Structural Features and Anti-coagulant Activity of the Sulphated Polysaccharide SPS-CF from a Green Alga Capsosiphon fulvescens.
Previously, we reported that the sulphated polysaccharides (SPS)-CF, a water-soluble polysaccharide isolated and purified from Korean green alga Maesaengi (Capsosiphon fulvescens, Chlorophyta), is a glucuronogalactomannan based mainly on the monosaccharide composition determined by high-performance liquid chromatography (HPLC) analysis after 1-phenyl-3-methyl-5-pyrazolone (PMP) labelling of sugars in the acid (trifluoroacetic acid (TFA)) hydrolyzates of SPS-CF, which showed mannose (55.4 mol %), galactose (25.3 mol %) and glucuronic acid (16.3 mol %) as major sugars (Na et al., Int Immunopharmacol 10:364-370, 2010). However, the results of the present study re-performed for monosaccharide composition of this polysaccharide using, in addition to HPLC of PMP-labelled sugars, other separation methods, i.e. high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD), gas chromatography with flame ionising detection (GC-FID) and thin-layer chromatography (TLC), clearly demonstrated that the most prominent neutral monosaccharides of SPS-CF are xylose (38.6-49.4 mol %) and rhamnose (39.6-45 mol %), while mannose and galactose are present at a much lesser extent or in negligible amount. These extensive monosaccharide analyses, correlation nuclear magnetic resonance (NMR), electrospray ionization mass spectrometry (ESI-MS) and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) measurements confirmed the sulphated glucuronorhamnoxylan (ulvan) type of SPS-CF polysaccharide, whose backbone is composed of alternating sequence of 4-linked L-rhamnose-3-sulphate and D-xylose residues (ulvobiose U3s) carrying monomeric D-glucuronic acid or D-glucuronic acid-3-sulphate on O-2 of some L-rhamnose-3-sulphate units as the side chains. The SPS-CF exhibited significant in vitro anti-coagulant activity by which the activated partial thromboplastin time (aPTT) and thrombin time (TT) were significantly prolonged. The results of this study demonstrated that the ulvan SPS-CF isolated from Korean Maesaengi C. fulvescens can be considered a potential anti-coagulant agent.
Department of Carbohydrate Chemistry and Technology, Institute of Chemical Technology in Prague, Technická 5, 166 28, Prague 6, Czech Republic.
Andriy
A
Synytsya
Department of Biotechnology and The Research Centre for Biopharmaceutical Lead Molecule (GRRC), The Catholic University of Korea, Bucheon, Gyeonggi-do, 420-743, Republic of Korea.
Doo Jin
DJ
Choi
Institute of Organic Chemistry and Biochemistry AS CR, Flemingovo sq. 2, 166 28, Prague 6, Czech Republic.
Radek
R
Pohl
Department of Biotechnology and The Research Centre for Biopharmaceutical Lead Molecule (GRRC), The Catholic University of Korea, Bucheon, Gyeonggi-do, 420-743, Republic of Korea.
Ye Seul
YS
Na
Institute of Chemistry, Centre for Glycomics, Slovak Academy of Sciences, Dúbravská cesta 9, 845 38, Bratislava, Slovakia.
Peter
P
Capek
Institute of Chemistry, Centre for Glycomics, Slovak Academy of Sciences, Dúbravská cesta 9, 845 38, Bratislava, Slovakia.
Erika
E
Lattová
Department of Carbohydrate Chemistry and Technology, Institute of Chemical Technology in Prague, Technická 5, 166 28, Prague 6, Czech Republic.
Tomáš
T
Taubner
Department of Biotechnology and The Research Centre for Biopharmaceutical Lead Molecule (GRRC), The Catholic University of Korea, Bucheon, Gyeonggi-do, 420-743, Republic of Korea.
Ji Won
JW
Choi
Department of Biotechnology and The Research Centre for Biopharmaceutical Lead Molecule (GRRC), The Catholic University of Korea, Bucheon, Gyeonggi-do, 420-743, Republic of Korea.
Chang Won
CW
Lee
Department of Pharmaceutical Science, Gachon University, Yeonsu-gu, Incheon, 406-799, Republic of Korea.
Jae Kweon
JK
Park
Department of Biotechnology and The Research Centre for Biopharmaceutical Lead Molecule (GRRC), The Catholic University of Korea, Bucheon, Gyeonggi-do, 420-743, Republic of Korea.
Woo Jung
WJ
Kim
Department of Biotechnology and The Research Centre for Biopharmaceutical Lead Molecule (GRRC), The Catholic University of Korea, Bucheon, Gyeonggi-do, 420-743, Republic of Korea.
Sung Min
SM
Kim
Department of Biotechnology and The Research Centre for Biopharmaceutical Lead Molecule (GRRC), The Catholic University of Korea, Bucheon, Gyeonggi-do, 420-743, Republic of Korea.
Jisun
J
Lee
Department of Biotechnology and The Research Centre for Biopharmaceutical Lead Molecule (GRRC), The Catholic University of Korea, Bucheon, Gyeonggi-do, 420-743, Republic of Korea. yongil382@catholic.ac.kr.
Yong Il
YI
Park
D000925:Anticoagulants / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D000460:Chlorophyta / Q000378:metabolism*; D002849:Chromatography, Gas; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D005410:Flame Ionization; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008351:Mannans / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D009005:Monosaccharides / Q000032:analysis; D010314:Partial Thromboplastin Time; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013918:Thrombin Time
D000925:Anticoagulants; D008351:Mannans; D009005:Monosaccharides
Alga Maesaengi (Capsosiphon fulvescens); Anti-coagulant activity; Monosaccharide composition; Structure; Ulvan