| Registro completo |
| Provedor de dados: |
Biological Sciences
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| País: |
Brazil
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| Título: |
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
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| Autores: |
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
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| Data: |
2016-07-21
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| Ano: |
2016
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| Palavras-chave: |
Ciências Biológicas e Ambientais Rhodophyceae
Poli-aniônicos
Depolimerização
Trombina. Química de macromoléculas Rhodophyceae
Polyanionics
Depolymerization
Thrombin
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| 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.
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.
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| Tipo: |
Info:eu-repo/semantics/article
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| Idioma: |
Inglês
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| Identificador: |
http://periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/28257
10.4025/actascibiolsci.v38i1.28257
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| Editor: |
Universidade Estadual De Maringá
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| Relação: |
http://periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/28257/pdf
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| Formato: |
application/pdf
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| Fonte: |
Acta Scientiarum. Biological Sciences; v. 38, n. 1 (2016); 7-15
Acta Scientiarum. Biological Sciences; v. 38, n. 1 (2016); 7-15
1807-863X
1679-9283
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