Xanthoferrin Siderophore Estimation from the Cell-free Culture Supernatants of Different Xanthomonas Strains by HPLC

Sheo Shankar Pandey; Prashantee Singh; Biswajit Samal; Raj Kumar Verma; Subhadeep Chatterjee

doi:10.21769/BioProtoc.2410

Xanthoferrin Siderophore Estimation from the Cell-free Culture Supernatants of Different Xanthomonas Strains by HPLC

Bio Protoc. 2017 Jul 20;7(14):e2410. doi: 10.21769/BioProtoc.2410.

Authors

Sheo Shankar Pandey^{1

2}, Prashantee Singh^{1

2}, Biswajit Samal^{1

2}, Raj Kumar Verma^{1

2}, Subhadeep Chatterjee¹

Affiliations

¹ Centre for DNA Fingerprinting and Diagnostics, Nampally, Hyderabad-500001, India.
² Graduate studies, Manipal University, Manipal, India.

Abstract

Xanthomonads can scavenge iron from the extracellular environment by secreting the siderophores, which are synthesized by the proteins encoded by xss (Xanthomonas siderophore synthesis) gene cluster. The siderophore production varies among xanthomonads in response to a limited supply of iron where Xanthomonas campestris pv. campestris (Xcc) produces less siderophores than Xanthomonas oryzae pv. oryzae (Xoo) and Xanthomonas oryzae pv. oryzicola (Xoc). Siderophore production can be measured by HPLC and with the CAS (Chrome azurol S)-agar plate assay, however HPLC is a more accurate method over CAS-agar plate assay for siderophore quantification in Xanthomonads. Here we describe how to quantify siderophores from xanthomonads using HPLC.

Keywords: Amberlite XAD-16 resin columns; Bacterial siderophore estimation; HPLC; Xanthoferrin; Xanthomonas siderophores.