title: Precursor-directed biosynthesis of 5-hydroxytryptophan using metabolically engineered E. coli.
abstract: A novel biosynthetic pathway was designed and verified reversely leading to the production of 5-hydroxytryptophan (5-HTP) from glucose. This pathway takes advantage of the relaxed substrate selectivities of relevant enzymes without employing the unstable tryptophan 5-hydroxylase. First, high-titer of 5-HTP was produced from 5-hydroxyanthranilate (5-HI) by the catalysis of E. coli TrpDCBA. Then, a novel salicylate 5-hydroxylase was used to convert the non-natural substrate anthranilate to 5-HI. After that, the production of 5-HI from glucose was achieved and optimized with modular optimization. In the end, we combined the full pathway and adopted a two-stage strategy to realize the de novo production of 5-HTP. This work demonstrated the application of enzyme promiscuity in non-natural pathway design.
authors.affiliation: †State Key Laboratory of Chemical Resource Engineering, Beijing University of Chemical Technology, Beijing 100029, China.
authors.forename: Xinxiao
authors.initials: X
authors.lastname: Sun
authors.affiliation: ‡College of Engineering, University of Georgia, Athens, Georgia 30602, United States.
authors.forename: Yuheng
authors.initials: Y
authors.lastname: Lin
authors.affiliation: †State Key Laboratory of Chemical Resource Engineering, Beijing University of Chemical Technology, Beijing 100029, China.
authors.forename: Qipeng
authors.initials: Q
authors.lastname: Yuan
authors.affiliation: §BioChemical Engineering Program, College of Engineering, University of Georgia, Athens, Georgia 30602, United States.
authors.forename: Yajun
authors.initials: Y
authors.lastname: Yan
mesh_terms: D006916:5-Hydroxytryptophan / Q000096:biosynthesis* / Q000235:genetics*; D053898:Biosynthetic Pathways / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D005947:Glucose / Q000235:genetics; D060847:Metabolic Engineering / Q000379:methods; D006899:Mixed Function Oxygenases / Q000235:genetics; D014364:Tryptophan / Q000235:genetics
chemical_list: D014364:Tryptophan; D006916:5-Hydroxytryptophan; D006899:Mixed Function Oxygenases; C099614:salicylic acid 5-hydroxylase; D005947:Glucose
keywords: 5-hydroxyanthranilate; 5-hydroxytryptophan; precursor-directed biosynthesis; salicylate 5-hydroxylase