Physicochemical and Biological Characterization of Fucoidan from Fucus vesiculosus Purified by Dye Affinity Chromatography.
A comparative study concerning the physicochemical, monomeric composition and biological characters among different fucoidan fractions is presented. Common purification techniques for fucoidan usually involve many steps. During these steps, the important structural features might be affected and consequently alter its biological activities. Three purified fractions were derived from Fucus vesiculosus water extract which, afterwards, were purified by a recently-developed dye affinity chromatography protocol. This protocol is based on dye-sulfated polysaccharide interactions. The first two fractions were obtained from crude precipitated fucoidan at different pH values of the adsorption phase: pH 1 and 6. This procedure resulted in fucoidan_1 and 6 fractions. The other, third, fraction: fucoidan_M, however, was obtained from a buffered crude extract at pH 1, eliminating the ethanol precipitation step. All of the three fractions were then further evaluated. Results revealed that fucoidan_M showed the highest sulfur content (S%), 12.11%, with the lowest average molecular weight, 48 kDa. Fucose, galactose, and uronic acid/glucose dimers were detected in all fractions, although, xylose was only detected in fucoidan_1 and 6. In a concentration of 10 µg·mL(-1), Fucoidan_6 showed the highest heparin-like anticoagulant activity and could prolong the APTT and TT significantly to 66.03 ± 2.93 and 75.36 ± 1.37 s, respectively. In addition, fucoidan_M demonstrated the highest potency against HSV-1 with an IC50 of 2.41 µg·mL(-1). The technique proved to be a candidate for fucoidan purifaction from its crude extract removing the precipitation step from common purification protocols and produced different fucoidan qualities resulted from the different incubation conditions with the immobilized thiazine toluidine blue O dye.
Institute of Bioprocess Engineering, University of Kaiserslautern, Gottlieb-Daimler-Straße 49, 67663 Kaiserslautern, Germany. zayed@mv.uni-kl.de.|Department of Pharmacognosy, Tanta University, College of Pharmacy, El Guish Street, 8130 Tanta, Egypt. zayed@mv.uni-kl.de.
Ahmed
A
Zayed
Department of Life Sciences and Engineering, University of Applied Sciences Bingen, Berlinstraße 109, 55411 Bingen, Germany. K.Muffler@fh-bingen.de.
Kai
K
Muffler
Department of Molecular Biotechnology, Fraunhofer Institute for Interfacial Engineering and Biotechnology, Nobelstraße 12, 70569 Stuttgart, Germany. thomas.hahn@igb.fraunhofer.de.
Thomas
T
Hahn
Department of Molecular Biotechnology, Fraunhofer Institute for Interfacial Engineering and Biotechnology, Nobelstraße 12, 70569 Stuttgart, Germany. steffen.rupp@igb.fraunhofer.de.
Steffen
S
Rupp
Department of Molecular Biotechnology, Fraunhofer Institute for Interfacial Engineering and Biotechnology, Nobelstraße 12, 70569 Stuttgart, Germany. doris.finkelmeier@igb.fraunhofer.de.
Doris
D
Finkelmeier
Department of Molecular Biotechnology, Fraunhofer Institute for Interfacial Engineering and Biotechnology, Nobelstraße 12, 70569 Stuttgart, Germany. anke.burger-kentischer@igb.fraunhofer.de.
Anke
A
Burger-Kentischer
Institute of Bioprocess Engineering, University of Kaiserslautern, Gottlieb-Daimler-Straße 49, 67663 Kaiserslautern, Germany. ulber@mv.uni-kl.de.
Roland
R
Ulber
D000925:Anticoagulants / Q000737:chemistry / Q000494:pharmacology; D002846:Chromatography, Affinity / Q000379:methods; D044645:Fucus / Q000235:genetics*; D018259:Herpesvirus 1, Human / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D011134:Polysaccharides / Q000737:chemistry* / Q000494:pharmacology*; D013431:Sulfates / Q000737:chemistry
D000925:Anticoagulants; D011134:Polysaccharides; D013431:Sulfates; C007789:fucoidan
anticoagulant; antiviral; dye-affinity chromatography; fucoidan; toluidine blue O