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Previous issue date: 2015-06-26 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / Sementes de plantas s?o reservat?rios de mol?culas com grande potencial
para elabora??o de bioprodutos e por este motivo uma especial aten??o tem sido
direcionada na busca de prote?nas bioativas com a??o antimetab?lica e propriedades
farmacol?gicas. As lectinas de plantas s?o prote?nas com atividades biol?gicas
relacionadas a defesa, mas que podem ser utilizadas, heterologamente, como
interferentes no metabolismo de outros organismos. Uma lectina das sementes de
Canavalia maritima (CML) foi purificada em dois passos, pelo fracionamento com
sulfato de am?nio seguido pela cromatografia de afinidade em coluna Sephadex G-
50. CML foi analisada por SDS-PAGE e parte de sua sequ?ncia de amino?cidos
determinada por espectrometria de massas. Os 20 res?duos identificados
apresentaram 100% de identidade com ConM. ConM foi testada contra eritr?citos do
sangue perif?rico humano e foi constatado a aus?ncia de toxicidade quando
incubados com at? 1000 ?g/mL da prote?na. J? contra c?lulas mononucleares, a
lectina n?o foi significativamente t?xica nas concentra??es de 1, 2,5 e 5 ?g/mL, mas
o foi a partir da concentra??o 7 ?g/mL e contra a linhagem RAEC a prote?na n?o
apresentou toxicidade significante com 25 ?g/ml da prote?na. Outros ensaios de
citotoxicidade revelaram que a ConM ? t?xica para as linhagens tumorais A549, 786-
0, HT-29 e HeLa no intervalo entre 24 e 72 horas, principalmente nas concentra??es
de 5, 7 e 10 ?g/ml. ConM foi capaz de aglutinar as formas promastigotas de
Leishmania spp na concentra??o de 6,25 ?g/ml. Quanto a a??o contra Candida spp,
nenhuma das concentra??es testadas (1 a 500 ?g/ml) foi capaz de interferir no
crescimento. Quando avaliada a atividade bacteriost?tica, a ConM nas concentra??es
variando de 0,39 a 400 ?g/ml n?o inibiu o crescimento de Escherichia coli, j? para
Staphylococcus aureus a concentra??o de 25 ?g/ml se mostrou ativa, reduzindo o
crescimento em 75 %. Com base nos dados obtidos, a lectina isolada corresponde a
uma isolectina do tipo ConA, e suas propriedades anti-tumoral, bacteriost?tico e
aglutinante para Leishmania precisam ser melhor investigados para que seu potencial
biotecnol?gico seja explorado. / Plant seeds are reservoirs of molecules with great potential for development of
bioproducts and for this reason special attention has been directed in the search of
bioactive proteins with antimetabolic and pharmacological properties action. Plant
lectins are proteins with biological activities related to defense, but that can be used,
heterologously as interfering with the metabolism of other organisms. A Canavalia
mar?tima seeds lectin (CML) was purified in two steps by ammonium sulfate
fractionation followed by affinity chromatography on Sephadex G-50 column. CML was
analyzed by SDS-PAGE and part of its amino acid sequence determined by mass
spectrometry. The 20 resulting residues identified showed 100% identity with conm.
ConM was tested against erythrocytes of human peripheral blood and it was observed
absence of toxicity when incubated with up to 1000 ?g/mL of protein. Since against
mononuclear cells, the lectin was not significantly toxic at concentrations of 1, 2.5 and
5 ?g/mL, but it was from concentration 7 ?g/mL. Against RAEC lineage protein showed
no significant toxicity with 25 ?g/mL of protein. Other cytotoxicity assays revealed that
CONM is toxic for the tumor cell lines A549, 786-0, HT-29 and HeLa in the range
between 24 and 72 hours, particularly at concentrations of 5, 7 and 10 ?g/mL. ConM
was still able to agglutinate promastigotes of Leishmania spp at concentrations of 6.25
?g/mL. About action against Candida spp any of the tested concentrations (1 to 500
?g/ml) was able to interfere with growth. When evaluated bacteriostatic activity, ConM
in concentrations ranging from 0.39 to 400 ?g/mL did not inhibit the growth of
Escherichia coli, but the concentration of 25 ?g/mL proved active aganist
Staphylococcus aureus, reducing its growth by 75%. Based on these data, the lectin
corresponds to a isolectin ConA-like, and its anti-tumor properties, bacteriostatic and
binder for Leishmania need to be further investigated for its biotechnological potential
exploited.
| Identifer | oai:union.ndltd.org:IBICT/oai:repositorio.ufrn.br:123456789/20329 |
| Date | 26 June 2015 |
| Creators | Ara?jo, Jonalson Nogueira |
| Contributors | 58024867320, http://lattes.cnpq.br/6644671747055211, Lima, Jailma Almeida de, 05116158411, http://lattes.cnpq.br/9077722100091492, Lima, Jo?o Paulo Matos Santos, 79332021368, http://lattes.cnpq.br/3289758851760692, Santos, Elizeu Antunes dos, Uch?a, Adriana Ferreira |
| Publisher | Universidade Federal do Rio Grande do Norte, PROGRAMA DE P?S-GRADUA??O EM BIOQU?MICA, UFRN, Brasil |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Source | reponame:Repositório Institucional da UFRN, instname:Universidade Federal do Rio Grande do Norte, instacron:UFRN |
| Rights | info:eu-repo/semantics/openAccess |
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