Screening, production and biochemical characterization of a new fibrinolytic enzyme produced by Streptomyces sp. (Streptomycetaceae) isolated from Amazonian lichens
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | , , , , , , |
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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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 |
_version_ |
1752129840387981312 |