De novo biosynthesis of Gastrodin in Escherichia coli

• An artificial novel biosynthetic pathway for gastrodin was constructed.
• Gastrodin was produced from glucose in an E. coli factory for the first time.
• Pathway genes were overexpressed to enhance metabolic flux toward aglycon.
• Directed evolution was applied to improve catalytic properties of UGT73B6.
• ∽545 mg/L of gastrodin was produced by the finally engineered E. coli.

Gastrodin, a phenolic glycoside, is the key ingredient of Gastrodia elata, a notable herbal plant that has been used to treat various conditions in oriental countries for centuries. Gastrodin is extensively used clinically for its sedative, hypnotic, anticonvulsive and neuroprotective properties in China. Gastrodin is usually produced by plant extraction or chemical synthesis, which has many disadvantages. Herein, we report unprecedented microbial synthesis of gastrodin via an artificial pathway. A Nocardia carboxylic acid reductase, endogenous alcohol dehydrogenases and a Rhodiola glycosyltransferase UGT73B6 transformed 4-hydroxybenzoic acid, an intermediate of ubiquinone biosynthesis, into gastrodin in Escherichia coli. Pathway genes were overexpressed to enhance metabolic flux toward precursor 4-hydroxybenzyl alcohol. Furthermore, the catalytic properties of the UGT73B6 toward phenolic alcohols were improved through directed evolution. The finally engineered strain produced 545 mg l−1 gastrodin in 48 h. This work creates a new route to produce gastrodin, instead of plant extractions and chemical synthesis.