Use of Highly Porous Bead Cellulose with Attached Bacitracin for Affinity Chromatography of a Microbial Proteinase

Abstract

Although affinity chromatography is currently used as a preparative method on the laboratory scale, it is expected that its principle will be utilized still more for large scale preparations of many proteins of practical importance. For this purpose the requirements imposed on the adsorbent, the method of its immobilization, and the carrier are very high. A decisive role is played by the ligand. The cyclic nonapeptide bacitracin (Figure 1), a noncompetitive inhibitor of many proteolytic enzymes, was proved to be an efficient ligand for their purification by affinity chromatography. It was attached to agarose or silichrom by cyanogen bromide and benzoquinon, respectively (Stepanov et al., 1981). Both carriers and their activation have some shortcomings which can be avoided by using bead cellulose as a carrier and a different method of immobilization of the ligand.