title: Metabolic engineering to produce phytochromes with phytochromobilin, phycocyanobilin, or phycoerythrobilin chromophore in Escherichia coli.
abstract: By co-expression of heme oxygenase and various bilin reductase(s) in a single operon in conjunction with apophytochrome using two compatible plasmids, we developed a system to produce phytochromes with various chromophores in Escherichia coli. Through the selection of different bilin reductases, apophytochromes were assembled with phytochromobilin, phycocyanobilin, and phycoerythrobilin. The blue-shifted difference spectra of truncated phytochromes were observed with a phycocyanobilin chromophore compared to a phytochromobilin chromophore. When the phycoerythrobilin biosynthetic enzymes were co-expressed, E. coli cells accumulated orange-fluorescent phytochrome. The metabolic engineering of bacteria for the production of various bilins for assembly into phytochromes will facilitate the molecular analysis of photoreceptors.
authors.affiliation: Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Mukougawa
authors.forename: Hirosuke
authors.initials: H
authors.lastname: Kanamoto
authors.forename: Toshikazu
authors.initials: T
authors.lastname: Kobayashi
authors.forename: Akiho
authors.initials: A
authors.lastname: Yokota
authors.forename: Takayuki
authors.initials: T
authors.lastname: Kohchi
mesh_terms: D001419:Bacteria / Q000378:metabolism; D001654:Bile Pigments / Q000096:biosynthesis; D001664:Biliverdine / Q000031:analogs & derivatives; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000096:biosynthesis*; D005456:Fluorescent Dyes / Q000138:chemical synthesis; D006419:Heme Oxygenase (Decyclizing) / Q000378:metabolism; D052980:Phycobilins; D010798:Phycocyanin; D010799:Phycoerythrin; D010834:Phytochrome / Q000096:biosynthesis*; D015202:Protein Engineering / Q000379:methods*; D045725:Tetrapyrroles / Q000096:biosynthesis*
chemical_list: D001654:Bile Pigments; D029968:Escherichia coli Proteins; D005456:Fluorescent Dyes; D052980:Phycobilins; D045725:Tetrapyrroles; D010798:Phycocyanin; D010799:Phycoerythrin; D010834:Phytochrome; C075811:phytochromobilin; C011070:phycoerythrobilin; C011069:phycocyanobilin; D006419:Heme Oxygenase (Decyclizing); D001664:Biliverdine