Paper chromatography of vitamin B12 and related bacterial growth factors.

In a preliminary note (l), the present authors reported that Lactobacillus Eeichmannii 313 is capable of using six alternative factors for growth in vitamin Bn-deficient media. Two of these were found in vitamin Blz concentrates and presumably represent two forms of the vitamin. Hoffmann et al. (2) previously noted that crystalline antipernicious anemia factor (APA) (vitamin B1.J supports the growth of L. Zeichmannii 313. Of the other four factors, one was provisionally identified as thymidine, a substance which was reported by Snell et al. (3) as supporting the growth of this organism. The three remaining growth factors are of unknown composition. In the present report, a paper chromatographic procedure for separating the alternative growth factors is described as well as the method of recognizing the positions of the several factors on a chromatogram by use of L. Zeichmannii 313 as a microbiological indicator. Extension of the method to the examination of other natural materials has revealed the existence of at least one other substitute growth factor replacing vitamin Bti in the nutrition of the test organism. That the naturally occurring alternative growth factors which replace vitamin Bn in the nutrition of L. Zeichmannii 313 are desoxyribosides would appear probable from evidence presented in this paper. Kitay et al. (4) have shown that desoxyribosides can replace thymidine in the nutrition of several lactic acid bacteria. The paper chromatographic technique has aided in the interpretation of assay values for apparent vitamin Bn activity as measured by the usual tube assay.