Cell-Free Synthesis of Functional Proteins Using Transcription/Translation Machinery Entrapped in Silica Sol-Gel Matrix

Abstract

Herewith we report the encapsulation of functional protein synthesis machinery in a silica sol-gel matrix. When the sol-gel reaction using alkoxysilane monomers was carried out in the presence of Escherichia coli cell extract, macromolecular protein synthesis machinery it the cell extract was successfully immobilized within a silica gel matrix, catalyzing the translation of co-immobilized DNA when supplied with small-molecular-weight substrates for protein synthesis. The efficiency of protein synthesis was affected by the pore size of the gel Structure, which was controlled through the use of appropriate additives during the sol-get reactions, To the best of our knowledge, this is the first report describing the reproduction of the entire set of cornplicated biological process within an inorganic gel matrix, and we expect that the developed technology will find many applications in numerous fields such as high-throughput gene expression and the development of multifunctional biosensors. Biotechnol. Bioeng. 2009;102: 303-307. (C) 2008 Wiley Periodicals, Inc.