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Previous issue date: 2011-09-16 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior, CAPES. / CRUZ, Wellington Oliveira. Characterization of Alphitobius diaperinus (Coleoptera,
Tenebrionidae) ?-amylase: Effect of Different Birds Diet and Inhibitors from Phaseolus
vulgaris. 2011. 50 p Disserta??o (Mestrado em Qu?mica, ?rea de Concentra??o Bioqu?mica).
Instituto de Ci?ncias Exatas, Departamento de Qu?mica, Universidade Federal Rural do Rio
de Janeiro, Serop?dica, RJ, 2011.
Alphitobius diaperinus known as mealworm of poultry litter, is one of the main
poultry pests. It uses high protein and carbohydrates substrate of birds as a food source,
colonizing all the production area finding a favorable environment for its proliferation. This
insect causes sanitary and economic problems, affecting not only healthy and growth of the
poultry but also acting as a microorganisms transmitter like bacterias, protozoas and virus. In
this work it was identified an ?-amylase activity in crude extracts of A. diaperinus larvae and
adults. This enzyme is responsible for starch degradation used as a energy source extremely
important for survival of many insects. The ?-amylase was partially purified by ammonium
sulfate precipitation (25-75%) in a chromatographic Sephadex column G (50-80),
polyacrylamide gel revealed only one band with molecular weight of approximately 30 kDa,
its purity was confirmed by the method of enzyme activity assay dinitrosalicylic 3.5-alkaline
(DNS) was increased 7.2 times when compared with the crude extract. The ?-amylase assays
presented a maximum activity at a pH 5.0 and pH 5.6 and an optimal activity temperature of
50 ? C. The maximum activity of ?-amylase A. diaperinus was achieved in the presence of
0.02 mM CaCl2, and inhibited by increasing this reaction concentration. The enzymatic
activity of A. diaperinus ?-amylase was significantly inhibited by amylase inhibitor ?-AIs
(lectin-like inhibitor) of Phaseolus vulgaris, suggesting that this inhibitors can be used as an
important tool in biological control and development of these pests by reducing the digestion
reducing amylase activity which plays an important role in the starch digestion / CRUZ, Wellington Oliveira. Caracteriza??o de ?-amilase de Alphitobius diaperinus
(Coleoptera, Tenebrionidae): Efeito de Diferentes Ra??es de Aves e dos Inibidores de
Phaseolus Vulgaris. 2011. 50 p Disserta??o (Mestrado em Qu?mica, ?rea de Concentra??o
Bioqu?mica). Instituto de Ci?ncias Exatas, Departamento de Qu?mica, Universidade Federal
Rural do Rio de Janeiro, Serop?dica, RJ, 2011.
Alphitobius diaperinus, conhecido como cascudinho de cama avi?ria, ? uma das
principais pragas av?colas. Ele utiliza o substrato das aves rico em prote?nas e carboidratos
como fonte de alimento, colonizando toda a ?rea de produ??o, encontrando um ambiente
favor?vel para sua prolifera??o. Este inseto causa problemas sanit?rios e econ?micos,
afetando a sa?de e o crescimento das aves e atuando tamb?m como transmissor de
microrganismos tais como baact?rias, protozo?rios e v?rus. Neste trabalho foi identificada a
presen?a de uma ?-amilase no extrato bruto de larvas e adultos de Alphitobius diaperinus.
Esta enzima digestiva ? respons?vel pela degrada??o de amido utilizado como combust?vel na
obten??o de energia, sendo extremamente importante para sobreviv?ncia de muitos insetos. A
?-amilase foi parcialmente purificada por precipita??o em sulfato de am?nio (25-75 %) em
coluna cromatogr?fica Sephadex G (50-80), revelando em gel de poliacrilamida uma ?nica
banda com peso molecular de aproximadamente 30 KDa. Sua pureza enzim?tica foi
confirmada atrav?s de ensaio de atividade pelo m?todo 3,5-dinitrosalic?lico alcalino (DNS),
observando-se um aumento de 7.2 vezes em rela??o ao extrato bruto. A ?-amilase ensaiada
apresentou uma atividade m?xima em pH 5.0 e pH 5.6 e uma temperatura de atividade ?tima
de 50?C. A atividade m?xima da ?-amilase de A. diaperinus foi alcan?ada na presen?a de 0.02
mM de CaCl2, e inibida com o aumento da concentra??o deste reagente. A atividade
enzim?tica da ?-amilase de A. diaperinus, foi significativamente inibida pelo inibidor ?-AIs,
(inibidores tipo lectina) de Phaseolus vulgaris, sugerindo que estes podem ser utilizados
como importantes ferramentas biol?gicas no controle e desenvolvimento de A. diaperinus
atrav?s da redu??o da atividade digestiva destas pragas devido a atua??o desses inibidores
sobre a a??o catalisadora de amilases, enzimas fundamentais para a digest?o de amido
| Identifer | oai:union.ndltd.org:IBICT/oai:localhost:jspui/1145 |
| Date | 16 September 2011 |
| Creators | Cruz, Wellington Oliveira da |
| Contributors | Pontes, Emerson Guedes |
| Publisher | Universidade Federal Rural do Rio de Janeiro, Programa de P?s-Gradua??o em Qu?mica, UFRRJ, Brasil, Instituto de Ci?ncias Exatas |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UFRRJ, instname:Universidade Federal Rural do Rio de Janeiro, instacron:UFRRJ |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | 7 REFER? CIAS BIBLIOGR?FICAS ARENDS, J.J. Control, management of the litter beetle. Poultry Digest, v.44, p.172- 176, (1987). ALKAZAZ, M.; DESSEAUX, V.; MARCHIS-MOUREN, G.;PAYAN, F.; FOREST, E.; SANTIMONE, M. The mechanism of porcine pancreatic alpha-amylase. Kinetic evidence for two additional carbohydrate-binding sites. European Journal Biochemistry, 241, 787-796, (1996). AXTELL, R. C. Department of Entomology, orth Carolina State University, Raleigh, NC 2., p. 7695-7613, (1999). BAKER, J. E. Properties of amylases from midguts of larvae of Sitophilus zeamais and Sitophilus granaries. Insect Biochemistry, 13: p.421-428, (1983). BERNFELD, P. Amylase ? and ?. Methods in Enzymology 1 p.149?158, (1955). BOOTH, R. G.; COX , M. L.; MADGE , R. B. II Guides to insects of importance to man. 3. Coleoptcra. Londres, International journal Entomology: 384p, (1990). BO?I?, N.; VUJ?I?, Z.; NENADOVI?, V.; IVANOVI?, J. Partial purification and characterization of midgut leucyl aminopeptidase of Morimus funereus (Coleoptera: Cerambycidae) larvae. Comparative Biochemistry and Physiology: 134B, p. 231? 234, (2003). BUONOCORE, V.; POERIO, E.; PACE, W.; PETRUCCI, T.; SILANO, V.; TOMASI, M. Interaction of Tenebrio molitor L. ?-amylase with wheat our protein inhibitor. FEBS Letters: 67, p. 202 - 206, (1976). C. BOMPARD-GILLES.; P. ROUSSEAU.; P. ROUGE.; F. PAYAN. Substrate mimicry in the active center of a mammalian alpha-amylase: structural analysis of an enzyme? inhibitor complex, Structure: v.4, p.1441? 1452, (1996). CAMPOS, F.A.P.; XAVIER-FILHO, J.; SILVA, C. P.; ARY, M. B. Resolution and partial characterization of proteinases and ?-amylases from midguts of larvae of the Bruchid beetle Callosobruchos maculatus. Comparative Biochemistry and Physiology: 92B, p. 51-57, (1989). CHERNAKI, A. M.; ALMEIDA, L. M. Morfologia dos est?gios imaturos e do adulto de Alphitobius diaperinus (Panzer) (Coleoptera, Tenebrionidae). Revista brasileira de Zoologia: V.18 (2): p. 351 ? 363, (2001). CHERNAKI, A. M.; ALMEIDA, L. M. Exig?ncias T?rmicas, Per?odo de Desenvolvimento e Sobreviv?ncia de Imaturos de Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae). eotropical Entomology: 30 (3) p.365-368, (2001). 46 CHERNAKI-LEFFER, A.M.; LAZZARI, F. A.; LAZZARI, S. M. N.; ALMEIDA, L. M. Controle do cascudinho. Avicultura Industrial: 1094: p. 22-25, (2001). CHIPOULET, J.M.; CHARARAS, C. Survey and Electrophoretical Separation of the Glycosidases of Rhagium inquisitor (Coleoptera: Cerambycidae) larvae. Comparative Biochemistry and Physiology: 80B: p. 241?246, (1985). CRISTOFOLETTI, P.T.; RIBEIRO, A.F.; TERRA, W. R. Apocrine secretion of amylase and exocytosis of trypsin along the midgut of Tenebrio molitor larvae. Journal of Insect Physiology: 47: p. 143-155, (2001). DE LAS CASAS, E.; POMEROY, B. S.; HAREIN, P. K. Infection and quantitative recovery of Salmonella Typhimurium and Escherichia Coli from within the lesser mealworm, Alphitobius Diaperinus (Panzer). Poultry Science: 47(6): p. 1871-5, (1968). DOJNOV, B. Purification and properties of midgut ?-amilase isolated form from Morimus funereus larvae. Comparative Biochemistry and Physiology, 149, p.153- 160, (2008). DUNFORD, J. C.; KAUFMAN, P. E. Lesser Mealworm, Litter Beetle, Alphitobius diaperinus (Panzer) (Insecta: Coleoptera): Service, Institute of Food and Agricultural Sciences: University of Florida. Published March, Revised: June (2006). EIDSON, C.S.; SCHMITTLE, S. C.; LAL, J. B.; GOODE, R. B. The role of darkling beetle (Alphitobius diaperinus) in the transmission of acute leukosis in chickens. Poultry Science : 44: p. 1366-1367, (1965). F.M. LAJOLO.; F. FINARDO FILHO. Partial characterization of the amylase inhibitor of black beans (Phaseolus vulgaris), variety Rico 23, Journal of Agricutural Food Chemistry: 33, p.132?138, (1985). FALOMO, A. The Pheromone Biology of the Lesser Mealworm Alphitobius diaperinus (Panzer), (Coleoptera: Tenebrionidae). Thesis, University of Wisconsin-Madison: (1986). FENG, G. H.; RICHARDSON, M.; CHEN, M. S.; KRAMER, K. J.; MORGAN, T. D.; AND REECK, G. R. ?-amylase inhibitors from wheat: sequences and patterns of inhibition of insects and human ?-amylases. Insect Biochemistry: Mol. Biol.26 (5): p. 419?426, (1996). FINARDI-FILHO, E.; MIRKOV, M. J.; CHRISPEELS, A. Putative precursor protein in the evolution of the bean ?-amylase inhibitor. Phitochemistry: 43, p. 57-62, (1996). FRANCO, O. L.; RIGDEN, D. J.; MELO, F. R.; BLOCH J. R. C.; SILVA, C. P.; GROSSI DE S?, M. F. Activity of wheat ?- amylase inhibitors towards bruchid ?- amylases and structural explanation of observed specificities. European Journal Biochemistry: 267 (8), p. 1466-1473, (2000). 47 FRANCO, O. L.; RIGDEN, D. J.; MELO, F. R.; GROSSI DE S?, M. F. Plant ?- amylase inhibitors and their interaction with insect ?-amylases, European Journal Biochemistry: 269, p. 397?412, (2002). FRANCO. O. L.; MELO, F. R.; MENDES, P. A.; PAES, N. S.; YOKOYAMA, M.; COUTINHO, M. V.; BLOCH JR, C.; GROSSI DE S?. Characterization of two Acanthoscelides obtectus ?-amylases and their inactivation by wheat inhibitors, J. Journal of Agricutural Food Chemistry: 53, p. 1585?1590, (2005). FRELS, J. M.; RUPNOW, J. H. Purification and partial characterization of two ?- amylase inhibitors from black bean (Phaseolus Vulgaris). Jounal of Agricutural Food Chemistry: 8, p. 281-301,(1984). GIRI, P. A.; KACHOLE, S. M. Detection of electrophoretically separated amylase inhibitors in starch ? polyacrylamide gels. Journal of Chromatography A: 752, p. 261-264, (1996). GOODWIN, M. A.; WALTMAN, W. D. Transmission of Eimeria, viruses and bacteria to chicks: darkling beetles Alphitobius diaperinus as vector of pathogens. Journal of Applied Poultry Research: 5, p. 51-55, (1996). GROSSI DE S?, M. F.; MIRKOV, T. E.; ISHIMOTO, M.; COLUCCI, G.; BATEMAN, K. S.; CHRISPEELS, M. J. Molecular characterization of a bean ?- amylase inhibitor that inhibits the ?-amylase of the Mexican bean weevil Zabrotes subfasciatus. Planta: 203, p. 295?303, (1997). HO, M. F.; WHITAKER, J. R. Purification and partial characterization of white kidney beans (Phaseolus Vulgaris) ?-amylase inhibitors from two experimental cultivars. Journal Food Biochemistry: 17, p. 15 ? 33, (1993). HO, M. F.; YIN, X.; FILHO, F. F.; LAJOLO, F.; WHITAKER, J. R. Naturally occurring ?-amylase inhibitors: Structure/function relationships. In Protein Structure- Function Relationships in Foods; Yada, R. Y.; Jackman, R. L.; Smith, J. L.; Eds.; Bishopbriggs, Blackie, Academic and Professional: Glasgow, Scotland, pp 89-119, (1994). HONAVAR, P. M.; SHIH, C. V.; LIENER, I. E. Inhibition of the Growth of Rats by Purified Hemaglutinin Fractions Isolated from Phaseolus vulgaris. Department of Agricultural Biochemistry, University of Minnesota,St. Paul, Minnesota, Journal utrition: 77, p. 109-114, (1962). ISHIMOTO, M.; CHRISPEELS, M. J. Protective mechanisms of the Mexican bean weevil against high levels of ?-amylase inhibitor in the common bean, Phaseolus vulgaris. Plant Physiology: 111, p. 393-401, (1996). JANECEK, S. Structural Features and Evolutionary Relationships in the ?-Amylase Family. In: Glycoenzymes (Eds. M. Ohnishi, T. Hayashi, S. Ishijima, T. Kuriki), pp. 19-54, Japan Scientific Societies Press: Tokyo, (2000). 48 KASAHARA, K.; HAYASHI, K.; ARAKAWA, T.; PHILO, J. S.; WEN, J.; HARA, S.; YAMAGUCHI, H. Complete sequence, subunit structure, and complexes with pancreatic ?-amylase of an ?-amylase inhibitor from Phaseolus vulgaris white kidney beans, Journal Biochemistry: 120, p. 177?183, (1996). KRAULIS, P .J. Molscript: a program to produce both detailed and schematic plots of protein structures. Journal of Applied Crystallography: 24, p. 946 ? 950, (1991). LAEMMLI, U. K.; Cleavage of structural proteins during the assembly of the bacteriophage T4. ature: 227, p. 680?685, (1970). LEMOS, F. J. A.; CAMPOS, F. A. P.; SILVA, C. P.; XAVIER-FILHO, J. Proteinases and amylases of larval midgut of Zabrotes subfasciatus reared on cowpea (Vigna unguiculata) seeds. Entomologia Experimentalis et Applicata: 56, 219?227, (1990). LESCHEN, R. A. B.; STEELMAN, C. D. Alphitobius diaperinus (Coleoptera: Tenebrionidae) larve and adult mouthparts. Entomological ews: v. 99, n. 4, p. 221- 224, (1988). LOMBRA?A, M.; SU?REZ, P.; SAN JUAN, F. Two forms of ?-amylase in mantle tissue of Mytilus galloprovincialis: purification and molecular properties of form II. Comparative Biochemistry Physiology: 142B, 56?66, (2005). LOM?NACO, C.; PRADO, A. P. Estrutura comunit?ria e din?mica populacional da fauna de d?pteros e seus inimigos naturais em granjas av?colas. Anais da Sociedade Entomol?gica do Brasil: 23 (I), p 71-80, (1994). LOWRY O. H.; ROSEBROUGH N. J.; FARR A. L.; RANDALL R. J. Protein measurement with the Folin phenol reagent - Journal Biological Chemistry: 265-275, (1951). MARSHALL, J. J.; LAUDA, C. M. Purification and properties of phaseolamin, an inhibitor of ?-amylase, from the kidney bean, Phaseolus vulgaris. Journal Biological Chemistry: 250, p. 8030-8037, (1975). MORENO, J.; CHRISPEELS, M. J. A lectin gene encodes the ?-amylase inhibitor of the common bean. Proceedings of the ational Academy of Sciences: USA 86, p. 7885-7889, (1989). NODAN, R O.; TSAI, S. M.; GILBERTEON, R. L.; GEPTS, P. Towards an integrated linkage map of common bean 2. Development of an RFLP-based linkage map. Theor. Appl. Genet: 85, p. 513-520, (1993). PAYAN, F. Structural basis for the inhibition of mammalian and insect ?-amylases by plant protein inhibitors, Biochimica et Biophysica Acta: 1696, p. 171-180, (2004). PODOLER, H.; APPLEBAUM, S.W.; The ?-amylase of the beetle Callosobruchus chinensis. Biochemistry Faculty of Agriculture: The Hebrew University, Rehovot, Israel, 121, p. 321?325, (1971). 49 PREISS, F. J.; DAVIDSON, J. A. Characters for Separating Late-Stage Larvae, Pupae, and Adults of Alphitobius diaperinus and A. laevigatus (Coleoptera: Tenebrionidae) Source: Annals of the Entomological Society of America: Volume 63, Number 3, 15, pp. 807-808, May (1970). PUEYO, J. J.; HUNT, D. C.; CHRISPEELS, M. J. Activation of bean (Phaseolus vulgaris) ?-amylase inhibitor requires proteolytic processing of the pro-protein. Plant Physiology: 101, p. 1341-1348, (1993). RICHARDSON, M. Seed storage protein: the enzyme inhibitor, In: Methods in Plant Biochemistr: New York: Acad Press, v. 5 p. 295 ? 305, (1991). ROUGE, P.; BARRE, A.; CAUSSE, H.; CHATELAIN, C.; PORTHE, G. Arcelin and ?-amylase inhibitor from the seeds of common bean (Phaseolus Vulgaris ) are truncated lectins. Biochemical Systematics and Ecology: 21, p. 695-703, (1993). SARIN, K. Digestive enzymes in immature and adult stages of Alphitobius diaperinus. Entomological society of Japan: 41, p. 10-17, (1973). SILVA C. P.; TERRA W. R.; XAVIER-FILHO, J.; GROSSI DE S?, M. F.; LOPES, A. R.; PONTES, E. G. Digestion in larvae of Callosobruchus maculatus and Zabrotes subfasciatus (Coleoptera: Bruchidae) with emphasis on ?-amylases and oligosaccharidases. Insect Biochemistry and Molecular Biology: 29, p. 355?366, (1999). SILVA D. P.; CASADO-FILHO, E. L.; CORREA, A. S.; FARIAS L. R.; BLOCH J. R. C.; GROSSI-DE-S? M. F.; MENDES P. A.; QUIRNO B. F.; NORONHA E. F.; FRANCO O. L.; Identification of an ?-amylase inhibitor from Pterodon pubescens with ability to inhibit cowpea weevil digestive enzymes, Journal Agricultural Food Chemistry: 55,4382?4387, 2007. SILVA, C. P.; TERRA, W. R.; XAVIER-FILHO, J.; GROSSI-DE-S?, M. F.; ISEJIMA, E. M.; DAMATTA, R. A.; MIGUENS, F. C.; BIFANO, T. D.; Digestion of legume starch granules by larvae of Zabrotes subfasciatus (Coleoptera: Bruchidae) and the induction of ?- mylases in response to different diets. Insect Biochemistry and Insect Molecular Biology: 31, p. 41?50, (2001). SIVAKUMAR, S.; MOHAN, M.; FRANCO, O. L.; THAYUMANAVAN, B. Inhibition of insect pest ?-amilase by little and finger millet inhibitors. Pesticide Biochemistry and Physiology: v. 85, n. 3, p. 155-160,(2006). SNEDEKER, C. F. K.; MOULTHROP, I. M. Some studies on the infectious bursal agent. Avian diseases: n. 11, p.519-528, (1967). STROBL, S.; MASKOS, K.; BETZ, M.; WIEGAND, G.; HUBER, R.; GOMIS-RU? TH, F. X.; GLOCKSHUBER, R. Crystal structure of yellow mealworm ?-amylase at 1.64 A ? resolution. Journal of Molecular Biology: 278, p. 617-628, (1998). 50 TERRA, W. R.; FERREIRA, C. Insect digestive enzyme: properties, compartmentalization and function. Comparative Biochemistry and Physiology: Part B 109: p. 1-62, (1994). TERRA, W. R.; FERREIRA, C.; JORD?O, B. P.; DILLON, R. J. Digestive enzymes. In: Biology of the Insect Midgut. Edited by Lehane, M. J.; Billingsley, P. F. Chapman and Hall: London, pp. 153?194, (1996). VALLEE, B. L.; STEIN, E. A.; SUMERWELL, W. N.; FISCHER, E. H. Metal content of ?-amylase of various origins. Journal Biological Chemistry: 234, p. 2901?2905, (1959). VIOLET, M.; MEUNIER, J. C. Kinetic study of the irreversible thermal denaturation of Bacillus licheniformis ?-amylase. Biochemical journal: v. 263, p. 665-670, (1989). WILSON, T. H.; MINNER, F. D. Influence of temperature on development of the lesser mealworm Alphitobius diaperinus. Journal of Kansas Entomology Society: 42, p. 294-303, (1968). YOUNG, N. M.; THIBAULT, P.; WATSON, D. C; CHRISPEELS, M. J. Posttranslational processing of two ?-amylase inhibitors and an arcelin from the common bean, Phaseolus vulgaris. FEBS Letters: 446, p. 203-206, (1999). ZIEGLER, P. Partial purification and characterization of the major endo amylase of mature pea leaves. Plant Physiology: 86, p. 659?666, (1988). |
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