title: Production of serotonin by dual expression of tryptophan decarboxylase and tryptamine 5-hydroxylase in Escherichia coli.
abstract: A plant-specific biogenic amine, serotonin, was produced by heterologous expression of two key biosynthetic genes, tryptophan decarboxylase (TDC) and tryptamine 5-hydroxylase (T5H), in Escherichia coli. The native T5H, a cytochrome P450 enzyme, was unable to be functionally expressed in E. coli. Through a series of N-terminal deletions or additions of tagging proteins, we generated a functional T5H enzyme construct (GST∆37T5H) in which glutathione S transferase (GST) was translationally fused with the N-terminal 37 amino acid deleted T5H. Dual expression of GST∆37T5H and TDC using a pCOLADuet-1 E. coli vector produced serotonin at concentrations of approximately 24 mg l⁻¹ in the culture medium and 4 mg l⁻¹ in the cells. An optimum temperature of approximately 20 °C was required to achieve peak serotonin production in E. coli because the low induction temperature gave rise to the highest soluble expression of GST∆37T5H.
authors.affiliation: Department of Biotechnology, Interdisciplinary Programs of Graduate School for Bioenergy and Biomaterials, Bioenergy Research Center, Chonnam National University, Gwangju 500-757, South Korea.
authors.forename: Sangkyu
authors.initials: S
authors.lastname: Park
authors.forename: Kiyoon
authors.initials: K
authors.lastname: Kang
authors.forename: Shin Woo
authors.initials: SW
authors.lastname: Lee
authors.forename: Mi-Jeong
authors.initials: MJ
authors.lastname: Ahn
authors.forename: Jung-Myung
authors.initials: JM
authors.lastname: Bae
authors.forename: Kyoungwhan
authors.initials: K
authors.lastname: Back
mesh_terms: D001142:Aromatic-L-Amino-Acid Decarboxylases / Q000235:genetics / Q000378:metabolism*; D003470:Culture Media / Q000737:chemistry; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D005822:Genetic Vectors; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012701:Serotonin / Q000096:biosynthesis*
chemical_list: D003470:Culture Media; D050505:Mutant Proteins; D011993:Recombinant Fusion Proteins; D012701:Serotonin; D003577:Cytochrome P-450 Enzyme System; C407366:tryptamine 5-hydroxylase; D001142:Aromatic-L-Amino-Acid Decarboxylases