Abstract
Aqueous ethanolic extracts from five species of the genus Eucheuma (Rhodophyta) i.e. E. serra, E. amakusaensis, E. cottonii, E. gelatinae and E. denticulatum, were examined for hemagglutinating activity with vertebrate erythrocytes. All the extracts tested agglutinated trypsin-treated sheep and rabbit erythrocytes as well as untreated sheep erythrocytes. From the extract of E. serra, which exhibited the highest activity, a lectin was purified by precipitation with cold ethanol followed by gel filtration to exhibit a single band on SDS-PAGE. The yield was surprisingly as high as 1000 mg from 100 g powdered alga. The purified lectin was further separated into two isoforms, designated ESA-1(90 mg) and ESA-2 (890 mg), by ion exchange chromatography. Both lectins showed a single protein band with the same molecular weight of 29 000 on SDS-PAGE and differed from each other only in isoelectric point (pI 4.75 for ESA-1 and pI 4.95 for ESA-2). Biochemical studies revealed that both are monomeric proteins without a carbohydrate moiety. The hemagglutinating activities were stable over a wide pH range and at a relatively high temperature. The activities were inhibited by a number of glycoproteins, but not by any of the monosaccharides and disaccharides tested. The lectins showed strong mitogenic activities for mouse lymphocytes.
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Bird KT, Chiles TC, Longley RE, Kendrick AF, Kinkema MD (1993) Agglutinins from marine macro algae of the southeastern United States. J. Appl. Phycol. 5: 213–218.
Chiles TC (1990) Gracilaria tikvahiae agglutinin partial purification and preliminary characterization of its carbohydrate specificity. Carbohydr. Res. 207: 319–326.
Chiles TC, Bird KT (1989) A comparative study of animal erythro-cyte agglutinins from marine algae. Comp. Biochem. Physiol. 94: 107–111.
Goto-Nance R, Muramoto K, Zenpo Y, Kamiya H (1996) Purification and characterization of the lectins of the soft coral Lobophytum variatum. Fish. Sci. 62: 297–301.
Hatakeyama T, Himeshima T, Komatsu A, Yamasaki N (1993) Purification and characterization of two lectins from the sea cucumber Stichopus japonicus. Biosci. Biotech. Biochem. 57: 1736–1739.
Hori K (1994) Marine algal lectins - a new category. Kagaku to Seibutsu 32: 586–594.
Hori K, Ikegami S, Miyazawa K, Ito K (1988) Mitogenic and anti-neoplastic isoagglutinins from the red alga Solieria robusta. Phytochemistry 27: 2063–2067.
Hori K, Matsuda H, Miyazawa K, Ito K (1987) A mitogenic agglu-tinin from the red alga Carpopeltis flabellata. Phytochemistry 26: 1335–1338.
Hori K, Miyazawa K, Fusetani N, Hashimoto K, Ito K (1986a) Hypnins, low-molecular weight peptidic agglutinins isolated from a marine red alga, Hypnea japonica. Biochim. Biophys. Acta 873: 228–236.
Hori K, Miyazawa K, Ito K (1986b) Preliminary characterization of agglutinins from seven marine algal species. Bull. Jap. Soc. Sci. Fish. 52: 323–331.
Hori K, Miyazawa K, Ito K (1990) Some common properties of lectins from marine algae. Hydrobiologia 204/205: 561–566.
Julius MH, Simpson E, Herzenberg LA (1973) A rapid method for the isolation of functional thymus-derived murine lymphocytes. Eur. J. Immunol. 3: 645–648.
Kamiya H, Ogata K, Hori K (1982) Isolation and characterization of a new agglutinin in the red alga Palmaria palmata (L.) O. Kuntze. Bot. mar. 35: 537–540.
Kamiya H, Shiomi K, Shimizu Y (1980) Marine biopolymers with cell specificity - III - agglutinins in the red alga Cystoclonium purpureum: isolation and characterization. J. nat. Prod. 43: 136–139.
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.
Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. immunol. Methods 65: 55–63.
Odo S, Kamino K, Kanai S, Maruyama T, Harayama S (1995) Biochemical characterization of a Ca2+-dependent lectin from the hemolymph of a photosymbiotic marine bivalve, Tridacna derasa. J. Biochem. 117: 965–973.
Rogers DJ, Hori K (1993) Marine algal lectins: new developments. Hydrobiologia 260/261: 589–593.
Sharon N, Lis H (1972) Lectins: cell-agglutinating and sugar-specific proteins. Science 177: 949–959.
Shiomi K, Kamiya H, Shimizu Y (1979) Purification and charac-terization of an agglutinin in the red alga Agardhiella tenera. Biochim. Biophys. Acta 576: 118–127.
Shiomi K, Yamanaka H, Kikuchi T (1981) Purification and physico-chemical properties of hemagglutinin (GVA-1) in the red alga Gracilaria verrucosa. Bull. Jap. Soc. Sci. Fish. 47: 1079–1084.
Tsuboi I, Matsukawa M, Sato N (1993) Isolation and characterization of a sialic acid-specific lectin from hemolymph of the south-east Asian horseshoe crab Tachypleus gigas. Biosci. Biotech. Biochem. 57: 1237–1242.
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Kawakubo, A., Makino, H., Ohnishi, Ji. et al. The marine red alga Eucheuma serra J. Agardh, a high yielding source of two isolectins. Journal of Applied Phycology 9, 331–338 (1997). https://doi.org/10.1023/A:1007915006334
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DOI: https://doi.org/10.1023/A:1007915006334