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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 |
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