title: De novo biosynthesis of Gastrodin in Escherichia coli.
abstract: 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 545mgl(-1) gastrodin in 48h. This work creates a new route to produce gastrodin, instead of plant extractions and chemical synthesis.
authors.affiliation: Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China; Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China; University of Chinese Academy of Sciences, Beijing, China.
authors.forename: Yanfen
authors.initials: Y
authors.lastname: Bai
authors.affiliation: Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China; Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.
authors.forename: Hua
authors.initials: H
authors.lastname: Yin
authors.affiliation: Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China; Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.
authors.forename: Huiping
authors.initials: H
authors.lastname: Bi
authors.affiliation: Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China; Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.
authors.forename: Yibin
authors.initials: Y
authors.lastname: Zhuang
authors.affiliation: Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China; Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China. Electronic address: liu_t@tib.cas.cn.
authors.forename: Tao
authors.initials: T
authors.lastname: Liu
authors.affiliation: Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.
authors.forename: Yanhe
authors.initials: Y
authors.lastname: Ma
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D001592:Benzyl Alcohols; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005960:Glucosides / Q000096:biosynthesis*; D016695:Glycosyltransferases / Q000096:biosynthesis / Q000235:genetics; D009615:Nocardia / Q000201:enzymology / Q000235:genetics; D010088:Oxidoreductases / Q000096:biosynthesis / Q000235:genetics; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics; D029771:Rhodiola / Q000201:enzymology / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D001592:Benzyl Alcohols; D005960:Glucosides; D010940:Plant Proteins; C045345:gastrodin; D010088:Oxidoreductases; C019825:carboxylic acid reductase; D016695:Glycosyltransferases
keywords: 4-Formylphenyl β-D-glucopyranoside (PubChem CID: 7573800); 4-Hydroxybenzoic acid (PubChem CID: 135); 4-Hydroxybenzoic acid β-D-glucopyranoside (PubChem CID: 440186); 4-Hydroxybenzyl alcohol (PubChem CID: 125); 4-Methylumbelliferone (PubChem CID: 5280567); 4-Methylumbelliferyl β-D-glucoside (PubChem CID: 2733779); Chorismate; Directed evolution; Escherichia coli; Gastrodin; Gastrodin (PubChem CID: 115067); Glycosyltransferase; Metabolic engineering; Salidroside (PubChem CID: 159278); Tyrosol (PubChem CID: 10393)