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http://dx.doi.org/10.3746/jkfn.2004.33.8.1262

Purification and Characterization of an Anticoagulant from Corn Silk  

Choi, Sang-Kyu (Dept. of Biological Sciences and Innunonodulation Research Center, University of Ulsan)
Choi, Hye-Seon (Dept. of Biological Sciences and Innunonodulation Research Center, University of Ulsan)
Publication Information
Journal of the Korean Society of Food Science and Nutrition / v.33, no.8, 2004 , pp. 1262-1267 More about this Journal
Abstract
An anticoagulant was purified from corn silk which has been used in Oriental Medicine. The anticoagulant from corn silk has a molecular mass of 135 kDa, and purified by 24 folds with a recovery of 11%. It was not sensitive to heat and protease treatment. However, periodate oxidation of the anticoagulant resulted in loss of activity significantly, implying that a carbohydrate was responsible for an anticoagulant activity. Galactose, glucose, mannose, fucose, glucosamine, and galactosamine were detected after acid hydrolysis by thin layer chromatography (TLC) and Bio-LC. It was confirmed that anticoagulant had OH and NH bonds by IR, supporting that the anticoagulant is composed of neutrosugar and aminosugar. Its anticoagulating activity was measured by delay in thrombin time (TT) and prothrombin time (PT) without affecting clotting by snake venom and delay in activated partial thromboplastin time (APTT). TT was more sensitive than PT, and was delayed two and three times at the concentration of 60 and 88 nM, respectively. The anticoagulating activity was reduced in the thrombin-induced clotting assay using purified fibrinogen according to the increase of fibrinogen concentration with the apparent Ki value of 23 nM.
Keywords
corn silk; anticoagulant; thrombin time; thrombin;
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1 Olson ST, Bjo I, Sheffer R, Craig PA, Shore JD, Choay J. 1992. Role of the antithrombin-binding pentasaccharides in heparin accerleration of antithrombin-proteinase reactions. J Biol Chem 267: 12528-12538
2 Strube KH, Kroger B, Bialojan S, Otte M, Dodt J. 1993. Isolation, sequence anlysis, and cloning of haemadin. J Biol Chem 268: 8590-8595
3 Friedrich T, Kroger B, Bialojan S, Lemaire HG, Hoffken HW, Reuschenbach P, Otte M, Dodt J. 1993. A kazal-type inhibitor with thrombin specificity from Rhodnius prolixus. J Biol Chem 268: 16216-16222
4 Noeske-Jungblut C, Haendler B, Donner P, Alagon A, Possani L, Schleuning WD. 1995. Triabin, a highly potent exosite inhibitor of thrombin. J Biol Chem 270: 28629-28634   DOI
5 Dahlback B, Stenflo J. 1993. A natural anticoagulant pathway: biochemistry and physiology of protein C, S, C4b- binding protein and thrombomodulin. In Haemostasis and thrombosis. 3rd ed. Bloom AL, Forbes CD, Thomas DP, eds. Churchill livingstone, London. p 671-698
6 Jakubowski JA, Maraganore JM. 1990. Inhibition of coagulation and thrombin-induced platelet activities by a synthetic dodecapeptide modeled on the carboxy-terminus of hirudin. Blood 75: 399-406
7 Tsiang M, Lentz SR, Dittman WA, Scarpati EM, Sadler JE. 1990. Equilibrium binding of thrombin to recombinant human thrombomodulin: Effects of hirudin, fibrinogen, factor Va, and peptide analogues. Biochemistry 29: 10602- 10612   DOI   ScienceOn
8 과학백과사전출판사 엮음. 1991. 약초의 성분과 이용. 일월서각, 서울. p 187-189, 519, 651-652
9 전국한의과대학교수공편. 1991. 본초학. 영림사, 서울. p 217-419
10 고경식, 김윤식. 1989. 한국원색식물도감. 도서출판 아카데미서적, 서울. p 264
11 Sa YS, Kim KA, Choi HS. 2003. Purification and characterization of anti-coagulant activity from persimmon stem. J Korean Soc Foods Sci Nutr 32: 1323-1327   DOI   ScienceOn
12 Yun SI, Cho HR, Choi HS. 2002. Anticoagulant from Taraxacum platycarpum. Biosci Biotechnol Biochem 66: 1859-1864   DOI   ScienceOn
13 Cho HR, Choi HS. 2003. Effects of anticoagulant from Spirodela polyrhiza in rats. Biosci Biotechnol Biochem 67: 881-883   DOI   ScienceOn
14 Naski MC, Fenton II JW, Maraganore JM, Olson ST, Shafer JA. 1990. The COOH-terminal domain of hirudin. J Biol Chem 265: 13484-13489
15 Herbert JM, Herault JP, Bernat A, van Amsterdam RGM, Lormeau JC, Petitou M, van Boeckel C, Hoffmann P, Meuleman DG. 1998. Biochemical and pharmacological properties of Sanorg 34006, a potent and long-acting synthetic pentasaccharide. Blood 91: 4197-4205
16 Kim W, Choi K, Kim Y, Park H, Choi J, Lee Y, Oh H, Kwon I, Lee S. 1996. Purification and characterization of a fibrinolytic enzyme produced from Bacillus sp. strain CK 11-4 screened from chungkook-jang. Appl Environ Microbiol 62: 2482-2488
17 Sumi H, Hamada H, Nakanishi K, Hiratani H. 1990. Enhancement of the fibrinolytic activity in plasma by oral administration of NK. Acta Haematol 84: 139-143   DOI   ScienceOn
18 Choi HS, Shin HH. 1998. Purification and characterization of a fibrinolytic protease in Pleurotus ostreatus. Mycologia 90: 674-679   DOI   ScienceOn
19 Shin HH, Choi HS. 1999. Purification and partial characterization of a metalloprotease in Flammulina velutipes. J Microbiol 36: 20-25
20 Choi HS, Sa YS. 2000. Fibrinolytic and antithrombotic protease from Ganoderma lucidum Mycologia 92: 545-552   DOI   ScienceOn
21 Choi HS, Sa YS. 2001. Fibrinolytic and antithrombotic protease from Spirodela polyrhiza. Biosci Biotechnol Biochem 65: 781-786   DOI   ScienceOn
22 Stringer HA, Pannekoek H. 1995. The significance of fibrin binding by plasminogen activaotor inhibitor 1 for the mechanism of tissue-type plasminogen activator-mediated fibrinolysis. J Biol Chem 270: 11205-11208   DOI   ScienceOn
23 Sumi H, Hamada H, Tsushima H, Mihara H, Muraki H. 1987. A novel fibrinolytic enzyme in the vegetable cheese Natto: a typical and popular soybean food in the Japanese diet. Experimentia 43: 1110-1111   DOI   ScienceOn
24 Holsat J, Lindblad B, Bergqvist D. 1994. Antithrombotic effect of recombinant truncated tissue factor pathway inhibitor (TFPI1-161) in experimental venous thrombosis-a comparison with low molecular weight heparin. Thromb Haemost 71: 214-219
25 Rezaie AR, Cooper ST, Church FC, Esmon CT. 1995. Protein C inhibitor is a potent inhibitor of the thrombin-thrombomodulin complex. J Biol Chem 270: 25336-25339   DOI   ScienceOn
26 Sherry S. 1987. Recombinant tissue plasminogen activator (rt-PA): is it the thrombolytic agent of choice for an evolving acute myocardial infarction? Am J Cardiol 59: 984-989   DOI   ScienceOn
27 Duval-Jobe C, Parmely MJ. 1994. Regulation of plasminogen activation by human U937 promyelocytic cells. J Biol Chem 269: 21353-21357