The Extraction and Purification of Bacterial Agglutinating Factor from Carrot (Daucus carota)

Abstract

A phosphate buffered saline extract of carrot (Daucus carota) was found to contain a substance which agglutinated oral bacteria. Several methods for purifying the substance were investigated: ammonium sulfate precipitation, ethanol precipitation, cetylpyridinium chloride precipitation and gel chromatography. The purification methods were monitored by cell agglutination, hemagglutination, chemical analysis and 1-igh pressure liquid chromatography. Chopped carrot pieces were homogenized in a blender and the homogenates filtered through Miracloth and centrifuged. Ammonium sulfate was used to precipitate extraneous proteins. The 100% saturated supernatant contained an active bacterial agglutinating factor. This supernatant was treated with ethanol to precipitate a high molecular weight agglutinating substance that was chiefly carbohydrate and which was stable over wide pH and temperature ranges.