Purification of Biotlnylated Enzymes Using Streptavidin Mutean Matrix

摘要

Wild-type streptavidin is known to interact with biotin more or less irreversibly [1]. This hampers use of streptavidin as a ligand for affinity purification matrices. Therefore, recombinant streptavidin technology was used to screen for a mutant streptavidin with reduced biotin affinity. By substitution of three amino acids, a mutein with a biotin dissociation constant of 1.3 * 10~(-7) M was obtained [2]. The immobilized mutein, the Streptavidin Mutein Matrix, proved to be a convenient and stable affinity matrix, enabling purification of biotinylaLed pro-teins with excellent purity and good recovery.