Abstract
Rabbit antiserum to the purified preparation of adenylate cyclase of Brevibacterium liquefaciens was obtained. A solid phase immunoadsorbent prepared by the cyanogen bromide procedure was shown to bind adenylate cyclase of B. liquefaciens. The elution of bound adenylate cyclase was successfully accomplished with 1 mM to 10 mM NaOH containing 50 mM KCl. Starting from a partially purified preparation, adenylate cyclase was purified 125-fold in the single step of affinity chromatography with a yield of 54%. However, when the partially purified adenylate cyclase was loaded on the immunoadsorbent over the adsorbing capacity, the bound adenylate cyclase was recovered almost quantitatively and approximately 400-fold purification was achieved by this single step procedure. The overall purity was estimated to be 25 to 30% of the homogeneous preparation.
The immunoadsorbent prepared with antiserum to adenylate cyclase of B. liquefaciens was also applicable to the partial purification of adenylate cyclase from Micrococcus lysodeikticus
