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4 protocols using feso4 7h2o

1

Nanocomposite Synthesis for Pollutant Removal

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In the synthesis of nanocomposites, MWCNT (D × L 110–170 nm × 5–9 μm), NiCl2 × 6H2O, ascorbic acid, cetyltrimethylammonium bromide (CTAB) were purchased from Sigma-Aldrich (Schnelldorf, Germany), FeCl3 × 6H2O from Alfa Aesar (Kandel, Germany), FeSO4 × 7H2O from VWR Chemicals (Wien, Austria) and NH3 25% solution from Chemical Company (Iaşi, Romania). Tartrazine was chosen as pollutant in this study and was purchased from Sigma-Aldrich (Schnelldorf, Germany). The pH adjustment was performed with HCl and NaOH that were purchased from Sigma-Aldrich (Schnelldorf, Germany) and VWR Chemicals (Wien, Austria), respectively. Aqueous solutions were prepared using ultrapure water (Direct-Q® 3 UV Water Purification System, Merck, Darmstadt, Germany).
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2

Microalgal Antioxidant Capacity Determination

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The antioxidant capacity of microalgal strains to reduce ferric iron (Fe3+) to ferrous iron (Fe2+) was determined by the FRAP assay, as previously reported [39 (link)]. Briefly, a solution containing 300 mM acetate buffer (pH 3.6), 10 mM 2,4,6-Tri(2-pyridyl)-s-triazine (TPTZ) in 40 mM HCl, and 20 mM FeCl3·6H2O (VWR, Milan, Italy) was added to each microalgal extract (85 μL). Following 30 min incubation at room temperature, the absorbance was measured at 593 nm using a Perkin-Elmer Lambda 365 spectrophotometer (Perkin Elmer Italia, Milano, Italy), and results were expressed as Fe2+ equivalents (μM) using a water solution of FeSO4·7H2O (100–2000 μM; VWR, Milan, Italy) used as a standard. All reagents were purchased from Fluka-Sigma-Aldrich, Inc. (St. Louis, MO, USA), unless otherwise specified.
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Holoferritins Rendered Iron-Free

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Holoferritins (i.e. iron containing ferritin) were rendered iron-free by dialysis against sodium hydrosulfite (dithionite), Na2S2O4, and complexation with 2,2′-bipyridyl at pH 6.0. Protein concentrations were determined spectrophotometrically using a molar absorptivity of 24000 cm−1 M−1 at 280 nm for the 24-mer apoprotein (iron-free protein) (35 (link)). All chemicals were reagent grade and used without further purification. Mops (3-(N-morpholino)-propanesulfonic acid) buffer was purchased from Research Organics (Cleveland, OH) and FeSO4·7H2O from J. T. Baker (Phillipsburg, NJ). Sodium dithionite, Na2S2O4 ferrozine and 2,2′-bipyridyl were purchased from Sigma-Aldrich (St. Louis, MO). Ferrous sulfate stock solutions were freshly prepared immediately before each experiment in a dilute HCl solution at pH 2.0.
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4

Iron-free Holoferritin Preparation

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Holoferritins (iron containing ferritin) were rendered iron-free by dialysis against sodium hydrosulfite (dithionite), Na2S2O4, and complexation with 2,2′-bipyridyl at pH 6.0.24 (link) Protein concentrations were determined using the Advanced Protein Assay (http://cytoskeleton.com) or spectrophotometrically using a molar absorptivity of 24000 cm−1 M−1 at 280 nm for the 24-mer apoprotein (iron-free protein, this work). All chemicals were reagent grade and used without further purification. Mops (3-(N-morpholino)-propanesulfonic acid) buffer was purchased from Research Organics (Cleveland, OH) and FeSO4·7H2O from J. T. Baker (Phillipsburg, NJ). Sodium dithionite, Na2S2O4, and 2,2′-bipyridyl were purchased from Sigma-Aldrich (St. Louis, MO). Fe(II) stock solutions were freshly prepared immediately before each experiment in a dilute HCl solution at pH 2.0. Freshly prepared hydrogen peroxide solutions were assayed either by electrode oximetry using catalase (EC 1.11.1.6, 65000 units/mg, Roche Molecular Biochemicals) by following the amount of O2 produced or by ultraviolet–visible (UV–vis) spectroscopy using a molar absorptivity of 43.6 M−1 cm−1 at 240 nm.24 (link)
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