Premium
Expression of recombinant full‐length plant phytochromes assembled with phytochromobilin in Pichia pastoris
Author(s) -
Shin Ah-Young,
Han Yun-Jeong,
Song Pill-Soon,
Kim Jeong-Il
Publication year - 2014
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2014.05.050
Subject(s) - pichia pastoris , recombinant dna , phytochrome , biology , botany , pichia , chemistry , biochemistry , microbiology and biotechnology , gene , red light
We have successfully developed a system to produce full‐length plant phytochrome assembled with phytochromobilin in Pichia pastoris by co‐expressing apophytochromes and chromophore biosynthetic genes, heme oxygenase ( HY1 ) and phytochromobilin synthase ( HY2 ) from Arabidopsis . Affinity‐purified phytochrome proteins from Pichia cells displayed zinc fluorescence indicating chromophore attachment. Spectroscopic analyses showed absorbance maximum peaks identical to in vitro reconstituted phytochromobilin‐assembled phytochromes, suggesting that the co‐expression system is effective to generate holo‐phytochromes. Moreover, mitochondria localization of the phytochromobilin biosynthetic genes increased the efficiency of holophytochrome biosynthesis. Therefore, this system provides an excellent source of holophytochromes, including oat phytochrome A and Arabidopsis phytochrome B.