Anti-coagulant Activity of Isolated and Partially Characterized Proteoglycans and Glycosaminoglycans from African Night Crawler (Eudrilus eugeniae Kinberg)

  • Mia Clare Marie L. Bercansil University of the Philippines Los Baños, College, Laguna 4031
  • Miko Lorenzo J. Belgado University of the Philippines Los Baños, College, Laguna 4031
Keywords: proteoglycan, anti-coagulant, glycosaminoglycans, African night crawler


Proteoglycans and glycosaminoglycans were isolated from African night crawler (Eudrilus eugeniae Kinberg) and partially characterized proteoglycans (3.04 % of lyophilized worm) were liberated from the defatted and depurinated worm samples by dissociative method using 4M urea in acetate buffer. Glycosaminoglycans (12.47% of proteoglycan extract) were extracted using enzymatic hydrolysis of the proteoglycan extract with papain. Gel filtration chromatography using Sepharose CL-4B was used to purify and estimate the molecular weights of the proteoglycan and glycosaminoglycan fractions. Three proteoglycan fractions PGF1, PGF2 and PGF3 with estimated molecular weigths 860 kDa, 181 kDa and 3 kDa, respectively were identified as monitored by the Bradford and modified carbazole assay. Two glycosaminoglycan fractions - GF1 (MW = 860 kDa) and GF2 (MW=140 kDa) were identified using the modified carbazole assay. Infrared spectroscopy of the GF1 and GF2 showed the possible identities of the fractions. GF1 may be a hyaluronic acid and GF2 is possibly chondroitin. Anti-coagulant assay for the extracts and fractions revealed that the glycosaminoglycan isolate has anti-coagulant activity but not the GF1 and GF2 fractions individually.


Athukorala Y, Lee KW, Kim SK, Jeon YJ. Anticoagulant activity of marine green and brown algae collected from Jeju Island in Korea. Bioresource Technol. 2006; 690-756.

Bansil R, Yannas LV, Stanley HE. Raman spectroscopy: A structural probe of glycosaminoglycans. Biochim Biophys Acta. 1978; 541:535-542.

Bradford MM. A dye binding assay for protein. Anal Biochem. 1976; 72:248-254.

Bowness JM. Application of the carbazole reaction to the estimation of glucuronic acid and glucose in some acidic polysaccharides and in urine. Biochem J. 1956; 67:295-300.

Bowness JM. 5-Formylfuroic acid and the carbazole reaction for uronic acids and acidic polysaccharides. Biochem J. 1957; 70:107-110.

Elson LA, Morgan WTJ. A colorimetric method for the determination of glucosamine and chondrosamine. Biochemistry. 1933; 27:1824-1828.

Fraser JRE, Laurent TC, Laurent BG. Hyaluronan: Its nature, distribution, functions and turnover. J Intern Med. 1997; 242:27-33.

Guerrero R. Earthworm culture for vermicompost and vermimeal production and for vermiceutical application in the Philippines (1978-2008) - A Review. Dynamic Soil, Dynamic Plant. Global Science Books; 2009.

Hamed RR, Maharem TM, El-Guindy AS. Proteoglycans from adult worms of Schistosoma haematobium. J Helminthol. 1997; 71:151-160.

Hardingham TE, Fosang AJ. Proteoglycans: many forms and many functions. FASEB J. 1992; 6: 861-870.

Iozzo RV, Ed. Methods of Molecular Biology: Proteoglycan Protocols. Elsevier Scince; 2001.

Karamanos NK, Aletras AJ, Antonopoulos CA, Hjerpe A, Tsiganos CP. Chondroitin proteoglycans from squid skin. Isolation, characterization and immunological studies. Eur J Biochem. 1992; 192:33-38.

Kottamp H, Hindricks G, Breithardt G. 1998. Role of anti-coagulant therapy in atrial fribillation. J Cardiovasc Electro. 1998; 9:86-96.

Nelson DL, Cox MM. Lehninger Principles of Biochemistry. W. H. Freeman; 2005.

Ofosu FA. Mechanisms for the anticoagulant effect of heparin and related polysaccharides. Nouv Rev Fr Matol. 1988; 30(3): 155-160.

Olson SK, Bishop JR, Yates JR, Oegema K, Esko JD. Identification of novelchondroitin proteoglycans in Caenorhobditis elegans: embryonic cell division depends on CPG-1 and CPG-2. J Cell Biol.2006; 173(6):985-994.

Panagos CG, Moss TD, Bavington CD, Olafsson HG, Uhrín D. Characterization of hyaluronic acid and chondrotin/dermatan sulfate from lumpsucker fish C. lumpus. Carbohyd Polym. 2014; 106:25-33.

Swoap OF, Kuzienga MH. 1949. The sheep plasma method for the bioassay of heparin preparations. J Am Pharm Assoc. 1949; 38:563-565.

How to Cite
Bercansil, M. C. M. L., & Belgado, M. L. J. (2015). Anti-coagulant Activity of Isolated and Partially Characterized Proteoglycans and Glycosaminoglycans from African Night Crawler (Eudrilus eugeniae Kinberg). KIMIKA, 26(2), 31-38.
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