Purification of Histamine Sensitizing Factor of Bordetella Pertussis

Abstract

rurification of histamine sensitizing factor (HSP) of Bordetella pertussis was attempted from two sources : i.e. sonicate of the bacterial cells and bho culture supernatant. 1. Subcollular fractions were prepared from the cells disintegrated by sonic oscillation. A fraction sedimentable at 195,000 x g for 90 minutes contained potent HSF activity as well as lethal toxicity. This fraction was mainly composed of ribosome particle. 2. Electrophoretic experiments and ultracentrifigal analysis of the ribosomal fraction digested with protease or ribonuclease indicated that HSF activity can not be refored to the ribosome particle itself. 3. HSF could be extracted from the ribosomal fraction with surface activa agents such as sodium deoxycholate. Lethal toxicity was completely disappeared by this treatment. 4. Several methods for purification of HSF from the culture supernatant, such as density gradient electrophoresis,.PH gradient clution and polyvinyl pyrrolidone treatment, were surveyed.