Purification of Pyoverdines of Pseudomonas fluorescens 2-79 by Copper-Chelate Chromatography

• DOI: 10.1128/aem.61.11.3769-3774.1995
• Publication Date: 2020-01-06T17:58:34Z
• Authors: R Xiao; W S Kisaalita
• Keywords: Pyoverdine; Absorbance; Pseudomonas fluorescens

Three pyoverdines, Pf-A, Pf-B, and Pf-C, were purified with copper-chelate Sepharose Sephadex G-15 columns from Pseudomonas fluorescens 2-79, the yields (per 100 ml of culture supernatant) 2.8, 21.6, 3.2 mg, respectively. The absorption fluorescence spectra these pyoverdines strongly pH dependent. Characteristic changes in maximal absorbance wavelengths observed when Fe(sup3+) or Cu(sup2+) was added. addition shifted pyoverdine Pf-B spectrum so that it exhibited a single peak at 410 nm but did not give rise to new maximum approximately 460 nm, which appeared Fluorescence quenching experiments revealed forward reaction rate constant much higher (10(sup4) 10(sup5) M(sup-1) s(sup-1)) than (10(sup2) s(sup-1)). However, Cu(sup2+)-pyoverdine complexes completely dissociated by EDTA low concentration (0.1 mM), while level Fe(sup3+)-pyoverdine complex dissociation same relatively low. Formation free three types incubated separately P. 2-79 cells, thus demonstrating Pf-C mediate iron transport.