Nitrogen-Containing and Carbohydrate-Containing Antigen from Actinomyces bovis

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Abstract
Water-soluble, heat-stable antigens have been isolated from the supernatant fluids of broth cultures of Actinomyces bovis ATCC 10048 after 8 days of growth in a broth medium composed of Casamino Acids, yeast extract, Tween 80 (polyoxyethylene sorbitan monooleate), sodium thio-glycolate, dextrose, and sodium chloride. The antigens were precipitated from the culture supernatant fluid with alcohol and purified by fractional precipitation with alcohol in the presence of calcium or zinc and by chromatography on diethylaminoethyl Sephadex. Two serologically active substances which differed in chemical composition and size were characterized. The larger one contained 71% hexose and 2.8% nitrogen, and the smaller one contained 45% hexose and 5.4% nitrogen. Heterogeneity of these fractions could not be demonstrated by electro-phoresis in free solution at pH 7.8 or 2, by ultracentrifugal analysis, by double diffusion in agar, or by immunoelectrophoresis. Despite their differences in chemical composition and size, they appeared identical in activity as antigens in complement fixation and in double-diffusion tests in agar. Each was found to contain mannose as its chief component after hydrolysis and paper chromatography with three solvent systems. A small percentage of the total nitrogen may be attributed to the presence of amino sugars but the remainder is as yet unidentified.