Material produced by growing Pasteurella tularensis and required for the initiation of growth of P. tularensis from small inocula (growth-initiating substance, GIS) was purified by Sephadex gel filtration and ion-exchange resin chromatography. The purified material was characterized as an anionic compound of low molecular weight, heat-stable at neutral pH and in dilute alkali but destroyed by heating in dilute acid. Out of large variety of known nutrients tested, only iron salts and some iron-chelating compounds (sider-amines) replaced partially GIS in supporting growth of different P. tularensis strains from small inocula. GIS formed complexes with iron and copper ions. The production of GIS by P. tularensis in a chemically defined medium was enhanced by added ornithine. Isotopic evidence established the role of ornithine as a biosynthetic precursor of GIS.
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