title: Microbial production of plant benzylisoquinoline alkaloids.
abstract: Benzylisoquinoline alkaloids, such as the analgesic compounds morphine and codeine, and the antibacterial agents berberine, palmatine, and magnoflorine, are synthesized from tyrosine in the Papaveraceae, Berberidaceae, Ranunculaceae, Magnoliaceae, and many other plant families. It is difficult to produce alkaloids on a large scale under the strict control of secondary metabolism in plants, and they are too complex for cost-effective chemical synthesis. By using a system that combines microbial and plant enzymes to produce desired benzylisoquinoline alkaloids, we synthesized (S)-reticuline, the key intermediate in benzylisoquinoline alkaloid biosynthesis, from dopamine by crude enzymes from transgenic Escherichia coli. The final yield of (S)-reticuline was 55 mg/liter within 1 h. Furthermore, we synthesized an aporphine alkaloid, magnoflorine, or a protoberberine alkaloid, scoulerine, from dopamine via reticuline by using different combination cultures of transgenic E. coli and Saccharomyces cerevisiae cells. The final yields of magnoflorine and scoulerine were 7.2 and 8.3 mg/liter culture medium. These results indicate that microbial systems that incorporate plant genes cannot only enable the mass production of scarce benzylisoquinoline alkaloids but may also open up pathways for the production of novel benzylisoquinoline alkaloids.
authors.affiliation: Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi-machi, Ishikawa 921-8836, Japan.
authors.forename: Hiromichi
authors.initials: H
authors.lastname: Minami
authors.forename: Ju-Sung
authors.initials: JS
authors.lastname: Kim
authors.forename: Nobuhiro
authors.initials: N
authors.lastname: Ikezawa
authors.forename: Tomoya
authors.initials: T
authors.lastname: Takemura
authors.forename: Takane
authors.initials: T
authors.lastname: Katayama
authors.forename: Hidehiko
authors.initials: H
authors.lastname: Kumagai
authors.forename: Fumihiko
authors.initials: F
authors.lastname: Sato
mesh_terms: D001060:Aporphines / Q000378:metabolism; D044182:Benzylisoquinolines / Q000378:metabolism*; D001600:Berberine Alkaloids / Q000378:metabolism; D001709:Biotechnology / Q000379:methods*; D039323:Coptis / Q000201:enzymology / Q000235:genetics*; D004298:Dopamine / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics*; D017343:Genes, Plant; D016982:Micrococcus luteus / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D019076:Transgenes
chemical_list: D001060:Aporphines; D044182:Benzylisoquinolines; D001600:Berberine Alkaloids; C010941:discretamine; C001670:magnoflorine; D004298:Dopamine; C003298:reticuline