title: Synthesis of phycocyanobilin in mammalian cells.
abstract: The chromophore 3-Z phycocyanobilin (PCB, (2R,3Z)-8,12-bis(2-carboxyethyl)-18-ethyl-3-ethylidene-2,7,13,17-tetramethyl-2,3-dihydrobilin-1,19(21H,24H)-dione) mediates red and far-red light perception in natural and synthetic biological systems. Here we describe a PCB synthesis strategy in mammalian cells. We optimize the production by co-localizing the biocatalysts to the substrate source, by coordinating the availability of the biocatalysts and by reducing the degradation of the reaction product. We show that the resulting PCB levels of 2 μM are sufficient to sustain the functionality of red light-responsive optogenetic tools suitable for the light-inducible control of gene expression in mammalian cells.
authors: Faculty of Biology, University of Freiburg, Schänzlestrasse 1, 79104 Freiburg, Germany. wilfried.weber@biologie.uni-freiburg.de.
authors: Konrad
authors: K
authors: Müller
authors: Raphael
authors: R
authors: Engesser
authors: Jens
authors: J
authors: Timmer
authors: Ferenc
authors: F
authors: Nagy
authors: Matias D
authors: MD
authors: Zurbriggen
authors: Wilfried
authors: W
authors: Weber
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D055162:Biocatalysis; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D000458:Cyanobacteria / Q000201:enzymology; D051547:Heme Oxygenase-1 / Q000235:genetics / Q000378:metabolism; D008027:Light; D044925:Oxidoreductases Acting on CH-CH Group Donors / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D052980:Phycobilins / Q000096:biosynthesis* / Q000737:chemistry; D010798:Phycocyanin / Q000096:biosynthesis* / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D052980:Phycobilins; D034741:RNA, Small Interfering; D010798:Phycocyanin; C011069:phycocyanobilin; D051547:Heme Oxygenase-1; D044925:Oxidoreductases Acting on CH-CH Group Donors; C019789:biliverdin reductase