本文選題：紅松松子殼多糖 + 分離純化��； 參考：《吉林大學(xué)》2017年碩士論文

【摘要】：多糖又稱多聚糖,由10個(gè)以上單糖通過(guò)糖苷鍵連接而成,廣泛存在于生物體中,在生命活動(dòng)中具有重要作用。多糖具有抗腫瘤、抗病毒、抗氧化和抗菌等生物活性。紅松松子殼多糖(Pinuskoraiensis Pinon Shell Polysaccharides,PPSP)是從中國(guó)東北地區(qū)經(jīng)濟(jì)型林木紅松松子殼廢棄物中提取而來(lái),因其具有較強(qiáng)的抗病毒、抗腫瘤等生物活性而備受關(guān)注。本研究首先通過(guò)超聲輔助熱水浸提法提取PPSP,將純化后的PPSP-b采用離子交換層析和葡聚糖凝膠層析法進(jìn)行分離,然后通過(guò)紫外可見(jiàn)光分光光度計(jì)和化學(xué)方法檢測(cè)PPSP及其主要多糖組分PPSP2-2和PPSP3-1的理化性質(zhì)。本實(shí)驗(yàn)中,超聲輔助熱水浸提法提取PPSP條件:提取溫度為80℃,浸提時(shí)間為40 min,浸提次數(shù)為2次,固液比為1:3。結(jié)果表明,在此條件下多糖提取率為2.37±0.23%,糖含量為25.78%;PPSP經(jīng)過(guò)木瓜蛋白酶結(jié)合Sevag法脫蛋白,蛋白去除率為90.87%,多糖保留率為76.41%;PPSP-b經(jīng)過(guò)DEAE-52離子交換層析后分離得到PPSP1、PPSP2和PPSP3,因PPSP1含量低,本實(shí)驗(yàn)將PPSP2和PPSP3進(jìn)一步經(jīng)過(guò)葡聚糖凝膠層析分別得到PPSP2-1、PPSP2-2和PPSP3-1、PPSP3-2,又因PPSP2-1和PPSP3-2含量較少,最終將PPSP2-2和PPSP3-1作為主要研究對(duì)象。理化性質(zhì)的檢測(cè)結(jié)果表明,PPSP、PPSP2-2、PPSP3-1均以多糖成分為主。然后本研究對(duì)PPSP2-2和PPSP3-1的一級(jí)結(jié)構(gòu)和空間結(jié)構(gòu)進(jìn)行測(cè)定,剛果紅實(shí)驗(yàn)結(jié)果表明,PPSP2-2分子內(nèi)存在三股螺旋構(gòu)象,而PPSP3-1分子內(nèi)不存在三股螺旋構(gòu)象;高效液相色譜圖分析表明,PPSP2-2單糖組成及其摩爾比為Man:Rha:Glc A:Gal A:Glc:Gal:Xyl:Ara:Fuc=7.3:12.1:3.6:11.8:11.0:20.2:10.5:18.4:5.1;PPSP3-1單糖組成及其摩爾比為Man:Rha:Glc A:Gal A:Glc:Gal:Xyl:Ara:Fuc=4.6:5.6:2.9:44.9:23.8:9.1:2.5:5.2:1.3,PPSP2-2和PPSP3-1分子量分布相對(duì)均一,且分子量都較小,分別為1.89×103 Da、2.84×103Da;紅外光譜結(jié)果顯示,PPSP2-2和PPSP3-1均是酸性多糖,且都含有吡喃糖。最后,本研究對(duì)PPSP對(duì)小鼠抗氧化活性的影響進(jìn)行檢測(cè),以不同劑量PPSP對(duì)小鼠連續(xù)灌胃21 d,檢測(cè)小鼠肝臟和血清中的超氧化物歧化酶(super oxide dismutase,SOD)、過(guò)氧化氫酶(catalase,CAT)、谷胱甘肽過(guò)氧化物酶(glutathione peroxidase,GSH-Px)活力和丙二醛(malondialdehyde,MDA)含量。結(jié)果表明,在灌胃不同劑量的PPSP后,小鼠的抗氧化作用均有所提高,且呈現(xiàn)出一定的劑量效應(yīng)。與對(duì)照組相比,PPSP高劑量組(400mg/(kg·d))能極顯著提高小鼠血清和肝臟中的SOD、CAT和GSH-Px活性(P0.01),并極顯著降低MDA含量(P0.01);PPSP中劑量組(200mg/(kg·d))能夠極顯著提高(P0.01)小鼠肝臟組織中的SOD活力,并顯著提高(P0.05)小鼠血清中的GSH-Px活力,同時(shí)顯著降低小鼠血清和肝臟中的MDA含量(P0.05)。綜上所述,本研究通過(guò)分離純化PPSP,得到四種不同多糖組分,其中PPSP2-2和PPSP3-1是其主要組成部分,PPSP2-2主要由Gal、Ara、Rha、Gal A、Glc、Xyl單糖組成,分子量為1.89×103 Da;PPSP3-1主要由Gal A、Glc、Gal等單糖組成,分子量為2.84×103Da;通過(guò)紅外檢測(cè),PPSP2-2和PPSP3-1是含有吡喃糖的酸性多糖。PPSP具有體內(nèi)抗氧化生物活性,且呈現(xiàn)出劑量效應(yīng)。
[Abstract]:Polysaccharide, also called polysaccharide, is made up of more than 10 monosaccharides through glucoside bonds. It is widely found in organisms and plays an important role in life activities. Polysaccharides have biological activities, such as anti-tumor, antiviral, antioxidant and antibacterial. Pinuskoraiensis Pinon Shell Polysaccharides (PPSP) is from the northeast of China. In this study, PPSP was extracted by ultrasonic assisted hot water extraction, and the purified PPSP-b was separated by ion exchange chromatography and glucan gel layer analysis, and then UV visible light was used. The physicochemical properties of PPSP and its main polysaccharide components PPSP2-2 and PPSP3-1 were detected by photometer and chemical method. In this experiment, the ultrasonic assisted hot water extraction method was used to extract the PPSP conditions: the extraction temperature was 80 C, the extraction time was 40 min, the extraction times were 2 times, and the solid-liquid ratio was 1:3.. The extraction rate of polysaccharide was 2.37 + 0.23% and the sugar content under this condition. For 25.78%, PPSP was deproteinized by papain combined with Sevag, the removal rate of protein was 90.87%, the retention rate of polysaccharide was 76.41%, and PPSP1, PPSP2 and PPSP3 were separated after DEAE-52 ion exchange chromatography. Because of low PPSP1 content, PPSP2 and PPSP3 were further obtained by glucan gel chromatography, respectively. PSP3-2, also due to the low content of PPSP2-1 and PPSP3-2, finally took PPSP2-2 and PPSP3-1 as the main research object. The results of physical and chemical properties showed that PPSP, PPSP2-2 and PPSP3-1 were dominated by polysaccharide components. Then the first order structure and spatial structure of PPSP2-2 and PPSP3-1 were measured. The results of Congo red experiment showed that PPSP2-2 molecular memory was in memory. In three strands of spiral conformation, there is no three strand conformation in PPSP3-1, and high performance liquid chromatography analysis shows that the composition of PPSP2-2 monosaccharide and its mole ratio are Man:Rha:Glc A:Gal A:Glc:Gal:Xyl:Ara:Fuc=7.3:12.1:3.6:11.8:11.0:20.2:10.5:18.4:5.1, and the composition of PPSP3-1 monosaccharide and its mole ratio are Man:Rha:Glc A:Gal A:Glc:Gal:Xyl:Ara:Fuc= The molecular weight distribution of 4.6:5.6:2.9:44.9:23.8:9.1:2.5:5.2:1.3, PPSP2-2 and PPSP3-1 is relatively uniform, and the molecular weight is smaller, 1.89 * 103 Da and 2.84 x 103Da respectively. The infrared spectrum shows that both PPSP2-2 and PPSP3-1 are acid polysaccharides and contain Piran. Finally, the study on the effect of PPSP on the antioxidant activity of mice is not used. The same dose of PPSP was administered to mice continuously for 21 d, and the activity of superoxide dismutase (super oxide dismutase, SOD), catalase (catalase, CAT), glutathione peroxidase (glutathione peroxidase, GSH-Px) and the content of malondialdehyde (malondialdehyde, etc.) in the liver and serum of mice were detected. In contrast to the control group, the PPSP high dose group (400mg/ (kg. D)) could significantly increase the SOD, CAT and GSH-Px activity (P0.01) in the serum and liver of mice, and significantly reduced the MDA content (P0.01) in the mice, and the PPSP dose group (200mg/ (200mg/)) could significantly improve the small dose. The activity of SOD in rat liver tissue, and significantly increase the GSH-Px activity in the serum of (P0.05) mice, and significantly reduce the MDA content in the serum and liver of mice (P0.05). To sum up, this study obtained four different polysaccharide components by separating and purifying PPSP, in which PPSP2-2 and PPSP3-1 are its main components, PPSP2-2 mainly from Gal, Ara, Rha, Rha. Al A, Glc, and Xyl monosaccharides are composed of 1.89 x 103 Da; PPSP3-1 is mainly composed of Gal A, Glc, Gal and other monosaccharides, and the molecular weight is 2.84 x 103Da.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R914

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