title: Biosynthesis of soluble carotenoid holoproteins in Escherichia coli.
abstract: Carotenoids are widely distributed natural pigments that are excellent antioxidants acting in photoprotection. They are typically solubilized in membranes or attached to proteins. In cyanobacteria, the photoactive soluble Orange Carotenoid Protein (OCP) is involved in photoprotective mechanisms as a highly active singlet oxygen and excitation energy quencher. Here we describe a method for producing large amounts of holo-OCP in E.coli. The six different genes involved in the synthesis of holo-OCP were introduced into E. coli using three different plasmids. The choice of promoters and the order of gene induction were important: the induction of genes involved in carotenoid synthesis must precede the induction of the ocp gene in order to obtain holo-OCPs. Active holo-OCPs with primary structures derived from several cyanobacterial strains and containing different carotenoids were isolated. This approach for rapid heterologous synthesis of large quantities of carotenoproteins is a fundamental advance in the production of antioxidants of great interest to the pharmaceutical and cosmetic industries.
authors.affiliation: 1] Commissariat à l'Energie Atomique (CEA), Institut de Biologie et Technologies de Saclay (iBiTec-S), 91191 Gif sur Yvette, France [2] Centre National de la Recherche Scientifique (CNRS), UMR 8221, 91191 Gif sur Yvette, France [3] Phycosource, 13 boulevard de l'Hautil, 95092 Cergy Cedex, France.
authors.forename: Céline
authors.initials: C
authors.lastname: Bourcier de Carbon
authors.affiliation: 1] Commissariat à l'Energie Atomique (CEA), Institut de Biologie et Technologies de Saclay (iBiTec-S), 91191 Gif sur Yvette, France [2] Centre National de la Recherche Scientifique (CNRS), UMR 8221, 91191 Gif sur Yvette, France.
authors.forename: Adrien
authors.initials: A
authors.lastname: Thurotte
authors.affiliation: 1] Commissariat à l'Energie Atomique (CEA), Institut de Biologie et Technologies de Saclay (iBiTec-S), 91191 Gif sur Yvette, France [2] Centre National de la Recherche Scientifique (CNRS), UMR 8221, 91191 Gif sur Yvette, France.
authors.forename: Adjélé
authors.initials: A
authors.lastname: Wilson
authors.affiliation: 1] INRA, Institut Jean-Pierre Bourgin, UMR 1318, ERL CNRS 3559, Saclay Plant Sciences, RD10, F-78026 Versailles, France [2] AgroParisTech, Institut Jean-Pierre Bourgin, UMR 1318, ERL CNRS 3559, Saclay Plant Sciences, RD10, F-78026 Versailles, France.
authors.forename: François
authors.initials: F
authors.lastname: Perreau
authors.affiliation: 1] Commissariat à l'Energie Atomique (CEA), Institut de Biologie et Technologies de Saclay (iBiTec-S), 91191 Gif sur Yvette, France [2] Centre National de la Recherche Scientifique (CNRS), UMR 8221, 91191 Gif sur Yvette, France.
authors.forename: Diana
authors.initials: D
authors.lastname: Kirilovsky
mesh_terms: D000975:Antioxidants / Q000378:metabolism; D002338:Carotenoids / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D055668:Photochemical Processes; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D000975:Antioxidants; D029968:Escherichia coli Proteins; D011993:Recombinant Fusion Proteins; D002338:Carotenoids