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The flow-through fractions were pooled and concentrated by centrifugation with a 10 kDa cutoff and subjected to size exclusion chromatography on a Superdex-200 column in Buffer C (25 mM Tris-HCl, pH 8.0, 500 mM NaCl, 2 mM EDTA) and monomeric fractions pooled

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it is part of the prosthetic group of enzymes involved in respiration and DNA synthesis and is part of the active center of proteins devoted to oxygen and electron transport, such as hemoglobin (Hb) and cytochromes (Cy), respectively