A new procedure for the removal of iron from complex media is given. With this method, the iron requirements for the growth of in casein hydrolysate medium were investigated. Trypticase medium (0.6-0.8 μg. Fe/ml.) was depleted of iron by treatment with the specific iron-chelator bathophenanthroline; on analysis it was shown to contain 0.01-0.02 μg. Fe/ml. Preliminary studies showed that grew well in the iron-depleted medium. Further iron restriction of the already depleted medium was accomplished by the addition of conalbumin, a specific iron-chelating protein present in egg white. The initiation of growth, rate of growth, and total crop were all dependent on the concentration of free iron in the medium. The availability of iron was a function of the percentage iron saturation of the conalbumin.


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