title: A new hybrid protein for production of recombinant bacteriorhodopsin in Escherichia coli.
abstract: Unique properties of bacteriorhodopsin, namely, photochromism and high thermal stability, make this protein an attractive target for physico-chemical studies, as well as for various biotechnological applications. Using Mistic as a suitable carrier for insertion of recombinant membrane proteins into cytoplasmic membrane of Escherichia coli, we developed a system for overexpression of bacteriorhodopsin and worked out an efficient procedure for its purification and renaturation with the final yield of 120 mg/l of refolded protein, which is the highest value reported to date for bacteriorhodopsin produced in E. coli. Functional activity of recombinant bacteriorhodopsin was confirmed by spectroscopic and electrochemical assays.
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow 117997, Russia. genchem@ibch.ru
authors.forename: Oksana V
authors.initials: OV
authors.lastname: Nekrasova
authors.forename: Andrey N
authors.initials: AN
authors.lastname: Wulfson
authors.forename: Roman V
authors.initials: RV
authors.lastname: Tikhonov
authors.forename: Sergey A
authors.initials: SA
authors.lastname: Yakimov
authors.forename: Tatyana N
authors.initials: TN
authors.lastname: Simonova
authors.forename: Anna I
authors.initials: AI
authors.lastname: Tagvey
authors.forename: Dmitry A
authors.initials: DA
authors.lastname: Dolgikh
authors.forename: Mikhail A
authors.initials: MA
authors.lastname: Ostrovsky
authors.forename: Mikhail P
authors.initials: MP
authors.lastname: Kirpichnikov
mesh_terms: D000222:Adaptation, Physiological / Q000528:radiation effects; D001436:Bacteriorhodopsins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D001681:Biological Assay; D001709:Biotechnology / Q000379:methods*; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000378:metabolism*; D019617:Halobacterium salinarum / Q000378:metabolism*; D008027:Light; D020673:Protein Renaturation / Q000528:radiation effects; D017433:Protein Structure, Secondary; D017494:Proton Pumps / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D017494:Proton Pumps; D011994:Recombinant Proteins; D001436:Bacteriorhodopsins