Mild-acid hydrolysis of a native polysulfated fraction from Acanthophora muscoides generates sulfated oligosaccharides displaying in vitro thrombin generation inhibition
Autor(a) principal: | |
---|---|
Data de Publicação: | 2016 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Acta Scientiarum Biological Sciences |
Texto Completo: | http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/28257 |
Resumo: | Acanthophora muscoides (Rhodophyta) has three polysulfated fractions (Am-1, Am-2 and Am-3). Am-2 displayed anti-inflammation and serpin-independent anticoagulation; however, no effect of oligomers on thrombin-generation (TG) assay has been demonstrated. This study employed mild-acid hydrolysis to obtain low-molecular-size derivatives from Am-2 and compared in vitro inhibitory effects between intact Am-2 and its hydrolysates on a TG assay. The enzymatic extract was fractionated by DEAE-cellulose that revealed Am-2 eluted with 0.75-M NaCl containing sulfate (23%), hexoses (51%) and devoid of proteins, and indicating, by one-dimension nuclear magnetic resonance, structure of galactan similar to that of extract. The depolymerization with HCl (0.02 or 0.04-M, 60°C) for different times progressively reduced the charge density and the molecular-size of Am-2 based on electrophoreses in agarose and polyacrylamide gels, respectively, where at higher acid concentration and critical time up to 5 h yielded fragment of ̴ 10-kDa similar to that of unfractionated heparin (UHEP). Regarding the TG assay, intact Am-2 inhibited concentration-dependent the intrinsic pathway, whereas its hydrolysates abolished it like UHEP, when in 60-fold diluted human plasma using chromogenic method by a continuous system. The results reveal an alternative approach for producing oligosaccharides from A. muscoides with TG inhibition. |
id |
UEM-1_6ecaded5a00dc28e97c43daff9937ba0 |
---|---|
oai_identifier_str |
oai:periodicos.uem.br/ojs:article/28257 |
network_acronym_str |
UEM-1 |
network_name_str |
Acta Scientiarum Biological Sciences |
repository_id_str |
|
spelling |
Mild-acid hydrolysis of a native polysulfated fraction from Acanthophora muscoides generates sulfated oligosaccharides displaying in vitro thrombin generation inhibitionMild-acid hydrolysis of a native polysulfated fraction from Acanthophora muscoides generates sulfated oligosaccharides displaying in vitro thrombin generation inhibitionRhodophyceaepoli-aniônicosdepolimerizaçãotrombina.Química de macromoléculasRhodophyceaepolyanionicsdepolymerizationthrombinAcanthophora muscoides (Rhodophyta) has three polysulfated fractions (Am-1, Am-2 and Am-3). Am-2 displayed anti-inflammation and serpin-independent anticoagulation; however, no effect of oligomers on thrombin-generation (TG) assay has been demonstrated. This study employed mild-acid hydrolysis to obtain low-molecular-size derivatives from Am-2 and compared in vitro inhibitory effects between intact Am-2 and its hydrolysates on a TG assay. The enzymatic extract was fractionated by DEAE-cellulose that revealed Am-2 eluted with 0.75-M NaCl containing sulfate (23%), hexoses (51%) and devoid of proteins, and indicating, by one-dimension nuclear magnetic resonance, structure of galactan similar to that of extract. The depolymerization with HCl (0.02 or 0.04-M, 60°C) for different times progressively reduced the charge density and the molecular-size of Am-2 based on electrophoreses in agarose and polyacrylamide gels, respectively, where at higher acid concentration and critical time up to 5 h yielded fragment of ̴ 10-kDa similar to that of unfractionated heparin (UHEP). Regarding the TG assay, intact Am-2 inhibited concentration-dependent the intrinsic pathway, whereas its hydrolysates abolished it like UHEP, when in 60-fold diluted human plasma using chromogenic method by a continuous system. The results reveal an alternative approach for producing oligosaccharides from A. muscoides with TG inhibition.A. muscoides (Rhodophyta) has three polysulfated fractions (-1, -2 and Am-3). Am-2 displayed anti-inflammation and serpin-independent anticoagulation effects; however, no effect of oligomers on thrombin-generation (TG) has been demonstrated. This study employed mild-acid hydrolysis to obtain low-molecular-size derivatives from Am-2 and compared in vitro inhibitory effects between intact Am-2 and its hydrolysates on a TG assay. The polysaccharidic extract was fractionated by DEAE-cellulose that revealed Am-2 eluted with 0.75-M NaCl containing sulfate (23%), hexoses (51%) and absence of proteins, and indicating, by one-dimension nuclear magnetic resonance, structure of galactan similar to that of the extract. The depolymerization with HCl (0.02 or 0.04-M, 60°C) for different times progressively reduced the charge density and the molecular-size of Am-2 based on electrophoresis in agarose and polyacrylamide gels, respectively, where at higher acid concentration and critical time up to 5h yielded fragment of ̴ 14-kDa similar to that of unfractionated heparin (UHEP). Regarding the TG assay, intact Am-2 inhibited concentration-dependent intrinsic pathway, whereas its hydrolysates abolished it like UHEP, except the analog fragment (92.87% inhibition), when in 60-fold diluted human plasma using chromogenic method in a continuous system. The results reveal an alternative approach for the production of oligosaccharides from A. muscoides with TG inhibition. Universidade Estadual De Maringá2016-07-21info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/2825710.4025/actascibiolsci.v38i1.28257Acta Scientiarum. Biological Sciences; Vol 38 No 1 (2016); 7-15Acta Scientiarum. Biological Sciences; v. 38 n. 1 (2016); 7-151807-863X1679-9283reponame:Acta Scientiarum Biological Sciencesinstname:Universidade Estadual de Maringá (UEM)instacron:UEMenghttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/28257/pdfRodrigues, José Ariévilo GurgelQueiroz, Ismael Nilo Lino deQuinderé, Ana Luíza GomesBenevides, Norma Maria BarrosTovar, Ana Maria FreireMourão, Paulo Antônio de Souzainfo:eu-repo/semantics/openAccess2022-02-17T22:01:02Zoai:periodicos.uem.br/ojs:article/28257Revistahttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSciPUBhttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/oai||actabiol@uem.br1807-863X1679-9283opendoar:2022-02-17T22:01:02Acta Scientiarum Biological Sciences - Universidade Estadual de Maringá (UEM)false |
dc.title.none.fl_str_mv |
Mild-acid hydrolysis of a native polysulfated fraction from Acanthophora muscoides generates sulfated oligosaccharides displaying in vitro thrombin generation inhibition Mild-acid hydrolysis of a native polysulfated fraction from Acanthophora muscoides generates sulfated oligosaccharides displaying in vitro thrombin generation inhibition |
title |
Mild-acid hydrolysis of a native polysulfated fraction from Acanthophora muscoides generates sulfated oligosaccharides displaying in vitro thrombin generation inhibition |
spellingShingle |
Mild-acid hydrolysis of a native polysulfated fraction from Acanthophora muscoides generates sulfated oligosaccharides displaying in vitro thrombin generation inhibition Rodrigues, José Ariévilo Gurgel Rhodophyceae poli-aniônicos depolimerização trombina. Química de macromoléculas Rhodophyceae polyanionics depolymerization thrombin |
title_short |
Mild-acid hydrolysis of a native polysulfated fraction from Acanthophora muscoides generates sulfated oligosaccharides displaying in vitro thrombin generation inhibition |
title_full |
Mild-acid hydrolysis of a native polysulfated fraction from Acanthophora muscoides generates sulfated oligosaccharides displaying in vitro thrombin generation inhibition |
title_fullStr |
Mild-acid hydrolysis of a native polysulfated fraction from Acanthophora muscoides generates sulfated oligosaccharides displaying in vitro thrombin generation inhibition |
title_full_unstemmed |
Mild-acid hydrolysis of a native polysulfated fraction from Acanthophora muscoides generates sulfated oligosaccharides displaying in vitro thrombin generation inhibition |
title_sort |
Mild-acid hydrolysis of a native polysulfated fraction from Acanthophora muscoides generates sulfated oligosaccharides displaying in vitro thrombin generation inhibition |
author |
Rodrigues, José Ariévilo Gurgel |
author_facet |
Rodrigues, José Ariévilo Gurgel Queiroz, Ismael Nilo Lino de Quinderé, Ana Luíza Gomes Benevides, Norma Maria Barros Tovar, Ana Maria Freire Mourão, Paulo Antônio de Souza |
author_role |
author |
author2 |
Queiroz, Ismael Nilo Lino de Quinderé, Ana Luíza Gomes Benevides, Norma Maria Barros Tovar, Ana Maria Freire Mourão, Paulo Antônio de Souza |
author2_role |
author author author author author |
dc.contributor.author.fl_str_mv |
Rodrigues, José Ariévilo Gurgel Queiroz, Ismael Nilo Lino de Quinderé, Ana Luíza Gomes Benevides, Norma Maria Barros Tovar, Ana Maria Freire Mourão, Paulo Antônio de Souza |
dc.subject.por.fl_str_mv |
Rhodophyceae poli-aniônicos depolimerização trombina. Química de macromoléculas Rhodophyceae polyanionics depolymerization thrombin |
topic |
Rhodophyceae poli-aniônicos depolimerização trombina. Química de macromoléculas Rhodophyceae polyanionics depolymerization thrombin |
description |
Acanthophora muscoides (Rhodophyta) has three polysulfated fractions (Am-1, Am-2 and Am-3). Am-2 displayed anti-inflammation and serpin-independent anticoagulation; however, no effect of oligomers on thrombin-generation (TG) assay has been demonstrated. This study employed mild-acid hydrolysis to obtain low-molecular-size derivatives from Am-2 and compared in vitro inhibitory effects between intact Am-2 and its hydrolysates on a TG assay. The enzymatic extract was fractionated by DEAE-cellulose that revealed Am-2 eluted with 0.75-M NaCl containing sulfate (23%), hexoses (51%) and devoid of proteins, and indicating, by one-dimension nuclear magnetic resonance, structure of galactan similar to that of extract. The depolymerization with HCl (0.02 or 0.04-M, 60°C) for different times progressively reduced the charge density and the molecular-size of Am-2 based on electrophoreses in agarose and polyacrylamide gels, respectively, where at higher acid concentration and critical time up to 5 h yielded fragment of ̴ 10-kDa similar to that of unfractionated heparin (UHEP). Regarding the TG assay, intact Am-2 inhibited concentration-dependent the intrinsic pathway, whereas its hydrolysates abolished it like UHEP, when in 60-fold diluted human plasma using chromogenic method by a continuous system. The results reveal an alternative approach for producing oligosaccharides from A. muscoides with TG inhibition. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-07-21 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/28257 10.4025/actascibiolsci.v38i1.28257 |
url |
http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/28257 |
identifier_str_mv |
10.4025/actascibiolsci.v38i1.28257 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/28257/pdf |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Estadual De Maringá |
publisher.none.fl_str_mv |
Universidade Estadual De Maringá |
dc.source.none.fl_str_mv |
Acta Scientiarum. Biological Sciences; Vol 38 No 1 (2016); 7-15 Acta Scientiarum. Biological Sciences; v. 38 n. 1 (2016); 7-15 1807-863X 1679-9283 reponame:Acta Scientiarum Biological Sciences instname:Universidade Estadual de Maringá (UEM) instacron:UEM |
instname_str |
Universidade Estadual de Maringá (UEM) |
instacron_str |
UEM |
institution |
UEM |
reponame_str |
Acta Scientiarum Biological Sciences |
collection |
Acta Scientiarum Biological Sciences |
repository.name.fl_str_mv |
Acta Scientiarum Biological Sciences - Universidade Estadual de Maringá (UEM) |
repository.mail.fl_str_mv |
||actabiol@uem.br |
_version_ |
1799317391009644544 |