Assessment Of Nutrients And Study On Polysaccharides From Cereus Sinensis

In recent years anemones are considered to have great medicinal value, and has become dishes on the table. As one research hotspot of marine natural products, sea anemones have a variety of effects. This paper for theory of product development provided an overview of research on biological active substances of sea anemones and polysaccharides, and pointed out the future direction of research and development. International standard and national standard methods were used for determining the nutritional composition, and non-volatile flavor compounds of anemones in order to make the evaluations. Extraction by water and precipitation via alcohol were used to extract anemones polysaccharides. To determine the optimum conditions of polysaccharides extraction, response surface methodology was adopted. Anemones polysaccharides were gradually isolated and purified by DEAE-52 cellulose ion-exchange column and Sephadex G-100 gel column. The immune regulation and cytotoxicity of anemone polysaccharide were also investigated.The objective of this study was to determine the content of chemical compounds(water, ash, lipid, protein, carbohydrate, amino acid, fatty acid and mineral) and assess their nutritive value of Cereus sinensis. The result showed that total protein content was 390 g/kg dry weight(DW), lipid content was 50 g/kg DW, ash content was 167 g/kg DW, carbohydrate content was 260 g/kg DW, fatty acid content was 7.5 g/kg DW, and amino acid content was 277.9 g/kg DW for Cereus sinensis. Cereus sinensis contained variety of amino acid, and meet the amino acid requirements of adults with two exception, isoleucine and valine. Various minerals were contained in Cereus sinensis, such as Ca(3650 mg/kg), Mg(1090 mg/kg) and Se(6.48 mg/kg). Glutamic acid and arginine were the main free amino acids which contributed to pleasant flavors. The taste activity valuation from high to low was GMP, IMP and AMP. Equivalent umami concentration was 1.05 g MSG/100 g. To conclude, Cereus sinensis was a good source of nutritive diet and had high flavor quality.Anemone polysaccharides was extracted by water extraction and alcohol precipitation. The condition of response surface was determined by single factor test, including three main factors as follows: solid-liquid ratio, extraction time and temperature, and three different levels. Box-Behnken software was used to design 17 sets of experiments for determining the optimum extraction conditions of anemone polysaccharides, and binary linear regression equation model was established as a result. The results showed that there was a significant effect on the interaction terms of the extraction rate, including polysaccharide liquid ratio, extraction time and temperature. Finally verified by experiments, the optimum extraction condition of anemone polysaccharide were solid-liquid ratio of 80: 1, bath temperature 72 ℃, extraction time 3 h, and polysaccharide yield was 23.53% under this condition.After removal of protein by TCA method, Cereus sinensis polysaccharides were isolated and purified by chromatography column anemones. Respectively, they flowed through DEAE-52 cellulose anion-exchange chromatography column with the NaCl solution and two components were obtained, CPS-I and CPS-II with different electric charge. And then after concentration, dialysis and freeze-drying process, further purification by sephadex G-100 was performed to obtain pure two Cereus sinensis polysaccharide, CPSI-1 and CPSII-2 with different molecular weight.After the establishment of LPS-induced macrophage RAW264.7 cells model, the Griess reagent was used to assay the yield of NO release, inflammatory cytokines from model cells by the way of CPSI-1 inhibition. And RAW264.7 cells cytotoxicity on CPSI-1 was tested with Alamar Blue reagent. The results showed that compared with the controlled cells, LPS stimulation, CPSI-1 significantly inhibited the amount of NO release from cell model(P <0.01). And the grade level of CPSI-1 cytotoxicity were 0 and 1. Therefore, anemone polysaccharide may exert the bioactivity of immunomodulatory by an inhibition of the production of NO from macrophages models.