Metabolic engineering to produce phytochromes with phytochromobilin, phycocyanobilin, or phycoerythrobilin chromophore in Escherichia coli.
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.
Mukougawa
Keiko
K
Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan.
Kanamoto
Hirosuke
H
Kobayashi
Toshikazu
T
Yokota
Akiho
A
Kohchi
Takayuki
T
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*
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