Figure 4.

Final Sephadex G-15 column purification of the partially purified ninhydrin-reactive compound recovered from preparative TLC chromatograms. A sterile aqueous solution containing the partially purified SBW25 ninhydrin-reactive compound was prepared by extraction of the appropriate zone of preparative TLC chromatograms as described in the Methods section. This solution was taken to dryness in vacuo, and the recovered solids were dissolved in 5 mL of deionized water for application to the Sephadex G-15 column. The column was eluted with deionized water. Fractions (5 mL each) were collected and analyzed for reaction with the Fe- and Cu-ChromeAzurol S reagents. The fractions corresponding to the Cu-binding peak were pooled (as indicated by the double arrow) and concentrated for structural identification.