenic acid and its biosynthetic intermediates are only produced in low quantities in the roots of V. officinalis. Thus, in this report, Escherichia coli was metabolically engineered to produce substantial quantities of valerena-1,10-diene in shake flask fermentations with
decane overlay. Expression of the wildtype
valerenadiene synthase gene (pZE-wvds) resulted in production of 12μg/mL in LB cultures using endogenous FPP metabolism. Expression of a
codon-optimized version of the
valerenadiene synthase gene (pZE-cvds) resulted in 3-fold higher titers of
valerenadiene (32μg/mL). Co-expression of pZE-cvds with an engineered methyl
erythritol phosphate (MEP) pathway improved
valerenadiene titers 65-fold to 2.09mg/L
valerenadiene. Optimization of the fermentation medium to include
glycerol supplementation enhanced yields by another 5.5-fold (11.0mg/L
valerenadiene). The highest production of
valerenadiene resulted from engineering the
codon-optimized
valerenadiene synthase gene under strong Ptrc and PT7 promoters and via co-expression of an exogenous
mevalonate (MVA) pathway. These efforts resulted in an E. coli production strain that produced 62.0mg/L
valerenadiene (19.4mg/L/OD600 specific productivity). This E. coli production platform will serve as the foundation for the synthesis of novel
valerenic acid analogues potentially useful for the treatment of
anxiety disorders.