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Cell-free translation in reversed micelles
S N Nametkin1, M I Kolosov, Ovodov SYu
1Department of Chemical Enzymology, Faculty of Chemistry, M. V. Lomonosov Moscow State University, Russia.
FEBS Letters
|September 14, 1992
Summary
Cell-free protein synthesis was achieved in reversed micelles (RM) using a wheat germ system. This method shows promise for producing hydrophobic proteins, with yields comparable to aqueous solutions.
Area of Science:
- Biochemistry
- Molecular Biology
- Biotechnology
Background:
- Cell-free protein synthesis offers advantages for producing specific proteins.
- Reversed micelles (RM) in organic solvents can encapsulate biological systems.
- Hydrophobic protein synthesis presents unique challenges in vitro.
Purpose of the Study:
- To demonstrate cell-free translation within reversed micelles (RM).
- To evaluate the efficiency of synthesizing human interleukin-2 using a wheat germ system in RM.
- To assess the potential of RM as a medium for hydrophobic protein synthesis.
Main Methods:
- Utilized a wheat germ cell-free translation system solubilized in Brij 96 reversed micelles (RM) in cyclohexane.
- Synthesized human interleukin-2 as a model protein.
- Recovered translation components and product via acetone precipitation.
- Investigated the effect of surfactant hydration on recovery and yield.
Main Results:
- Achieved cell-free translation in Brij 96 RM with yields comparable to aqueous solutions.
- Demonstrated successful recovery of system components and product.
- Found that translation yields are dependent on the degree of surfactant hydration.
- Observed no translation in Aerosol OT RM, likely due to Mg2+ ion binding.
Conclusions:
- Reversed micelles (RM) provide a viable medium for cell-free protein synthesis.
- The Brij 96 RM system is a promising platform for the cell-free synthesis of hydrophobic proteins.
- Surfactant properties, such as Mg2+ ion binding, significantly impact translation efficiency in RM.