Screening, production and biochemical characterization of a new fibrinolytic enzyme produced by Streptomyces sp. (Streptomycetaceae) isolated from Amazonian lichens

Detalhes bibliográficos
Autor(a) principal: SILVA,Germana Michelle de Medeiros e
Data de Publicação: 2016
Outros Autores: BEZERRA,Raquel Pedrosa, TEIXEIRA,José António, SILVA,Flávio Oliveira, CORREIA,Juliana Mendes, PORTO,Tatiana Souza, LIMA-FILHO,José Luis, PORTO,Ana Lúcia Figueiredo
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Acta Amazonica
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0044-59672016000300323
Resumo: ABSTRACT Thrombosis is a pathophysiological disorder caused by accumulation of fibrin in the blood. Fibrinolytic proteases with potent thrombolytic activity have been produced by diverse microbial sources. Considering the microbial biodiversity of the Amazon region, this study aimed at the screening, production and biochemical characterization of a fibrinolytic enzyme produced by Streptomyces sp. isolated from Amazonian lichens. The strain Streptomyces DPUA1576 showed the highest fibrinolytic activity, which was 283 mm2. Three variables at two levels were used to assess their effects on the fibrinolytic production. The parameters studied were agitation (0.28 - 1.12 g), temperature (28 - 36 ºC) and pH (6.0 - 8.0); all of them had significant effects on the fibrinolytic production. The maximum fibrinolytic activity (304 mm2) was observed at 1.12 g, 28 ºC, and pH of 8.0. The crude extract of the fermentation broth was used to assess the biochemical properties of the enzyme. Protease and fibrinolytic activities were stable during 6 h, at a pH ranging from 6.8 to 8.4 and 5.8 to 9.2, respectively. Optimum temperature for protease activity ranged between 35 and 55 °C, while the highest fibrinolytic activity was observed at 45 ºC. Proteolytic activity was inhibited by Cu2+ and Co2+ ions, phenylmethylsulfonyl fluoride (PMSF) and pepstatin A, which suggests that the enzyme is a serine protease. Enzymatic extract cleaved fibrinogen at the subunits Aα-chain, Aβ-chain, and γ-chain. The results indicated that Streptomyces sp. DPUA 1576 produces enzymes with fibrinolytic and fibrinogenolytic activity, enzymes with an important application in the pharmaceutical industry.
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spelling Screening, production and biochemical characterization of a new fibrinolytic enzyme produced by Streptomyces sp. (Streptomycetaceae) isolated from Amazonian lichensActinomycetesfibrinolytic proteaseinhibitor proteasefibrinogenolytic activityABSTRACT Thrombosis is a pathophysiological disorder caused by accumulation of fibrin in the blood. Fibrinolytic proteases with potent thrombolytic activity have been produced by diverse microbial sources. Considering the microbial biodiversity of the Amazon region, this study aimed at the screening, production and biochemical characterization of a fibrinolytic enzyme produced by Streptomyces sp. isolated from Amazonian lichens. The strain Streptomyces DPUA1576 showed the highest fibrinolytic activity, which was 283 mm2. Three variables at two levels were used to assess their effects on the fibrinolytic production. The parameters studied were agitation (0.28 - 1.12 g), temperature (28 - 36 ºC) and pH (6.0 - 8.0); all of them had significant effects on the fibrinolytic production. The maximum fibrinolytic activity (304 mm2) was observed at 1.12 g, 28 ºC, and pH of 8.0. The crude extract of the fermentation broth was used to assess the biochemical properties of the enzyme. Protease and fibrinolytic activities were stable during 6 h, at a pH ranging from 6.8 to 8.4 and 5.8 to 9.2, respectively. Optimum temperature for protease activity ranged between 35 and 55 °C, while the highest fibrinolytic activity was observed at 45 ºC. Proteolytic activity was inhibited by Cu2+ and Co2+ ions, phenylmethylsulfonyl fluoride (PMSF) and pepstatin A, which suggests that the enzyme is a serine protease. Enzymatic extract cleaved fibrinogen at the subunits Aα-chain, Aβ-chain, and γ-chain. The results indicated that Streptomyces sp. DPUA 1576 produces enzymes with fibrinolytic and fibrinogenolytic activity, enzymes with an important application in the pharmaceutical industry.Instituto Nacional de Pesquisas da Amazônia2016-09-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0044-59672016000300323Acta Amazonica v.46 n.3 2016reponame:Acta Amazonicainstname:Instituto Nacional de Pesquisas da Amazônia (INPA)instacron:INPA10.1590/1809-4392201600022info:eu-repo/semantics/openAccessSILVA,Germana Michelle de Medeiros eBEZERRA,Raquel PedrosaTEIXEIRA,José AntónioSILVA,Flávio OliveiraCORREIA,Juliana MendesPORTO,Tatiana SouzaLIMA-FILHO,José LuisPORTO,Ana Lúcia Figueiredoeng2016-06-29T00:00:00Zoai:scielo:S0044-59672016000300323Revistahttps://acta.inpa.gov.br/PUBhttps://old.scielo.br/oai/scielo-oai.phpacta@inpa.gov.br||acta@inpa.gov.br1809-43920044-5967opendoar:2016-06-29T00:00Acta Amazonica - Instituto Nacional de Pesquisas da Amazônia (INPA)false
dc.title.none.fl_str_mv Screening, production and biochemical characterization of a new fibrinolytic enzyme produced by Streptomyces sp. (Streptomycetaceae) isolated from Amazonian lichens
title Screening, production and biochemical characterization of a new fibrinolytic enzyme produced by Streptomyces sp. (Streptomycetaceae) isolated from Amazonian lichens
spellingShingle Screening, production and biochemical characterization of a new fibrinolytic enzyme produced by Streptomyces sp. (Streptomycetaceae) isolated from Amazonian lichens
SILVA,Germana Michelle de Medeiros e
Actinomycetes
fibrinolytic protease
inhibitor protease
fibrinogenolytic activity
title_short Screening, production and biochemical characterization of a new fibrinolytic enzyme produced by Streptomyces sp. (Streptomycetaceae) isolated from Amazonian lichens
title_full Screening, production and biochemical characterization of a new fibrinolytic enzyme produced by Streptomyces sp. (Streptomycetaceae) isolated from Amazonian lichens
title_fullStr Screening, production and biochemical characterization of a new fibrinolytic enzyme produced by Streptomyces sp. (Streptomycetaceae) isolated from Amazonian lichens
title_full_unstemmed Screening, production and biochemical characterization of a new fibrinolytic enzyme produced by Streptomyces sp. (Streptomycetaceae) isolated from Amazonian lichens
title_sort Screening, production and biochemical characterization of a new fibrinolytic enzyme produced by Streptomyces sp. (Streptomycetaceae) isolated from Amazonian lichens
author SILVA,Germana Michelle de Medeiros e
author_facet SILVA,Germana Michelle de Medeiros e
BEZERRA,Raquel Pedrosa
TEIXEIRA,José António
SILVA,Flávio Oliveira
CORREIA,Juliana Mendes
PORTO,Tatiana Souza
LIMA-FILHO,José Luis
PORTO,Ana Lúcia Figueiredo
author_role author
author2 BEZERRA,Raquel Pedrosa
TEIXEIRA,José António
SILVA,Flávio Oliveira
CORREIA,Juliana Mendes
PORTO,Tatiana Souza
LIMA-FILHO,José Luis
PORTO,Ana Lúcia Figueiredo
author2_role author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv SILVA,Germana Michelle de Medeiros e
BEZERRA,Raquel Pedrosa
TEIXEIRA,José António
SILVA,Flávio Oliveira
CORREIA,Juliana Mendes
PORTO,Tatiana Souza
LIMA-FILHO,José Luis
PORTO,Ana Lúcia Figueiredo
dc.subject.por.fl_str_mv Actinomycetes
fibrinolytic protease
inhibitor protease
fibrinogenolytic activity
topic Actinomycetes
fibrinolytic protease
inhibitor protease
fibrinogenolytic activity
description ABSTRACT Thrombosis is a pathophysiological disorder caused by accumulation of fibrin in the blood. Fibrinolytic proteases with potent thrombolytic activity have been produced by diverse microbial sources. Considering the microbial biodiversity of the Amazon region, this study aimed at the screening, production and biochemical characterization of a fibrinolytic enzyme produced by Streptomyces sp. isolated from Amazonian lichens. The strain Streptomyces DPUA1576 showed the highest fibrinolytic activity, which was 283 mm2. Three variables at two levels were used to assess their effects on the fibrinolytic production. The parameters studied were agitation (0.28 - 1.12 g), temperature (28 - 36 ºC) and pH (6.0 - 8.0); all of them had significant effects on the fibrinolytic production. The maximum fibrinolytic activity (304 mm2) was observed at 1.12 g, 28 ºC, and pH of 8.0. The crude extract of the fermentation broth was used to assess the biochemical properties of the enzyme. Protease and fibrinolytic activities were stable during 6 h, at a pH ranging from 6.8 to 8.4 and 5.8 to 9.2, respectively. Optimum temperature for protease activity ranged between 35 and 55 °C, while the highest fibrinolytic activity was observed at 45 ºC. Proteolytic activity was inhibited by Cu2+ and Co2+ ions, phenylmethylsulfonyl fluoride (PMSF) and pepstatin A, which suggests that the enzyme is a serine protease. Enzymatic extract cleaved fibrinogen at the subunits Aα-chain, Aβ-chain, and γ-chain. The results indicated that Streptomyces sp. DPUA 1576 produces enzymes with fibrinolytic and fibrinogenolytic activity, enzymes with an important application in the pharmaceutical industry.
publishDate 2016
dc.date.none.fl_str_mv 2016-09-01
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0044-59672016000300323
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0044-59672016000300323
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/1809-4392201600022
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv text/html
dc.publisher.none.fl_str_mv Instituto Nacional de Pesquisas da Amazônia
publisher.none.fl_str_mv Instituto Nacional de Pesquisas da Amazônia
dc.source.none.fl_str_mv Acta Amazonica v.46 n.3 2016
reponame:Acta Amazonica
instname:Instituto Nacional de Pesquisas da Amazônia (INPA)
instacron:INPA
instname_str Instituto Nacional de Pesquisas da Amazônia (INPA)
instacron_str INPA
institution INPA
reponame_str Acta Amazonica
collection Acta Amazonica
repository.name.fl_str_mv Acta Amazonica - Instituto Nacional de Pesquisas da Amazônia (INPA)
repository.mail.fl_str_mv acta@inpa.gov.br||acta@inpa.gov.br
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