Abstract
Agarase, cellulase and alginate lyase activities from crude extracts of Aplysia dactylomela Rang, Haliotis coccinea canariensis Nordsieck, Littorina striata King et Broderip and Diadema antillarum Phillipi were measured in vitro to compare digestive efficiencies against several components of complex seaweed cell walls. Commercial abalone acetone powder (AAP, an extract from Haliotis sp.; Sigma, Ref. A-7514) and purified (Sigma, Ref. A-6306) and non-purified agarases from Pseudomonas atlantica were used with the same objective. Optimum conditions for agarase and cellulase activities were 40°C and pH 6.0. For alginate lyase, optimum temperature and pH were species-dependent. Highest reducing sugar release was shown by crude extracts from A. dactylomela. These crude extracts displayed high agarase activity compared with bacterial agarases at 40°C, and were significantly higher at 25°C. AAP and crude extracts from L. striata and D. antillarum exhibited high specific activities on all the substrates. Cold extract from Gracilaria spp. was the best substrate with which to measure agarase activity.
Similar content being viewed by others
Literature cited
Benítez, L. V., Macaranas, J. M. (1979). Partial purification of a carrageenase from the tropical sea urchin Diadema setosum. Proc. int. Seaweed Symp. 9: 353–359 [Jensen, A., Stein, J. R. (eds.) Science Press, Princeton]
Berliner, M. D. (1981). Protoplasts of eukaryotic algae. Int. Rev. Cytol. 73: 1–19
Björk, M., Ekman, P., Wallin, A., Pedersén, M. (1990). Effects of growth rate and other factors on protoplast yield from four species of Gracilaria (Rhodophyta). Botanica mar. 33: 433–439
Boyen, C., Kloareg, B., Polne-Fuller, M., Gibor, A. (1990). Preparation of alginate lyases from marine molluscs for protoplast isolation in brown algae. Phycologia 29: 173–181
Brock, V., Kennedy, V. S., Brock, A. (1986). Temperature dependency of carbohydrase activity in the hepatopancreas of thirteen estuarine and coastal bivalve species from the North American east coast. J. exp. mar. Biol. Ecol. 103: 87–101
Butler, D. M., Evans, L. V., Kloareg, B. (1990). Isolation of protoplasts from marine macroalgae. In: Akatsuka, I. (ed.) Introduction to applied phycology. SPB Academic Publishing, The Hague, p. 647–668
Butler, D. M., Østgaard, K., Boyen, C., Evans, L. V., Jensen, A., Kloreg, B. (1989). Isolation conditions for high yields of protoplasts from Laminaria saccharina and L. digitata (Phaeophyceae). J. exp. Bot. 40: 1237–1246
Chen, L. C.-M. (1987). Protoplast morphogenesis of Porphyra leucosticta in culture. Botanica mar. 30: 399–403
Cheney, D. P., Mar, E., Saga, N., van der Meer, J. (1986). Protoplast isolation and cell division in the agar-producing seaweed Gracilaria (Rhodophyta). J. Phycol. 22: 238–243
Cocking, E. (1972). Plant cell protoplasts. Isolation and development. A. Rev. Pl. Physiol. 23: 29–50
Davison, I. R., Polne-Fuller, M. (1990). Photosynthesis in protoplasts of Macrocystis pyrifera (Phaeophyta). J. Phycol. 26: 384–387
Duckworth, M., Turvey, J. R. (1969). An extracellular agarase from a Cytophaga species. Biochem. J. 113: 139–142
Elyakova, L. A., Favorov, V. V. (1974). Isolation and certain properties of alginate lyase VI from the mollusk Littorina sp. Biochim. biophys. Acta 358: 341–354
Eppley, R. W., Lasker, R. (1959). Alginase in the sea urchin Strongylocentrotus purpuratus. Science, N.Y. 129: 214–215
Franssen, J., Jeuniaux, C. (1965). Digestion de l'acide alginique chez les invertebres. Cah. Biol. mar. 6: 1–21
Friesen, J. A. (1980). The structure of the gut of Mysis stenolepis and the mechanism of cellulose digestion. M. Sc. thesis. Dalhousie University, Nova Scotia
Galli, D. R., Giese, A. C. (1959). Carbohydrate digestion in a herbivorous snail, Tegula funebralis. J. exp. Zool. 140: 415–439
Horiuchi, S., Lane, C. E. (1965). Digestive enzymes of the crystalline style of Strombus gigas Linne. I. Cellulase and some other carbohydrases. Biol. Bull. mar. biol. Lab., Woods Hole 129: 273–281
Hylleberg Kristensen, J. (1972). Carbohydrases of some marine invertebrates with notes on their food and on the natural occurrence of the carbohydrates studies. Mar. Biol. 14: 130–142
Jordan, P., Kloareg, B., Vilter, H. (1991). Detection of vanadate-dependent bromoperoxidases in protoplasts from the brown algae Laminaria digitata and L. saccharina. J. Pl. Physiol. 137: 520–524
Kidby, D. K., Davidson, D. J. (1973). A convenient ferricyanide estimation of reducing sugars in the nanomole range. Analyt. Biochem. 55: 321–325
Kloareg, B., Quatrano, R. S. (1988). Structure of the cell walls of marine algae and ecophysiological functions of the matrix polysaccharides. Oceanogr. mar. Biol. A. Rev. 26: 259–315
Kreger, D. R. (1962). Cell walls. In: Lewin, R. D. (ed.) Physiology and biochemistry of algae. Academic Press, New York, p. 315–335
Lahaye, M., Rochas, C., Yaphe, W. (1986). A new procedure for determining the heterogeneity of agar polymers in the cell wall of Gracilaria spp. (Gracilariaceae, Rhodophyta). Can. J. Bot. 64: 579–585
Le Gall, Y., Braud, J. P., Kloareg, B. (1990). Protoplast production in Chondrus crispus gametophytes (Gigartinales, Rhodophyta). Pl. Cell Rep. 8: 582–585
Lewis, J. B. (1964). Feeding and digestion in the tropical sea urchin Diadema antillarum Philippi. Can. J. Zool. 42: 549–557
Liu, W. S., Tang, Y. L., Liu, X. W., Fang, T. C. (1984). Studies on the preparation and on the properties of sea snail enzymes. Hydrobiologia 116/117: 319–320
Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. J. biol. Chem. 193: 265–275
Morrice, L. M., McLean, M., Williamson, F. B., Long, W. F. (1983). β-agarases I and II from Pseudomonas atlantica: purifications and some properties. Eur. J. Biochem. 135: 553–558
Morrice, L. M., McLean, M., Williamson, F. B., Long, W. F. (1984). β-agarases from Pseudomonas atlantica. Hydrobiologia 116/117: 576–579
Nakada, H. I., Sweeny, P. C. (1967). Alginic acid degradation by eliminases from abalone hepatopancreas. J. biol. Chem. 242: 845–851
Nelson, N. (1944). A photometric adaptation of the Somogyi method for the determination of glucose. J. biol. Chem. 153: 375–380
Nisizawa, K., Fujibayashi, S., Kashiwabara, Y. (1968). Alginate lyases in the hepatopancreas of a marine mollusc, Dolabella auricula Solander. J. Biochem. 64: 25–37
Onishi, T., Suzuki, M., Kikuchi, R. (1985). The distribution of polysaccharide hydrolase activity in gastropods and bivalves. Bull. Jap. Soc. scient Fish. 51: 301–308
Polne-Fuller, M. (1987). Preparation of enzymes. In: Indergaard, M., Østgaard, K., Guiry, M. D. (eds.) Seaweed protoplast and tissue culture. COST 48. European Economic Community, Brussels. Proceedings of Workshop on Biotechnical Methods in seaweed cultivation. E.E.C., Trondheim, p. 29–30
Polne-Fuller, M., Biniaminov, M., Gibor, A. (1984). Vegetative propagation of Porphyra perforata. Hydrobiologia 116/117: 308–313
Polne-Fuller, M., Gibor, A. (1987a) Microorganisms as digestors of seaweed cell walls. Hydrobiologia 151/152: 405–409
Polne-Fuller, M., Gibor, A. (1987b). Tissue culture of seaweeds. In: Bird, K. T., Benson, P. H. (eds.) Seaweed cultivation for renewable resources. Vol. 16. Elsevier, Amsterdam, p. 219–239
Preston, J. F., Romeo, T., Bromley, J. C., Robinson, R. W., Aldrich, H. C. (1985). Alginate lyase-secreting bacteria Underkofler, L. A. (ed.) Developments in industrial microbiology. Society for Industrial Microbiology, Arlington, Virginia, p. 727–740
Saga, N., Polne-Fuller, M., Gibor, A. (1986). Protoplasts from seaweeds: production and fusion. In: Barclay, W. R., McIntosh, R. P. (eds.) Algal biomass technologies: an interdisciplinary perspective. Vol. 83. J. Cramer, Berlin, p. 37–43
Saga, N., Sakai, Y. (1984). Isolation of protoplasts from Laminaria and Porphyra. Bull. Jap. Soc. scient. Fish. 50: p. 1085
Smith, R. G., Bidwell, R. G. S. (1989). Inorganic carbon uptake by photosynthetically active protoplasts of the red macroalga Chondrus crispus. Mar. Biol. 102: 1–4
Somogyi, M. (1952). Notes on sugar determination. J. biol. Chem. 195: 19–23
Suzuki, M., Watanabe, Y., Tanaka, A., Ohnishi, T. (1986). The polysaccharide degradation activity in the apple snail. Bull. Tokai reg. Fish. Res. Lab. 120: 53–60
Tokuda, H., Kawashima, Y. (1988). Protoplast isolation and culture of a brown alga, Undaria pinnatifida. In: Stadler, T., Mollion, J., Verdus, M.-C., Karamanos, Y., Morran, H., Christiaen, D. (eds.) Algal biotechnology. Elsevier Applied Science, London, p. 152–157
Torzilli, A. P., Andrykovitch, G. (1980). Cell wall degrading enzymes produced by the salt marsh fungus Buergenerula spartinae. Botanica mar. 23: 645–650
Van der Meulen, H. J. (1975). The enzymatic hydrolysis of agar by Cytophaga flevensis sp. nov. Ph. D. thesis. University of Groningen, The Netherlands
Yaphe, W. (1966). The purification and properties of an agarase from a marine bacterium, Pseudomonas atlantica. Proc. 5th int. Seaweed Symp. 5: 333–335 [Young, E. G., McLachlan, J. L. (eds.) Pergamon Press, Oxford]
Yokoe, Y., Yasumasu, I. (1964). The distribution of cellulase in invertebrates. Comp. Biochem. Physiol. 13: 323–338
Zhu, R. (1983). A comparative study of marine algae cell-wall decomposition by the digestive enzymes isolated from three species of marine gastropods. Coll. ocean. Wks Qingdao (China) 6: 122–126
Author information
Authors and Affiliations
Additional information
Communicated by J. M. Pérès, Marseille
Rights and permissions
About this article
Cite this article
Gómez-Pinchetti, J.L., García-Reina, G. Enzymes from marine phycophages that degrade cell walls of seaweeds. Marine Biology 116, 553–558 (1993). https://doi.org/10.1007/BF00355473
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00355473