Fig. S1. The iron content in the cytosol of cell in mutant (Mu21-415), complemented strain (Mu21C) and wild type (WT-415). All M. gryphiswaldense strains (wild-type, mutants and complemented strains) were cultured in SLM I for 24 h until stationary phase. When the optical density at 600 nm reached 0.6, 0.3 mM IPTG was added to the culture of complemented strains. 1 l culture was centrifuged respectively at 7000 g for 5 min. The cells were washed by Tris-HCl buffer (20 mM, 4 mM EDTA, pH 7.4) three times and then resuspended by Tris-HCl buffer without EDTA (20 mM, pH 7.4). Suspension of cells (0.34 g ml−1) was disrupted by ultrasonic (200 w, 100 times) and centrifuged at 12 000 g for 20 min. The supernatant was used for ferric ion estimation. Ferric ion concentration was determined as described previously [Tamura H et al. Talanta 1974, 21(4):314–318] with modification as follows. To 100 μl of sample, 250 μl of 10% hydroxylamine hydrochloride and 20 μl 0.25% 1, 10-phenanthroline were added. After 15 min, the absorbance of sample solutions was determined spectrophotometrically at 562 nm.

Table S1. Bacterial strains used in this study.

EMI_2707_sm_FigS1-TabS1.doc36KSupporting info item

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