Summary
We have found out that cell-free extracts from frozen krill decompose many oligo-and polysacharides, particularly with (1→3)-β-and (1→4)-β-linkages. Two individual proteins have very high activity with laminaran as the substrate. One of them has been isolated and purified 980-fold. Polyacrylamide-gel electrophoresis of purified preparation of krill (1→3)-β-glucanase [(1→3)-β-D-glucan glucanohydrolase, EC 3.2.1.6] demonstrated that it was slightly contaminated by one protein band inactive in laminaran hydrolysis. Studies on the hydrolysis of different substrates showed that the enzyme was able to break down only (1→3)-β-D-linkages by an endo-splitting mechanism. Glucono-δ-lactone and heavy metal ions such as Hg2+ inhibited enzyme activity. The activity of the endo-(1→3)-β-glucanase of krill strictly depended on free thiol groups in a enzyme molecule. The Michaelis constant value for laminaran was 0.063 mg/ml. Optimal determined temperature was 65°C and optimal pH 5.0. Because of this enzyme's strong interaction with concavalin A-Sepharose it is suggested that it might be a glycoprotein.
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References
Ballance GM, Meredith WOS (1976) Purification and partial, characterization of an endo-β-1,3-glucanase from green malt. J Inst Brew, London 82:64–67
Beattie A, Hirst EL, Percival E (1961) Studies on the metabolism of theChrysophyceae. Comparative structural investigation on leucosin (chrysolaminarin) separated from diatoms and laminarin from the brown algae. Biochem J 79:531–537
Clark DR, Johnson J, Chung KM, Kirkwood S (1978) Purification, characterization and action-pattern studies on the endo-(1→3)-β-D-glucanase fromRhizopus arrhizus QM 1032. Carbohydr Res 61:457–477
Davis BJ (1964) Disc electrophoresis. 2. Method and application to serum proteins. Ann NY Acad Sci 121:404–412
Dubois M, Gilles KA, Hamilton HK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356
Farkaš V, Biely P, Bauer Š, (1973) Extracellular β-glucanases of yeastSaccharomyces cerevisiae. Biochim Biophys Acta 321:246–255
Galas E, Turkiewicz M, Kałuzewska M, Antezak T (1979) Initial studies on antarctic krill enzymatic system (in Polish with English summary). In: Studia i Materiały, Seria S, no 1, part I Wydawnictwa Morskiego Instytutu Rybackiego, Gdynia, pp 336–357
Goldstein LJ, Hay GW, Lewis BA, Smith F (1965) Controlled degradation of polysaccharides by periodate oxidation. In: Whistler RL, Wolfram ML (eds) Methods in carbohydrate chemistry, vol V. Academic Press, New York London, pp 361–369
Kobayashi Y, Tanaka H, Ogasawara N (1974) Multiple β-1,3-glucanases in the lytic enzyme complex ofBacillus circulans WL 12. Agric Biol Chem 38:959–965
Lachance MA, Villa TG, Phaff HJ (1977) Purification and partial characterization of an exo-β-glucanase from the yeastKluyveromyces aestuarii. Can J Biochem 55:1001–1006
Lineweaver H, Burk D (1934) The determination of enzyme dissocation constans. J Am Chem Soc 56:658–666
Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Manners DJ, Masson AJ, Patterson JC (1973) The structure of a (1→3)-β-D-glucan from yeast cell walls. Biochem J 135:19–30
Marshall JJ (1974) Studies on the structure and mechanism of action of glycoside hydrolases. 1. Purification and study of some factors affecting the activity ofRhizopus arrhizus (1→3)-β-D-glucanase. Carbohydr Res 34:289–297
Marshall SM, Orr AP (1960) The physiology of crustacea, vol I. Metabolism and growth. Academic Press, New York, p 248
Mauchline J, Fisher LL (1969) The biology of Euphausiids. Adv Mar Biol, vol VII. Academic Press, London New York, p 194
Meredith W von, Schuster K, Narzis L, Kumada J (1967) Über die Gummistoffe der Gerste und ihr Verhalten während der Malz-und Bierberietung. Brauwissenschaft 20:125–135
Moore AF, Stone BA (1972) A beta-1,3-glucan hydrolase fromNicotiana glutinosa. 1. Extraction, purification and physical properties. Biochim Biophys Acta 258:238–247
Nelson N (1957) Arsenomolybdate method of sugar determination. In: Colowick SP, Kaplan NO (eds) Methods enzymol, vol III. Academic Press, New York, pp 85–86
Noguchi A, Janagimoto M, Masakatsu U, Kimura S (1976) Purification and some properties of protease fromEuphausia superba. Nippon Nogei Kagaku Kaishi 50:415–421
Notario V, Villa TG, Villanueva JR (1976) Purification of an exo-β-D-glucanase from cell-free extracts ofCandida utilis. Biochem J 159:555–562
Ornstein L (1964) Disc electrophoresis. 1. Background and theory. Ann NY Acad Sci 121:321–330
Reese ET, Parrish FW (1971) Nojirimycin and D-glucono-1,5-lactone as inhibitors of carbohydratases. Carbohydr Res 18:381–388
Seki N, Sakaya H, Onozawa T (1977) Studies on the proteases of antarctic krill. Nippon Suisan Gakkaishi 43:955–962
Somogyi M (1952) Notes on sugar determination. J Biol Chem 195:19–23
Takahashi M, Komuro M, Soutomi J (1978) Purification and properties of β-1,3-glucanase fromLetinus lepideus. J Ferment Technol 56:499–505
Turkiewicz M, Galas E, Kalinowska H (1982) Microflora of antarctic krill (Euphausia superba). Acta Microbiol Pol 81:175–184
Villa T, Notario V, Villanueva JR (1979) Occurrence of an endo-1,3-β-glucanase in cluture fluids of the yeastCandida utilis. Biochem J 177:107–114
Whelan WJ (1962) Laminaridextrins. Isolation from pachyman. In: Whistler LR, Wolfrom ML (eds) Methods in Carbohydrate Chemistry, vol I. Academic Press, New York London, pp 330–333
Wong YS, MacLahlan GA (1979) 1,3-β-glucanases fromPisum sativum seedlings. Biochim Biophys Acta 571:256–269
Yamato, S, Yadomae T, Miyazami T (1974) Multiplicity of extracellular β-1,3-glucanase ofAlternaria solani. J Ferment Technol 52:706–712
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This work was supported by the Institute of Ecology of Polish Academy of Sciences as a part MR I/29 programme
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Turkiewicz, M., Galas, E. & Zielińska, M. Purification and partial characterization of an endo-(1→3)-β-D-glucanase fromEuphausia superba Dana (Antarctic Krill). Polar Biol 4, 203–211 (1985). https://doi.org/10.1007/BF00999765
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DOI: https://doi.org/10.1007/BF00999765