Purificação, caracterização bioquímica e efeito na inibição de biofilme bacteriano de uma lectina presente na esponja marinha chondrilla caribensis

Detalhes bibliográficos
Ano de defesa: 2017
Autor(a) principal: Marques, Dayara Normando
Orientador(a): Sampaio, Alexandre Holanda
Banca de defesa: Não Informado pela instituição
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Não Informado pela instituição
Programa de Pós-Graduação: Não Informado pela instituição
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
Palavras-chave em Português:
Link de acesso: http://www.repositorio.ufc.br/handle/riufc/23668
Resumo: The sponges are included in the phylum Porífera, are sessile, filtering and multicellular animals. They have great biotechnological and pharmacological potential. Several compounds have been discovered in these animals, such as metabolites with anti-inflammatory activity, antimicrobial activity, antitumor activity, antifouling activity, cytotoxic activity and lectin. Lectins can be defined as proteins or glycoproteins that bind reversibly to carbohydrates and are capable of binding erythrocytes and other cell types and / or precipitating polysaccharides. Marine sponges are considered a rich source of lectins, with great biotechnological potential. Thus, the objective of the present work was to purify, characterize biochemically and evaluate the effect on bacterial biofilm inhibition of an isolated lectin in the marine sponge Chodrilla caribensis from the coast of Ceará. CCL (Chodrilla caribensis lectin) was purified from the combination of ion exchange chromatography and affinity chromatography. The new lectin is a glycoprotein, with a content of 5.2% carbohydrate and a size of approximately 15 kDa. CCL showed affinity for lactose, its activity was better at pH in the neutral-alkaline range and the protein only lost its activity completely when heated to 100 ° C. The theoretical secondary structure of CCL consisted of 10.4% of α-helix, 73.4% of β-structure and 16.2% of unstructured region. In addition, the CCL was crystallized through the preliminary crystallization test. The protein was highly toxic against Artemia sp. Nauplii, presenting an LC 50 = 6.3 μg.mL-1 and was effective in inhibiting the biofilm formation of Gram-positive bacteria, S. aureus and S. epidermidis. Regarding the number of viable cells, CCL was effective only in treatment with S. epidermidis, at all concentrations.
id UFC-7_9a3e38f7d23b886b5cb7c4bb5d4efd96
oai_identifier_str oai:repositorio.ufc.br:riufc/23668
network_acronym_str UFC-7
network_name_str Repositório Institucional da Universidade Federal do Ceará (UFC)
repository_id_str
spelling Marques, Dayara NormandoSampaio, Alexandre Holanda2017-06-27T22:16:32Z2017-06-27T22:16:32Z2017MARQUES, Dayara Normando Marques. Purificação, caracterização bioquímica e efeito na inibição de biofilme bacteriano de uma lectina presente na esponja marinha chondrilla caribensis. 2017. 54 f. Dissertação (Mestrado em Engenharia de Pesca)- Universidade Federal do Ceará, Fortaleza, 2017.http://www.repositorio.ufc.br/handle/riufc/23668The sponges are included in the phylum Porífera, are sessile, filtering and multicellular animals. They have great biotechnological and pharmacological potential. Several compounds have been discovered in these animals, such as metabolites with anti-inflammatory activity, antimicrobial activity, antitumor activity, antifouling activity, cytotoxic activity and lectin. Lectins can be defined as proteins or glycoproteins that bind reversibly to carbohydrates and are capable of binding erythrocytes and other cell types and / or precipitating polysaccharides. Marine sponges are considered a rich source of lectins, with great biotechnological potential. Thus, the objective of the present work was to purify, characterize biochemically and evaluate the effect on bacterial biofilm inhibition of an isolated lectin in the marine sponge Chodrilla caribensis from the coast of Ceará. CCL (Chodrilla caribensis lectin) was purified from the combination of ion exchange chromatography and affinity chromatography. The new lectin is a glycoprotein, with a content of 5.2% carbohydrate and a size of approximately 15 kDa. CCL showed affinity for lactose, its activity was better at pH in the neutral-alkaline range and the protein only lost its activity completely when heated to 100 ° C. The theoretical secondary structure of CCL consisted of 10.4% of α-helix, 73.4% of β-structure and 16.2% of unstructured region. In addition, the CCL was crystallized through the preliminary crystallization test. The protein was highly toxic against Artemia sp. Nauplii, presenting an LC 50 = 6.3 μg.mL-1 and was effective in inhibiting the biofilm formation of Gram-positive bacteria, S. aureus and S. epidermidis. Regarding the number of viable cells, CCL was effective only in treatment with S. epidermidis, at all concentrations.As esponjas estão incluídas no filo Porífera, são animais sésseis, filtradores e multicelulares. Elas possuem um grande potencial biotecnológico e farmacológico. Vários compostos têm sido descobertos nesses animais, tais como metabolitos com atividade anti-inflamatória, atividade antimicrobiana, atividade antitumoral, atividade anti-incrustante, atividade citotóxica e lecinas. As lectinas podem ser definidas como proteínas ou glicoproteínas que se ligam reversivelmente aos carboidratos e são capazes de aglutinar eritrócitos e outros tipos de células e/ou precipitar polissacarídeos. As esponjas marinhas são consideradas uma rica fonte de lectinas, com grande potencial biotecnológico. Dessa forma, objetivo do presente trabalho foi purificar, caracterizar bioquimicamente e avaliar o efeito na inibição do biofilme bacteriano de uma lectina isolada na esponja marinha Chodrilla caribensis do litoral do Ceará. CCL (Chodrilla caribensis lectin) foi purificada a partir da combinação de cromatografia de troca iônica e cromatografia de afinidade. A nova lectina é uma glicoproteina, com um teor de 5,2% de carboidratos e com um tamanho de aproximadamente 15 kDa. CCL apresentou afinidade por lactose, sua atividade foi melhor em pH na faixa neutro-alcalina e a perdeu sua atividade completamente quando aquecida a 100°C. A estrutura teórica secundária de CCL consistiu em 10,4% de α hélice, 73,4% de estrutura β e 16,2 % de região desestruturada. Além disso, a CCL foi cristalizada, através do ensaio preliminar de cristalização. A proteína foi altamente tóxica contra naúplios de Artemia sp., apresentando um LC50 = 6,3 μg.mL-1 e foi eficaz na inibição da formação de biofilme das bactérias Gram-positivas, S. aureus e S. epidermidis. Em relação ao número de células viáveis, CCL foi eficaz no tratamento com S. epidermidis, em todas as concentrações.LectinaEsponja marinhaBiofilmePurificação, caracterização bioquímica e efeito na inibição de biofilme bacteriano de uma lectina presente na esponja marinha chondrilla caribensisPurification, biochemical characterization and effect on the inhibition of bacterial biofilm of a lectin present in the marine sponge chondrilla caribensisinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisporreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccessORIGINAL2017_dis_dnmarques.pdf2017_dis_dnmarques.pdfapplication/pdf1163340http://repositorio.ufc.br/bitstream/riufc/23668/1/2017_dis_dnmarques.pdf0e0999c264fccdb1643cbc1d0a0fca68MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.ufc.br/bitstream/riufc/23668/2/license.txt8a4605be74aa9ea9d79846c1fba20a33MD52riufc/236682020-05-14 15:45:09.258oai:repositorio.ufc.br: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Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2020-05-14T18:45:09Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.pt_BR.fl_str_mv Purificação, caracterização bioquímica e efeito na inibição de biofilme bacteriano de uma lectina presente na esponja marinha chondrilla caribensis
dc.title.en.pt_BR.fl_str_mv Purification, biochemical characterization and effect on the inhibition of bacterial biofilm of a lectin present in the marine sponge chondrilla caribensis
title Purificação, caracterização bioquímica e efeito na inibição de biofilme bacteriano de uma lectina presente na esponja marinha chondrilla caribensis
spellingShingle Purificação, caracterização bioquímica e efeito na inibição de biofilme bacteriano de uma lectina presente na esponja marinha chondrilla caribensis
Marques, Dayara Normando
Lectina
Esponja marinha
Biofilme
title_short Purificação, caracterização bioquímica e efeito na inibição de biofilme bacteriano de uma lectina presente na esponja marinha chondrilla caribensis
title_full Purificação, caracterização bioquímica e efeito na inibição de biofilme bacteriano de uma lectina presente na esponja marinha chondrilla caribensis
title_fullStr Purificação, caracterização bioquímica e efeito na inibição de biofilme bacteriano de uma lectina presente na esponja marinha chondrilla caribensis
title_full_unstemmed Purificação, caracterização bioquímica e efeito na inibição de biofilme bacteriano de uma lectina presente na esponja marinha chondrilla caribensis
title_sort Purificação, caracterização bioquímica e efeito na inibição de biofilme bacteriano de uma lectina presente na esponja marinha chondrilla caribensis
author Marques, Dayara Normando
author_facet Marques, Dayara Normando
author_role author
dc.contributor.author.fl_str_mv Marques, Dayara Normando
dc.contributor.advisor1.fl_str_mv Sampaio, Alexandre Holanda
contributor_str_mv Sampaio, Alexandre Holanda
dc.subject.por.fl_str_mv Lectina
Esponja marinha
Biofilme
topic Lectina
Esponja marinha
Biofilme
description The sponges are included in the phylum Porífera, are sessile, filtering and multicellular animals. They have great biotechnological and pharmacological potential. Several compounds have been discovered in these animals, such as metabolites with anti-inflammatory activity, antimicrobial activity, antitumor activity, antifouling activity, cytotoxic activity and lectin. Lectins can be defined as proteins or glycoproteins that bind reversibly to carbohydrates and are capable of binding erythrocytes and other cell types and / or precipitating polysaccharides. Marine sponges are considered a rich source of lectins, with great biotechnological potential. Thus, the objective of the present work was to purify, characterize biochemically and evaluate the effect on bacterial biofilm inhibition of an isolated lectin in the marine sponge Chodrilla caribensis from the coast of Ceará. CCL (Chodrilla caribensis lectin) was purified from the combination of ion exchange chromatography and affinity chromatography. The new lectin is a glycoprotein, with a content of 5.2% carbohydrate and a size of approximately 15 kDa. CCL showed affinity for lactose, its activity was better at pH in the neutral-alkaline range and the protein only lost its activity completely when heated to 100 ° C. The theoretical secondary structure of CCL consisted of 10.4% of α-helix, 73.4% of β-structure and 16.2% of unstructured region. In addition, the CCL was crystallized through the preliminary crystallization test. The protein was highly toxic against Artemia sp. Nauplii, presenting an LC 50 = 6.3 μg.mL-1 and was effective in inhibiting the biofilm formation of Gram-positive bacteria, S. aureus and S. epidermidis. Regarding the number of viable cells, CCL was effective only in treatment with S. epidermidis, at all concentrations.
publishDate 2017
dc.date.accessioned.fl_str_mv 2017-06-27T22:16:32Z
dc.date.available.fl_str_mv 2017-06-27T22:16:32Z
dc.date.issued.fl_str_mv 2017
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.citation.fl_str_mv MARQUES, Dayara Normando Marques. Purificação, caracterização bioquímica e efeito na inibição de biofilme bacteriano de uma lectina presente na esponja marinha chondrilla caribensis. 2017. 54 f. Dissertação (Mestrado em Engenharia de Pesca)- Universidade Federal do Ceará, Fortaleza, 2017.
dc.identifier.uri.fl_str_mv http://www.repositorio.ufc.br/handle/riufc/23668
identifier_str_mv MARQUES, Dayara Normando Marques. Purificação, caracterização bioquímica e efeito na inibição de biofilme bacteriano de uma lectina presente na esponja marinha chondrilla caribensis. 2017. 54 f. Dissertação (Mestrado em Engenharia de Pesca)- Universidade Federal do Ceará, Fortaleza, 2017.
url http://www.repositorio.ufc.br/handle/riufc/23668
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.source.none.fl_str_mv reponame:Repositório Institucional da Universidade Federal do Ceará (UFC)
instname:Universidade Federal do Ceará (UFC)
instacron:UFC
instname_str Universidade Federal do Ceará (UFC)
instacron_str UFC
institution UFC
reponame_str Repositório Institucional da Universidade Federal do Ceará (UFC)
collection Repositório Institucional da Universidade Federal do Ceará (UFC)
bitstream.url.fl_str_mv http://repositorio.ufc.br/bitstream/riufc/23668/1/2017_dis_dnmarques.pdf
http://repositorio.ufc.br/bitstream/riufc/23668/2/license.txt
bitstream.checksum.fl_str_mv 0e0999c264fccdb1643cbc1d0a0fca68
8a4605be74aa9ea9d79846c1fba20a33
bitstream.checksumAlgorithm.fl_str_mv MD5
MD5
repository.name.fl_str_mv Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)
repository.mail.fl_str_mv bu@ufc.br || repositorio@ufc.br
_version_ 1847793340714582016