Cell‐free translation in reversed micelles

FEBS Letters

Alexandrov, Alexander N.³  Ovodov, Sergey Yu.²  Kolosov, Michael I.³  Kabanov, Alexander V.¹  Levashov, Andrey V.¹  Nametkin, Sergey N.¹  Alakhov, Valery Yu.³ 

[1] Department of Chemical Enzymology, Faculty of Chemistry, M.V. Lomonosov Moscow State University, Moscow 119899, Russia;Institute of Protein Research, Russian Academy of Sciences, Pushchino 142292, Moscow Region, Russia;Research Center of Molecular Diagnostics and Therapy, Simpheropolsky blvd. 8, Moscow 113149, Russia

关键词: Cell-free translation;    Interleukin;    Micellar enzymology;    Reversed micelle;    Aerosol OT;    (sodium bis-(2-ethylhexyl) sulfosuccinate);    Brij 96;    oleyl-poly(10)oxyethylene ether;    DDT;    dithiotreitol;    IL-2;    interleukin 2;    HEPES;    N-(2-hydroxyethyl) piperazine-N′-(2-ethanesulfonic acid);    PAA;    poly(acrylamide);    RM;    reversed micelle;    TCA;    trichloracetic acid;    WGE;    wheat germ extract;   

DOI  :  10.1016/0014-5793(92)80800-V

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Cell-free translation in reversed micelles (RM) of surfactants in organic solvents is demonstrated using as an example the synthesis of human interleukin-2 by the wheat germ translation system solubilized in Brij 96 (oleyl-poly(10)oxyethylene ether) RM in cyclohexane. The translation system components and the product were recovered from the RM system by acetone precipitation. The recovery and translation reaction yields depended on the degree of surfactant hydration. The translation yields in Brij 96 RM were close to that observed in regular aqueous solution. The Brij 96 RM system is regarded as a promising media for the cell-free synthesis of hydrophobic proteins. Meanwhile, no translation reaction was observed in Aerosol OT (sodium bis(2-ethylhexyl) sulfosuccinate) RM in octane, which presumably is due to the ability of Aerosol OT to bind Mg²⁻ ions necessary for the functioning of the translation apparatus.

【 授权许可】

Unknown