<b>Structural analysis of a sulfated polysaccharidic fraction obtained from the coenocytic green seaweed <i>Caulerpa cupressoides<i> var. <i>lycopodium<i> </b>

Abstract

ABSTRACT.Researches on structural chemistry of sulfated polysaccharides (SPs) have been mainly focused on red and brown algae. Caulerpa cupressoides var. lycopodium (Chlorophyta) contains three SPs fractions (Cc-SP 1 , Cc-SP 2 and Cc-SP 3 ). Cc-SP 1 and Cc-SP 2 had anticoagulant (in vitro) and anti-and prothrombotic, antinociceptive and/or anti-inflammatory (in vivo) effects. However, their structural features have not yet been investigated. This study analyzed the chemical composition, elemental microanalysis and structural features by infrared (IR) and nuclear magnetic resonance ( 1 H NMR) spectroscopy of Cc-SPs. Fractionation of SPs by DEAE-cellulose yielded Cc-SP 1 , Cc-SP 2 and CcSP 3 containing differences among the relative proportions of sulfate (14.67-26.72%), total sugars (34.92-49.73%) and uronic acid (7.15-7.22%). Carbon (21.76-29.62%), sulfate (2.16-4.55%), nitrogen (0.85-1.57%) and hydrogen (4.57-5.86%) contents were obtained using a CHN equipment. Data from IR indicated occurrence of sulfate ester, galactose-6-sulfate, uronic acid and glycoside linkages. For 1 H NMR spectrum of the soluble Cc-SP1 fraction, it was mainly found β-galactopyranose residues and CH 3 group. The results showed that Cc-SPs fractions have some structural features similar to others studied Caulerpaceae SPs.Keywords: Chlorophyta, Caulerpaceae, sulfated polymers, chemical analysis, structural chemistry.Análise estrutural de uma fração polissacarídica sulfatada obtida da alga marinha verde cenocítica Caulerpa cupressoides var. lycopodium RESUMO. Pesquisas sobre química estrutural de polissacarídeos sulfatados (PSs) têm focado principalmente em algas vermelhas e pardas. A Chlorophyta Caulerpa cupressoides var. lycopodium contém três frações de PSs (Cc-PS 1 ; Cc-PS 2 e Cc-PS 3 ). As Cc-PS 1 e Cc-PS 2 apresentaram efeitos anticoagulante (in vitro), anti-e pró-trombótico, antinociceptivo e/ou anti-inflamatório (in vivo). Entretanto, ainda não foram investigadas suas características estruturais. Este estudo analisou de Cc-PSs a composição química, microanálise elementar e características estruturais por infravermelho (IV) e espectroscopia de ressonância magnética nuclear (RMN 1 H). O fracionamento dos PSs por DEAE-celulose rendeu Cc-PS 1 ; Cc-PS 2 e Cc-PS 3 contendo diferenças entre as proporções relativas de sulfato (14,67-26,72%), açúcares totais (34,92-49,73%) e ácido urônico (7,15-7,22%). Foram obtidos, usando um equipamento CHN, conteúdos de carbono (7,15-7,22%), sulfato (2,16-4,55%), nitrogênio (7,15-7,22%) e hidrogênio (7,15-7,22%). O IV forneceu dados sobre a ocorrência de éster sulfato, galactose-6-sulfato, ácido urônico e ligações glicosídicas. Foram encontrados principalmente para o espectro de RMN 1 H da fração solúvel Cc-PS 1 , resíduos de β-galactopiranoses e de grupo CH 3 . Os resultados mostraram frações de Cc-PSs detentoras de algumas características estruturais semelhantes aos de outros PSs de Caulerpaceae estudados.Palavras-chave: Chlorophyta, Caulerpaceae, polímeros sulfatados, análise química, química estrutural.

<b>Extraction and structural properties of <i>Acanthophora muscoides </i>(Rhodophyceae) extracellular matrix sulfated polysaccharides and their effects on coagulation

“…Spectra also presented two singlets at δ H 3.7 ppm related to 2-O-methyl groups (Fenoradosoa et al, 2009) and another signal at δ H 1.7 ppm indicative of CH 3 groups (Rodrigues et al, 2014), which would correspond, respectively, the chemical shifts of a methyl-esterified C 2 position, as well as from the CH 3 groups themselves, as confirmed in another investigation (Quinderé et al, 2014). An attempt to assign the occurrence of pyruvic acid in polysaccharidic extracts composition was further suggested with basis on variable intensity resonance signals around δ H 1.7 ppm (Fenoradosoa et al, 2009).…”

“…However, very complex spectra showing several unresolved peaks were recorded and the 1 H-signals for each native extract were displayed from δ H 5.6 to 1.6 ppm ( Figures 1A, B and C), as based on Quinderé et al (2014) who previously analyzed a native fraction from this same algal species with high degree of structural heterogeneity, typical for algae SPs (Maciel et al, 2008;Fenoradosoa et al, 2009;Rodrigues et al, 2014).…”

“…Herein, values would reflex a disorganization of the algal tissue because proteolysis attack the extracellular matrix and cleave the interaction between SPs and proteins (Rodrigues et al, 2011;Fidelis et al, 2014), especially on the surface texture from mucilage deposition on the cell-wall, as observed on vegetative cells in red algae, in which the SPs could be mainly located (Estevez et al, 2008;Miguel et al, 2014); thus, leading to obtain novel biologically active SPs (Rodrigues et al, 2009). Papain-enzyme action, as demonstrated by the Bradford' method, also removed contaminant proteins in all analyzed raw samples (Rodrigues et al, 2011;Araújo et al, 2012;Quinderé et al, 2013;Rodrigues et al, 2014).…”

“…Anticoagulation (Rodrigues et al, 2009;Rodrigues et al, 2013;Mourão, 2015), antinociception and anti-inflammation Rodrigues et al, 2012;Wu et al, 2016) are some of their most important pharmacological properties. Although also occurring in other living organisms (Aquino, Landeira-Fernandez, Valente, Andrade, & Mourão, 2005;Dantas-Santos et al, 2012;Chang, Lur, Lu, & Cheng, 2013), sulfated galactans (mainly carrageenan and agar) (Rhodophyceae) (Estevez et al, 2008;Mourão, 2015), fucan or fuicodan (Phaeophyceae) (Leite et al, 1998;Pereira & Costa-Lotufo, 2012) and other structurally diverse SPs found in Chlorophyceae Rodrigues et al, 2014) are the main classes of these biomolecules present in nature. Yield and physical-chemical and biological properties of SPs depend 1) on the morphology of the thallus; 2) the tissue under study (Estevez et al, 2008;Miguel et al, 2014); 3) the use of different protocols and algal species for extraction (Rodrigues et al, 2009;Rodrigues et al, 2011;Araújo et al, 2012); 4) climatic factors; and 5) growth and/or the reproducible cycle of the algae (Zvyagintseva et al, 2003;Cardozo et al, 2007;Pereira & Costa-Lotufo, 2012).…”

Molecular characteristics of polysulfated fractions from the green seaweed <i>Caulerpa cupressoides</i> and actions on thrombin generation <i>in vitro

“…lycopodium (Vahl) C. Agardh is a particularly interesting benthic species of green seaweed endemic to the Ceará coast, Brazil, having nutritional value (Carneiro, Rodrigues, Teles, Cavalcante, & Benevides, 2014) and metabolites-derived physiological effects (Vanderlei et al, 2010;Rivanor et al, 2014;Wang et al, 2014). Studies performed by Rodrigues et al (2011a), Rodrigues, Quinderé, Queiroz, Coura, and Benevides (2012a), Rodrigues et al (2014a) isolated and partially characterized three SPs fractions (-SP1, -SP2 and Cc-SP3), of which only Cc-SP2 showed sulfation-dependent anticoagulant effect vs. UHEP. Cc-SP1 and Cc-SP3 had no in vitro anti-clotting effects (Rodrigues et al, 2011a), but Cc-SP1 revealed to be an in vitro anti-herpetic and anti-dengue agent (Rodrigues et al, 2017a).…”