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Affinity Purification of a Siderophore That Exhibits an Antagonistic Effect against Soft Rot Bacterium

Mohamed Helmy*, Doa Baddar, and Mohamed Hisham El'Masry

Department of Biotechnology, Institute of Graduate Studies and Research (IGSR), Alexandria University, 163 El Horreya Avenue, El Chatby 21526, Alexandria, Egypt; fax: 00203-4285792; E-mail: esmailgadmoh@yahoo.fr

* To whom correspondence should be addressed.

Received August 8, 2007; Revision received January 5, 2008
Bacterial colonies were isolated from different Egyptian soil samples. From these isolates, one bacterial species was found to produce siderophore. Using classical and biochemical identification methods, the siderophore producing isolate was identified as Pseudomonas fluorescens. Based on the affinity of siderophores for metal ions, an affinity chromatography system was designed for the purification of the siderophore in one step. It was possible to isolate 25 mg siderophore per liter of culture media. The purified siderophore was found to exist in two forms of approximately 30 and 90 kD. They are believed to be polymers of several siderophore molecules. Both forms were found to be active against the pathogen Erwinia carotovora var. carotovora, the causal bacteria of soft rot disease on potato tubers. The advantage of this method over other purification methods is that it uses metal ion so it can be applied for the purification of the known types of siderophores. Moreover, the purification is based on affinity chromatography, so the siderophore purity state permits several biotechnological applications without further treatments.
KEY WORDS: affinity chromatography, Erwinia carotovora, siderophore, soft rot bacterium

DOI: 10.1134/S0006297908070055