| Mosla chinensis Maxim. cv. jiangxiangru is mainly distributed in Jiangxiprovince. Also as the unique resource in Jiangxi, it belongs to the member of Labiataefamily, and is usually used for both food and medicine. There were already manyreports about its essential oil and flavonoids compounds, however, the studies aboutthe structure and bioactivities of polysaccharides were still in blank both in domesticand abroad. Polysaccharides were isolated and purified from this plant, and theirprimary structures, antioxidant activities and immunomodulatory effects wereinvestigated systematically in this study. The results could provide theoretical basis ofpharmacologic action of Mosla chinensis Maxim. cv. jiangxiangru, and theexperimental data could also help promote the development and utilization of thisplant.The second part of this research was about the primary structure information(monosaccharide composition, the linkage patterns and sequence of sugar residues) ofhydrolysates from Acacia seyal and Acacia senegal after7hours hydrolysis by0.1MTFA at100℃. Their emulsifying properties of these two Arbic gums were bothimproved by Maillard reaction. This research helped improve the commercial valueand expand its application space.The main research contents and conclusions were summarized as follows,1. On the basis of single factor experiments, response surface methodology(RSM) was used to optimize the extraction conditions of crude polysaccharides fromMosla chinensis Maxim. cv. jiangxiangru (CMP). Three independent variables,extraction temperature, extraction time and water to solid ratio were investigated.Based on the RSM analysis, the optimal extraction conditions were determined asextraction temperature of86.9℃, extraction time of4.1h and water to solid ratio of17.7:1(mL/g). The predictive value of optimal extraction yield was3.38%, which wasconsistent with the real value of3.41%. Protein of CMP was removed by Sevagmethod, fine polysaccharide (MP) was obtained. A neutral polysaccharide MP-1wasisolated and purified by DEAE-52cellulose chromatography column and SephacrylTMS-400HR column. If MP was purified by loading on a Q SepharoseTMFast Flow ionexchange column eluted with1M NaCl and gradient precipitation with ethanol, awater-soluble pectic polysaccharide MP-A40was obtained. 2. The neutral sugar contents of crude polysaccharide (CMP), finepolysaccharide (MP), neutral polysaccharide (MP-1) and acidic polysacchairde(MP-A40) from Mosla chinensis Maxim. cv. jiangxiangru were37.84±0.04%,33.96±0.31%,71.86±0.22%and10.30±0.04%, respectively. Uronic acid content of themwere5.18±0.16%,11.00±0.24%,0%,68.63±0.06%. Potein contents were6.14±0.01%,9.05±0.04%,1.96±0.08%,0.17±0.00%. Monosaccahride compositions ofCMP determined by gas chromatograph were Rha, Rib, Fuc, Ara, Xyl, Man, Glc andGal with a molar ratio of3.72:2.45:0.92:6.00:2.76:5.09:13.53:9.58.Monosaccahride compositions of MP were Rha, Ara, Xyl, Man, Glc and Gal with amolar ratio of5.364:12.260:3.448:12.260:32.567:30.651. Monosaccahridecompositions of MP-1were Glc, Gal, Ara, Man and Rha with a molar ratio of16.02:10.01:4.92:4.62:2.23, and trace amounts of Fuc. MP-A40was composed with4.94%Ara,3.07%Gal,2.13%Rha,1.62%Man and1.29%Glc and68.63%galacturnic acid. The results from HPLC indicated that MP was consisted with at leastfour fractions, and their average molecular weights were4522723Da,254918Da,127350Da and18664Da, repectively. The average molecular weights of MP-1andMP-A40determined by HPSEC were587530Da and32600Da. MP-1and MP-A40were uniform pure polysaccharides determniend by HPSEC method.3. The linkage patterns of MP-1determined by methylation analysis includedâ†'4-Glcp1â†'(31.76%),â†'2-Rhap1â†'(17.11%), t-Galp1â†'(11.02%),â†'2,4-Rhap1â†'(6.47%),â†'3,4-Galp1â†'(5.05%), t-Rhap1â†'(4.31%),t-Glcp1â†'(4.04%), t-Araf1â†'(4.01%),â†'4-Manp1â†'(3.61%),â†'5-Araf1â†'(3.10%),â†'3-Galp1â†'(3.05%),â†'2,4-Manp1â†'(2.55%), t-Fucp1â†'(2.39%) andâ†'4,6-Glcp1â†'(1.53%). The backbone was made ofâ†'4-Glcp1â†'，â†'2-Rhap1â†'，â†'2,4-Rhap1â†', and the main terminal residue was t-Galp1â†'.4. The primary structure of repeat unit of MP-A40wasâ†'4)-[α-GalpA6Me-(1â†']m-[4-α-GalpA-(1â†']n, and the degree of esterification (DE)of MP-A40was32%determined by FTIR method.5. The antioxidant activities in vitro of neutral polysaccharide MP-1and acidicpolysaccharide MP-A40were investited, including DPPH free radical scavengingactivity, ferric ion reducing antixodant power, reducing power and β-carotene-linoleicacid system. The result indicated that MP-1and MP-A40possessed certainantioxidant activities. What’s more, MP-A40had better antoxidant activities.6. The immunosuppressive mice model was built by injecting cyclophosphamide (CY), a common used chemotherapy drug. The restorations of various kinds of relatedenzymes were investigated after feeding the mice by gavag method for a period. Thetissue homogenate was made by granding liver, heart or kidney with0.1g/mL icenormal saline, respectively. And then the homogenate was centrifuged under3000rpm at4°C for10min. The supernatant was used to detect total antioxidant (T-AOC),malondialdehyde (MDA), catalase (CAT), superoxide dismutase (SOD) andglutathione peroxidase (GSH-PX), and other biochemical indicators. The resultsindicated that the spleen and thymus indices and the levels of T-AOC, CAT, SOD, andGSH-PX were raised by MP significantly. However, the MDA level was decreasedunder the same condition. The results proved that MP was able to overcome theimmunosuppression incuced by cyclophosphamide. It indicated that MP might play avery important role during the anti-oxidative damage process in the immun system.7. The capability of the original generation of macrophage’s phagocytosis wasdetected by using neutral red method. As the concentrations of crude polysaccharideCMP increased, the phagocytosis of macrophage was also enhanced. CMP couldstimulate the original generation of peritoneal macrophages to produce NO. As theconcentrations increased, the NO production was increased in a dose-dependentmanner. Spleen lymphocyte proliferation assay indicated that MP-1promoted theproliferation of both T lymphcytes and B lymphcytes. What’s more, MP-1was morelikely to promote T lymphcytes proliferation. The inhibition rate was31.32%whenthe concentration of MP-A40was500μg/mL. The NO production was15times ofthat of the negative control, when MP-A40was of10μg/mL. The NO was from32.58μM to35.02μM when MP-A40was of200μg/mL to500μg/mL. The result indicatedthat MP-A40could increase NO production from RAW264.7.8.2g Acacia seyal or Acacia senegal was dissolved and hydrolyzed in200mL0.1M TFA for7h at100°C. The primary structure informations of hydrolysates wereinvestigated. Methylation analysis indicated that the linkage patterns and their ratiosof Acacia seyal were t-Galp (13.76%),3-Galp (9.70%),4-GalpA (13.70%),6-Galp(30.98%),4,6-Galp (1.63%),3,6-Galp (27.54%),3,4,6-Galp (1.40%) and2,3,6-Galp9(1.27%), and for Acacia Senegal, they were t-Galp (11.65%),â†'3-Galp1â†'(12.41%)，â†'6-Galp1â†'(33.85%),â†'3,4-Galp1â†'(1.34%),â†'4,6-Galp1â†'(1.75%),â†'3,6-Galp1â†'(34.61%),â†'2,6-Galp1â†'(1.14%),â†'2,3,6-Galp1â†'(3.24%). Based on1D/2D NMR (1H,13C, COSY, TOCSY, HSQC, HMBC, NOESY) analysis, thesequence of sugar residues and the structure information of repeating unit were obtained. The backbone was determined to be1,6-Galp with branch linked at O-2，O-3and O-4positions.9. The emulsifying properties of Acacia seyal and Acacia senegal were improvedby Maillard reaction. The reaction was occurred in a closed environment withtemperature of60°C and humidity of75%providing by saturated salt water. Theresult indicated that when the reaction time was7days, the particle size of theemulsion (10%canola oil, v/v) was minimal, and the stability of emulsion was thebest. |
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