Made available in DSpace on 2014-12-17T14:03:27Z (GMT). No. of bitstreams: 1
CarlosEMG.pdf: 434119 bytes, checksum: cffd1dddf77ec049b23dadc1bfa89773 (MD5)
Previous issue date: 2004-12-20 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / A proteinaceous trypsin inhibitor was purified from Crotalaria pallida seeds by ammonium sulphate fractionation, affinity chromatography on immobilized Trypsin-Sepharose and TCA precipitation. The trypsin inhibitor, named ITC, had Mr of 32.5 kDa by SDS-PAGE and was composed by two subunits with 27.7 and 5.6 kDa linked by disulphide bridges, a typical characteristic of Kunitz-Inhibitor family. ITC was stable until 50?C, and at 100?C its residual activity was of about 60%. Also, ITC was stable at pHs 2 to 12. The inhibition of trypsin by ITC was non-competitive, with a Ki of 8,8 x 10-7M. ITC inhibits weakly other serine proteinases such as chymotrypsin and elastase. The inhibition of papain (44% of inhibition), a cysteine proteinase was an indicative of the bi-functionality of ITC. In vitro assays against digestive proteinases from several Lepdoptera, Diptera and Coleoptera pests were made. ITC inhibited in 100% digestive enzymes of Ceratitis capitata (fruit fly), Spodoptera frugiperda and Alabama argillacea, the last one being a cotton pest. It also inhibited in 74.4% Callosobruchus maculatus (bean weevil) digestive enzymes, a Coleoptera pest. ITC, when added in artificial diet models, affected weakly the development of C. capitata larvae and it had a WD50 of 2.65% to C. maculatus larvae / Um inibidor de proteinase foi isolado de sementes de Crotalaria pallida por fracionamento com sulfato de am?nio, cromatografia de afinidade em Sepharose com tripsina acoplada e precipita??o com ?cido tricloroac?tico (TCA). O inibidor de tripsina, denominado ITC, possui massa molecular de 32,5 kDa determinada por SDS-PAGE e ? composto por duas subunidades, uma de 25,7 e outra de 5,6 kDa, ligadas por pontes dissulfeto; uma caracter?stica t?pica da fam?lia de inibidores de Kunitz. ITC foi est?vel at? 50?C e a 100?C sua atividade residual foi cerca de 60%. ITC tamb?m foi est?vel em valores de pHs variando de 2 a 12. A inibi??o da tripsina por ITC ? do tipo n?o-competitivo, com um Ki de 8,8 x 10-7M. ITC inibe fracamente outras proteinases ser?nicas como quimotripsina e elastase. A inibi??o da papa?na (44%), uma proteinase ciste?nica, foi indicativa da bifuncionalidade de ITC. Ensaios in vitro contra proteinases digestivas de v?rios insetos praga das ordens Lepidoptera, Diptera e Coleoptera foram realizados. ITC inibiu em cerca de 100% enzimas digestivas de Ceratitis capitata (mosca-das-frutas), Spodoptera frugiperda (lagarta do cartucho do algod?o), e Alabama argillacea (curuquer? do algod?o). ITC tamb?m inibiu em 74,4% enzimas digestivas de Callosobruchus maculatus (broca do feij?o) uma praga da ordem Coleoptera. ITC, quando adicionado a dietas artificiais, afetou fracamente o desenvolvimento de larvas de C. capitata e teve um WD50 de 2,65% contra larvas de C. maculatus
| Identifer | oai:union.ndltd.org:IBICT/oai:repositorio.ufrn.br:123456789/12530 |
| Date | 20 December 2004 |
| Creators | Gomes, Carlos Eduardo Maia |
| Contributors | CPF:29706106391, http://lattes.cnpq.br/7890362793618911, Vidal, M?rcia Soares, CPF:02621094767, http://lattes.cnpq.br/3036544314910366, Braz, Regina de F?tima dos Santos, CPF:00567556867, http://lattes.cnpq.br/9442000157600332, Sales, Maur?cio Pereira de |
| Publisher | Universidade Federal do Rio Grande do Norte, Programa de P?s-Gradua??o em Bioqu?mica, UFRN, BR, Bioqu?mica; Biologia Molecular |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | Unknown |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Format | application/pdf |
| Source | reponame:Repositório Institucional da UFRN, instname:Universidade Federal do Rio Grande do Norte, instacron:UFRN |
| Rights | info:eu-repo/semantics/openAccess |
Page generated in 0.0016 seconds