The carboxylate type siderophore rhizoferrin and its analogs produced by directed fermentation

Summary Rhizoferrin is a novel carboxylate-type siderophore which has recently been isolated fromRhizopus microsporus and other fungi of the Mucorales (Zygomycetes). The present investigation shows that a variety of rhizoferrin analogs can be produced by directed fermentation. Thus both the diaminobutane backbone and the citric acid side chains of rhizoferrin have been substituted by diamine and citric acid analogs added to the culture medium. The new ligands as well as their iron complexes have been characterized by physicochemical methods. Conditions of precursor incorporation and implications for the biosynthesis of the new siderophores are discussed.     

Rhizoferrin: A complexone type siderophore of the mocorales and entomophthorales (Zygomycetes)

The present investigation presents evidence that rhizoferrin, a novel polycarboxylate or complexone-type siderophore, originally isolated from Rhizopus microsporus, represents the common siderophore within the Zygomycetes. Thus, rhizoferrin could be detected by HPLC analysis in various families of the Mucorales, e.g., Rhizopus microsporus var. rhizopodiformis, Mucor mucedo and Phycomyces nitens (Mucoraceae), Chaetostylum fresenii and Cokeromyces recurvatus (Thamnidiaceae), Cunninghamella elegans and Mycotypha africana (Choanephoraceae) and Mortierella vinacea (Mortierellaceae) and in Basidiobolus microsporus (Entomophthorales). The function of rhizoferrin as a siderophore in the fungus R. microsporus var. rhizopodiformis was demonstrated by time- and concentration-dependent uptake of [55Fe]-labelled rhizoferrin, yielding saturation kinetics with values of Km = 8 microM and V(max) = 1.2 nmol min-1 (mg dry wt)-1.     

Rhizoferrin — a novel siderophore from the fungusRhizopus microsporus var.rhizopodiformis

Summary From a strain ofRhizopus microsporus var.rhizopodiformis a novel siderophore, named rhizoferrin, was isolated by ion-exchange column chromatography, gel filtration and preparative HPLC. Hydrolysis with 6 M HCl and subsequent gas chromatography/mass spectrometry (GUMS) of the esterified/trifluoroacetylated derivatives indicated that citric acid and diaminobutane were the only constituents. From positive fastatom-bombardment (FAB) and ion-spray tandem mass spectrometry, a molecular mass of 436 Da and the assignment of several daughter ion fragments could be obtained, which indicated the presence of two citric acid residues and one diaminobutane residue. NMR studies finally confirmedN ¹,N ⁴-bis(1-oxo-3-hydroxy-3,4-dicarboxybutyl)-diaminobutane as the structure of rhizoferrin. The iron-binding property was demonstrated on chromeazurol S plates and its siderophore activity was confirmed by iron transport measurements in young mycelia ofR. microsporus. While rhizoferrin and also ferrioxamines B and E proved to be effective siderophores, coprogen was a poor siderophore in this fungus.