THE EXPRESS-METHOD OF OBTAINING PROTEASE ECP32, THE UNIQUE INSTRUMENT IN ACTIN INVESTIGATIONS
A. V. Morozova,1 S. Yu. Khaitlina, A. Yu. Malinin
Institute of Cytology RAS, St. Petersburg;
1 e-mail: avmoro@gmail.com
The protease ECP32 is significant in investigations of actin, the basic protein of muscles and the cytoskeleton. The enzyme originates from the natural
enterobacteria strain, which accumulates minor amounts of the protease intracellularly at the post-exponential growth phase. The limiting factor for biosynthesis is
the amount of oxygen that has entered the medium. The highly effective method of two-phase cultivation with vigorous aeration at the exponential growth phase was
recommended. Based upon the enzyme properties studied, there is a decreased potential for success when using either the affinity or one-stage purification methods.
In order to overcome obstacles in the aforementioned methods, a simple method for ECP32 preparation and storage was developed, with purity and activity levels
satisfying the requirements of actin structure and function investigations.
Key words: actin, ECP32 protease, limited proteolysis, post-logaryhmic synthesis, bacterial intracellular enzymes, protein purification
Back
Contents
Main
