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Effect of plant protease inhibitors on digestive proteases in two fish species, Lutjanus argentiventris and L. novemfasciatus

Abstract

This work provides a comparative study of the inhibitory effect of several plant protein sources on digestive proteases of two snappers: yellow snapper (Lutjanus argentiventris) and dog snapper (Lutjanus novemfasciatus). Seed extracts did not affect gastric proteases whereas they significantly inhibit intestinal proteases. Inhibition of alkaline proteases showed that pancreatic proteases of L. argentiventris were more sensitive to seed protease inhibitors than those of L. novemfasciatus. Legume seeds showed the highest inhibitory capacity on alkaline proteases causing inhibition higher than 50% in total proteolytic activity. Protease inhibition on digestive extracts was assessed using different relative concentration of seed extracts and represented by constructing dose response curves. In order to reduce the inhibitory effect, seed extracts were acid-treated before the inhibition assay. Results showed that acid treatment did not affect the inhibitory capacity of seeds on alkaline proteases in both species. However, when the action of gastric enzymes was simulated on seed extracts, the inhibitory capacity was reduced significantly, mainly in the case of L. novemfasciatus. The responses of fish enzymes to heat-treated seed extracts were also tested. Only higher temperatures were capable of reducing the inhibitory capacity of seed, with the specific response to the snapper species. The use of biochemical assays allows us to quantify the action of inhibitors on total proteolytic activity. In addition, zymograms obtained by substrate-SDS-PAGE provided qualitative information about the number and type of proteases affected by each inhibitor. Each seed extract produces a characteristic profile of inhibition on alkaline protease. The results obtained are important for future formulation of feeds for these snapper species.

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References

  • Alarcón, F.J., Díaz, M., Moyano, F.J. and Abellán, E. 1998. Characterization and funtional properties of digestive proteases in two sparids; gilthead sea bream (Sparus aurata) and common dentex (Dentex dentex). Fish Physiol. Biochem. 19: 257–267.

    Google Scholar 

  • Alarcón, F.J., Moyano, F.J. and Díaz, M. 1999. Effect of inhibitors present in protein sources on digestive proteases of juvenile sea bream (Sparus aurata). Aquatic Living Res. 12(4): 233–238.

    Google Scholar 

  • Bradford, M.M. 1976. A rapid and sensitive method for the quantifi-cation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248–254.

    Google Scholar 

  • Broadway, R.E. 1995. Are Insects resistant to plant proteinase inhibitors? J. Insect Physiol. 41(2): 107–116.

    Google Scholar 

  • De Silva, S.S. and Anderson, T.A. 1995. Fish Nutrition in Aquaculture, 1st ed. Chapman and Hall, London.

    Google Scholar 

  • Diamond, J. 1997. Guns, Germs, and Steel: The Fates of Human Societies. W.W. Norton and Company, USA.

    Google Scholar 

  • El-Dahhar, A.A. 1999. Effect of heat-treated feed and exogenous zymogen on survival and growth of grey mullet, Liza ramada (Riss), larvae in Egypt. Aquaculture Res. 30: 165–173.

    Google Scholar 

  • Fagbenro, O.A. 1999. Formulation and evaluation of diets for the African catfish, Clarias gariepinus (Burchell), made by partial replacement of fish meal with winged bean (Psophocarpus tetragonolobus L. DC) seed meal. Aquaculture Res. 30: 249–257.

    Google Scholar 

  • Flavin, D.F. 1982. The effects of soybean trypsin inhibitors on the pancreas of animals and man: a review. Vet. Hum. Tox. 24: 25–28.

    Google Scholar 

  • Gallaher, D. and Schneeman, B.O. 1984. Nutritional and metabolic response to plant inhibitors of digestive enzymes. In: Nutritional and Toxicological Aspects of Food Safety. pp. 299–320. Edited by M. Friedman. Plenum Press.

  • García-Carreño, F.L., Dimes, L.E. and Haard, N.F. 1993. Substrategel electrophoresis for composition and molecular weight of proteinases and proteinaceous proteinase inhibitors. Anal. Biochem. 214: 65–69.

    Google Scholar 

  • García-Carreño, F.L., Navarrete del Toro, M.A., Díaz-López, M., Hernández-Cortés, M.P. and Ezquerra, J.M. 1996. Protease inhibition of fish muscle enzymes using legume seeds extracts. J. Food Protection 59: 312–318.

    Google Scholar 

  • García-Carreño, F.L., Navarrete del Toro, A. and Ezquerra, M., 1997. Digestive shrimp proteases for evaluation of protein digestibility in vitro. I: effects of protease inhibitors in protein ingredients. J. Mar. Biotechnol. 5: 36–40.

    Google Scholar 

  • García-Olmedo, F., Salcedo, G., Sánchez-Monge, R., Gómez, l., Royo, J. and Carbonero, P. 1987. Plant proteinaceous inhibitors or proteinases and amylases. Oxford Surveys Plant Mol. Cell. Biol. 4: 275–334.

    Google Scholar 

  • Haard N.F., Dimes L.E., Arndt R.E. and Dong F.M. 1996. Estimation of protein digestibility. IV. Digestive proteinases from the pyloric caeca of coho salmon (Oncorhynchus kisutch) fed diets containing soybean meal. Comp. Biochem. Physiol. B 110(4): 533–540.

    Google Scholar 

  • Holm, H. and Krogdahl, A. 1982. Problems in predicting the inhibition of human pancreas proteinaes by soya bean proteinase inhibitors: in vitro assasys employing human, bovine and porcine proteinases. J. Sci. Food Agric. 33: 1164–1171.

    Google Scholar 

  • Krogdahl, A. and Holm H. 1979. Inhibition of human and rat pancreatic proteinases by crude purified soybean proteinase inhibitors. J. Nutr. 109: 551–558.

    Google Scholar 

  • Krogdahl, A. and Holm, H. 1981. Soybean protease inhibitors and human proteolytic enzymes: selective inactivation of inhibitors by treatments with human gastric juice. J. Nutr. 111: 25–31.

    Google Scholar 

  • Krogdahl, A. and Holm, H. 1983. Pancreatic proteinases from man, trout, rat, pig, cow, chicken, mink and fox. Enzyme activities and inhibition by soybean and lima bean proteinase inhibitors. Comp. Biochem. Physiol. 74: 403–409.

    Google Scholar 

  • Krogdahl, A., Lea, T.B. and Olli, J.J. 1994. Soybean protease inhibitors affect intestinal trypsin activities and amino acids digestibilities in rainbow trout (Oncorhynchus mykiss). Comp. Biochem. Physiol. A 107: 215–219.

    Google Scholar 

  • Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227: 680–685.

    Google Scholar 

  • Liener, I.E. 1980. Toxic constituents of plant foodstuffs. Academic Press, Sydney.

    Google Scholar 

  • Lim, C. and Akiyama, D.M. 1992. Full-fat soybean utilization by fish. Asian Fish. Sci. 5: 181–197.

    Google Scholar 

  • Mitchell A.I., Dawson A. and Houlihan D.F. 1993. Trypsin inhibitors in commercial fish food. pp. 219–222. Edited by Kaushik S.J. and Luquet P. Fish Nutrition in Practice, Les Colloques 61, Biarritz, France.

  • Moyano, F.J., Martínez, I., Díaz, M. and Alarcón, F.J. 1999. Inhibition of digestive proteases by vegetable meals in three species (Sparus aurata), tilapia (Oreochromis niloticus) and african sole (Solea senegalensis). Comp. Biochem. Physiol. B 122: 327–332.

    Google Scholar 

  • Munilla-Morán, R. and Saborio-Rey, F. 1996. Digestive enzymes in marine species. I. Proteinases activities in gut from red-fish (Sebastes mentella), seabream (Sparus aurata) and turbot (Scophthalmus maximus). Comp. Biochem. Physiol. B 113(2): 395–402.

    Google Scholar 

  • Olli, J., Hjelmeland, K. and Krogdahl, A. 1994. Soybean trypsin inhibitor in diets for Atlantic salmon (Salmo salar, L): Effects on nutrient digestibilities and in pyloric caeca homogenate and intestinal content. Comp. Biochem. Physiol. A 109: 923–928.

    Google Scholar 

  • Potempa, l., Korzus, E. and Travis, J. 1994. The serpin superfamily of proteinase inhibitors: structure, function and regulation. J. Biol. Chem. 269: 15957–15960.

    Google Scholar 

  • Reeck, G.R., Kramer, K.J., Baker, J.E., Kanost, M.R., Fabrick, J.A. and Behnke, C.A. 1997. Proteinase inhibitors and resistance of transgenic plants to insects. In: Advance in insect control: the role of transgenic plants. pp. 157–183. Edited by N. Carozi, M. Koziel. Taylor and Francis, London.

    Google Scholar 

  • Ryan, C.A. 1979. Proteinase inhibitors. In: Herbivores: their Interactions with Secondary Plant Metabolites, pp. 599–618. Edited by G.A. Rosenthal and D.H. Janzen. Academic Press, New York.

    Google Scholar 

  • Serrano-Pinto, V. and Caraveo-Patiño, J. 1999. Survival of amarillo snapper Lutjanus argentiventris (Peters 1869) at different salinities in captivity. Aquaculture Res. 30: 467–470.

    Google Scholar 

  • Shimeno, S., Seki, S., Masumoto, T. and Hosokawa, H. 1994. Postfeeding changes in digestion and serum constituent in juvenile yellowtail force-fed with raw and heated defatted soybean meals. Nippon Suis. Gakk. 60(1): 95–99.

    Google Scholar 

  • Strukelj, B. 1992. Characterization of aspartic proteinase inhibitors from potato at the gene, cDNA and protein levels. Biol. Chem. Hoppe Seyler 373: 477–482.

    Google Scholar 

  • Tacon, A.G.J. 1997. Fish meal replacers: Review of antinutrients within oilseeds and pulses. A limiting factor for the aquafeed Green Revolution, Cah. Opt. Médit., 22: 153–182.

    Google Scholar 

  • Whitaker, J.R. 1994. Principles of enzymology for the food sciences. 2nd ed. Marcel Dekker, New York.

    Google Scholar 

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Alarcón, F., García-Carreño, F. & Navarrete del Toro, M. Effect of plant protease inhibitors on digestive proteases in two fish species, Lutjanus argentiventris and L. novemfasciatus . Fish Physiology and Biochemistry 24, 179–189 (2001). https://doi.org/10.1023/A:1014079919461

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  • DOI: https://doi.org/10.1023/A:1014079919461

  • digestive enzyme
  • electrophoresis
  • fish
  • Lutjanus argentiventris
  • L. novemfasciatus
  • plant protein
  • proteases
  • protease inhibitor
  • snapper