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Dye affinity chromatography for fast and simple purification of fucoidan from marine brown algae
Authors:Thomas Hahn  Ahmed Zayed  Mariya Kovacheva  Ralf Stadtmüller  Siegmund Lang  Kai Muffler  Roland Ulber
Institution:1. Molecular Biotechnology, Fraunhofer Institute for Interfacial Engineering and Biotechnology, Stuttgart, Germany;2. Institute of Bioprocess Engineering, University of Kaiserslautern, Kaiserslautern, Germany;3. Department of Pharmacognosy, Tanta University, College of Pharmacy, Tanta, Egypt;4. Biotechnology and Bioinformatics, Institute of Biochemistry, University of Braunschweig, Braunschweig, Germany;5. Department of Life Sciences and Engineering, University of Applied Sciences Bingen, Bingen, Germany
Abstract:Fucoidan is a sulfated polysaccharide with promising pharmacological applications. Due to its medicinal properties, there is a demand for a separation technique that yields a high purification grade. Here, we present a novel purification tool for recovering fucoidan from the marine brown macroalgae Fucus vesiculosus. The developed method is based on amino‐derivatized Sepabeads® EC‐EA. The beads were modified with toluidine blue (TB), a thiazine derivative, to exploit the strong donor acceptor interactions between the cationic dye and the anionic polysaccharide. The adsorption kinetics and the binding capacity of the resin were analyzed. A Sips model was used to approximate the adsorption isotherm, resulting in a maximum capacity of 127.7 mg fucoidan per g adsorbent. Investigation of the effect of adsorption step's pH on purity and chemical structure was performed by TB and Fourier transform infra‐red spectroscopy assays. Results showed that adsorption at pH 1 and 6 had negligible effects on fucoidan's chemical structure. However, purity was actually improved by 1.55‐ and 1.69‐fold at pH 1 and 6, respectively, with an average yield of 5 g/100 g dried algae powder. In contrast, only a 1.46‐fold increment was observed in fucoidan purified by the traditional method at pH 2, with a yield of 7.5 g/100 g dried algae powder. Furthermore, fucoidan purified by this method at pH 6 complies with, or even exceeds the quality of the commercially available (≥95% pure) fucoidan (Sigma‐Aldrich®) with respect to molecular weight and sulfur content. Therefore, dye affinity chromatography provides more advantages than the classically used techniques for fucoidan purification.
Keywords:Dye affinity  Fucoidan  One‐step purification  Sepabeads  Toluidine blue O
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