Permeabilization of microorganisms by Triton X-100

✍ Scribed by G.F. Miozzari; P. Niederberger; R. Hu¨tter

• Publisher: Elsevier Science
• Year: 1978
• Tongue: English
• Weight: 904 KB
• Volume: 90
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(78)90026-x

✦ Synopsis

A simple permeabilization procedure has been developed which allows the reliable determination of enzyme activities in situ in a variety of different microorganisms. Permeabilization is obtained by freezing cell suspensions in the presence of a low concentration of the anionic detergent Triton X-100. After thawing, the cells can be used directly in the enzyme assays. The procedure has been optimized using the yeast Saccharomyces cerevisiae.

Yeast cells are completely permeabilized by Triton X-100 concentrations of 0.05% (v/v), and permeabilization is independent of cell age and cell concentration. The treatment makes the cells freely diffusible for macromolecules up to molecular weights around 70,000. Cytoplasmic and mitochondrial amino acid biosynthetic enzymes as well as aminoacyl-tRNA synthetases could be readily measured in treated cells. The method has been successfully applied to the determination of enzyme activities in other fungi as well as in gram-positive and gramnegative bacteria.