Agaricus blazei Murill: isolamento, caracterização estrutural e atividades biológicas dos polissacarídeos e/ou glicoconjugados presentes

Abstract

The use of mushrooms as nutritional and medicinal through the milenar culture in oriental has arrived progressively in the Occident, take by the necessity of the population to search of a more healthy nutrition. Further more, the scientific community has revealed through publications, important findings in relation to the nutritional values and the therapeutic activities of mushrooms. As a contribution for a better knowledge of the properties of this fungus, studies are being tackled with Agaricus blazei Murill cultivated in Brazil in two distinct regions: Piedade, in the state of São Paulo and Guaramiranga, in the state of Ceará. In the first stage, the studies were devoted to the cultivated mushrooms in Ceará (Cog-Ce). The results of the spectroscopy analysis (FTIR and NMR) demonstrated that the purified polysaccharides and fractions (F1 and F4) showed characteristics of the presence of carbohydrates and proteins. After the fractionation was completed, through successive precipitations in ethanol, it was observed by the FTIR spectrum, a reduction of the carbohydrate content and an increase in the protein content. Molecular absorption of the samples indicated the absence of free protein in the purificated polysaccharides and its fractions, which point to the existence of the effective link between carbohydrate and protein, characteristics of the proteoglucans. The analyses revealed the presence of glucans of configuration b(1®6) and a(1®4), with the first having the most representative content. The glucose monosaccharides were predominant between the sugars present and the mineral phosphorus element was the major constituent between the elements analysed. The study of the purified polysaccharides and the isolated fractions of the Agaricus blazei Murill, cultivated in São Paulo (Cog-SP) were carried out in a second stage of the research. The isolated purified polysaccharides, after analyses of the constituents, were characterized and its properties in solutions were investigated, taking into consideration physical-chemical parameters and rheological properties. Furthermore, biological assays were also carried out for evaluation in the case of therapeutic activities. The analyses by 1H and 13C NMR made evident the presence of b glucans (1®6) and a(1®4), which the glucana b(1®6) was predominant. The presence of glucans a(1®4) were greater in relation to the presented content by Cog- Ce. The glucose monosaccharides and the mineral phosphorus element were also predominants in Cog-SP, in relation to the sugars and the minerals present respectively. The greater protein content and the smallest carbohydrate content were found for the purified polysaccharides of Cog-SP when compared to the Cog-Ce. Glucans b(1®6) were isolated through the specific purification procedure with the use of Timol. FTIR and 1H, 13C NMR and the bidimensional COSY and HMQC, confirmed the purity of the material in the b(1®6) configuration, with indication of the link to protein group. The study of purified polysaccharides in solutions showed compounds with high molecular weight. The confirmation of random coils was strongly suggested by light scattering, which was, probably, due to the presence of protein and mineral elements. Low viscosities were observed in the polysaccharides when compared to those of the glucans traditionally used in foods. Rheological measurements demonstrated a Newtonian behavior of the materials. Physical-chemical parameters, such as surface tension and critical micelle concentration (cmc), as well as others, demonstrated similar characteristics to those found for polymers with tensoactive properties. Due to these characteristics, the purified polysaccharides were tested in relation to the solubilizing capacity using the poorly water soluble drug griseofulvin as a model solubilizate. The preliminary test suggest that these polysaccharides, in the gluco-protein form, in solution, are potentially useful to act as a solubilizer for aromatic drugs. The biological test carried out with the purified polysaccharides and/or proteoglucans, demonstrated that they present anti-tumoral activities in vitro, through inhibition of the replication of poliovirus and anti-tumoral in vivo against the Sarcoma 180.