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Figure 1
Analysis of recombinant NiR. (a) SDS-PAGE containing 12.5% (w/v) acrylamide of whole cell extracts of BL21(DE3) containing pEnirsp1 grown at 303 K in a 1 l fermenter. Induction at OD600nm ≃ 0.5: lane S, molecular mass markers; lane 1, negative control, BL21(DE3); lane 2, sample taken immediately after addition of 0.25 mM IPTG; lanes 3 to 12, samples taken at 30 min intervals after induction. (b) EPR spectra of recombinant NiR in 20 mM MES buffer, pH 6.0. Spectrum 1. As-purified protein (T2D form) at 9.4 mg ml−1 (255 µM monomer). Spectrum 2. Protein incubated with 1 mM CuSO4 after purification (partially reconstituted form) at 6.1 mg ml−1 (165 µM monomer). Spectrum 3. Protein purified after incubation of the periplasmic fraction with CuSO4 (fully reconstituted form) at 4.8 mg ml−1 (135 µM monomer). The spectra were recorded at 60 K at a microwave frequency of 9.312 GHz and normalized for ease of comparison. The type-1 and type-2 features are indicated.

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ISSN: 1600-5775
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