title: (R,S)-tetrahydropapaveroline production by stepwise fermentation using engineered Escherichia coli.
abstract: Tetrahydropapaveroline (THP), a benzylisoquinoline alkaloid (BIA) found in diverse pharmaceutical compounds, is used as a starting material for the production of BIA. THP also has various neurobiological properties but is difficult to synthesize. Therefore, a simple method for THP production is desired. Recent studies have shown that microbes, especially bacteria, can serve as platforms for synthesizing these complex compounds; however, because bacteria lack organelles, the designed synthetic pathway cannot be compartmentalized. Thus, the metabolic flow is frequently inhibited or disrupted by undesirable reactions. Indeed, in the first attempt to synthesize THP using a single strain of engineered Escherichia coli, the yield was quite low (<5 μM), mainly because of the oxidation of THP by tyrosinase, an essential enzyme in our production system. To circumvent these problems, we constructed a stepwise (R,S)-THP production system, in which the dopamine-producing step and the subsequent THP-producing step were separated. The yield of (R,S)-THP reached 1.0 mM (287 mg/L), the highest yielding BIA production method using a microbe reported to date. Furthermore, we demonstrated that (R,S)-THP produced by stepwise fermentation is useful for the production of reticuline, an important BIAs intermediate. Based on these observations, applying the stepwise fermentation method is discussed.
authors.affiliation: Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi-shi, Ishikawa 921-8836, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Nakagawa
authors.affiliation: Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi-shi, Ishikawa 921-8836, Japan.
authors.forename: Chiaki
authors.initials: C
authors.lastname: Matsuzaki
authors.affiliation: Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi-shi, Ishikawa 921-8836, Japan.
authors.forename: Eitaro
authors.initials: E
authors.lastname: Matsumura
authors.affiliation: Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi-shi, Ishikawa 921-8836, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Koyanagi
authors.affiliation: Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi-shi, Ishikawa 921-8836, Japan.
authors.forename: Takane
authors.initials: T
authors.lastname: Katayama
authors.affiliation: Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi-shi, Ishikawa 921-8836, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Yamamoto
authors.affiliation: Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Oiwake-cho, Kitashirakawa, Sakyo-ku, Kyoto 606-8502, Japan.
authors.forename: Fumihiko
authors.initials: F
authors.lastname: Sato
authors.affiliation: Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi-shi, Ishikawa 921-8836, Japan.
authors.forename: Hidehiko
authors.initials: H
authors.lastname: Kumagai
authors.affiliation: Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi-shi, Ishikawa 921-8836, Japan.
authors.forename: Hiromichi
authors.initials: H
authors.lastname: Minami
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005285:Fermentation; D060847:Metabolic Engineering; D014442:Monophenol Monooxygenase / Q000235:genetics* / Q000378:metabolism; D013765:Tetrahydropapaveroline / Q000138:chemical synthesis* / Q000378:metabolism
chemical_list: D013765:Tetrahydropapaveroline; D014442:Monophenol Monooxygenase