Purification and characterization of antithrombotics from Syzygium aromaticum (L.) MErr. & PERRY.
Two antithrombotic polysaccharides with relatively high molecular weight (HMW) and low molecular weight (LMW) were isolated from the flower buds of Syzygium aromaticum (L.) MERR. & PERRY (clove) by anion-exchange chromatography, hydrophobic interaction column chromatography and size exclusion chromatography (LMW: EC-2B-IIIa-2, M.W. ca. 34000; HMW: EC-2C-Ia-2, M.W. ca. 103000). The LMW polysaccharide was mainly composed of Rha, Gal, GalA and Ara (molar %: 24.1, 18.9, 18.0 and 17.9, respectively) with 10.8% of sulfate and 18.2% of protein. The HMW fraction consisted of Ara, Gal, Glc and Rha (molar %: 26.0, 23.7, 17.5 and 12.4, respectively) with 15.4% of sulfate and 8.0% of protein. Both polysaccharides had the backbone of type I rhamnogalacturonan and the side chain of arabinan. Also, most of the sulfates were attached at the position 6 of 3-linked galactosyl residues. Compared to the antithrombotic activity of the HMW fraction (plasma clotting time of 145 s in APTT assay), the LMW fraction displayed a slightly low activity (90 s). However, animal studies indicated that crude LMW polysaccharide did not show acute toxicity, while the acute LD50 of the HMW fraction was approximately 2-fold lower than that of heparin.
Lee
J I
JI
Graduate School of Biotechnology, Korea University, Seoul.
Lee
H S
HS
Jun
W J
WJ
Yu
K W
KW
Shin
D H
DH
Hong
B S
BS
Cho
H Y
HY
Yang
H C
HC
D000818:Animals; D000991:Antithrombins / Q000737:chemistry / Q000302:isolation & purification* / Q000633:toxicity; D001780:Blood Coagulation Tests; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D006801:Humans; D019684:Magnoliopsida / Q000737:chemistry*; D051379:Mice; D008970:Molecular Weight; D011134:Polysaccharides / Q000737:chemistry / Q000302:isolation & purification* / Q000633:toxicity
D000991:Antithrombins; D011134:Polysaccharides