A new hybrid protein for production of recombinant bacteriorhodopsin in Escherichia coli.
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.
Nekrasova
Oksana V
OV
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow 117997, Russia. genchem@ibch.ru
Wulfson
Andrey N
AN
Tikhonov
Roman V
RV
Yakimov
Sergey A
SA
Simonova
Tatyana N
TN
Tagvey
Anna I
AI
Dolgikh
Dmitry A
DA
Ostrovsky
Mikhail A
MA
Kirpichnikov
Mikhail P
MP
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
D017494:Proton Pumps; D011994:Recombinant Proteins; D001436:Bacteriorhodopsins