本文選題：粒毛盤菌多糖結(jié)構(gòu) + 分子修飾��； 參考：《合肥工業(yè)大學(xué)》2017年碩士論文

【摘要】：本文研究了粒毛盤菌YM240胞外多糖的分離純化、結(jié)構(gòu)表征、羧甲基化修飾、硫酸化修飾,以及修飾前后的體外抗氧化和降血糖及對體內(nèi)降血糖和降血脂活性。粒毛盤菌YM240發(fā)酵液經(jīng)水提醇沉、脫色和脫蛋白后獲得胞外多糖,再經(jīng)DEAE-cellulose 52和Sephadex G-100色譜柱分離純化,得單一組分LEP;HPLC、GC-MS、甲基化分析、FT-IR和1D/2D NMR等實驗結(jié)果表明LEP的分子量為1.68×103 kDa,由摩爾比為16.3:1.0的甘露糖(Man)和半乳糖(Gal)構(gòu)成;(1→3)-α-D-Manp、(1→3,4)-α-D-Manp和(1→3)-α-D-Galp構(gòu)成LEP的主鏈;α-D-Manp-(1→2)-α-D-Manp-(1→3)-α-D-Manp-(1-構(gòu)成支鏈,連接在主鏈上Manp的C-4位。對LEP進行化學(xué)修飾,獲得取代度為0.360的羧甲基化多糖CLEP和取代度為0.144的硫酸化多糖SLEP。紅外光譜和核磁共振碳譜分析結(jié)果表明,多糖的的羧甲基化和硫酸化修飾成功,羧基甲基-CH2COOH取代在(1→2)-α-D-Manp的C-3、(1→3)-α-D-Manp的C-4和(1→)-α-D-Manp的C-6上,而硫酸基-SO3H主要取代在(1→2)-α-D-Manp的C-4和(1→3)-β-D-Galp的C-6上。體外抗氧化實驗表明,與LEP相比,CLEP和SLEP對DPPH自由基和羥基自由基的清除作用及還原力都顯著增強。體外降血糖活性實驗結(jié)果顯示,CLEP和SLEP對α-葡萄糖苷酶和α-淀粉酶抑的制活性及葡萄糖擴散的抑制作用明顯高于LEP。采用不同含量的LEP、CLEP和SLEP處理鏈脲佐菌素(STZ)及高脂飼料誘導(dǎo)的II型糖尿病小鼠(T2DM)。與LEP相比,CLEP和SLEP顯著增加體重、臟器指數(shù)、肝糖原、葡萄糖耐量,明顯降低空腹血糖水平(FBG)、空腹血清胰島素(FINS)和糖化血紅蛋白(HbA1c)以及血清甘油三酯(TG)、膽固醇(TC)和游離脂肪酸(FFA),CLEP的作用更顯著。此外,CLEP和SLEP比LEP更明顯地上調(diào)肝臟中的葡萄糖激酶(GK)和腺苷一磷酸激活蛋白激酶(AMPK),骨骼肌中的AMPK和葡萄糖轉(zhuǎn)運蛋白4(GLUT4),脂肪組織過氧化物酶體增殖物激活受體(PPAR-γ),下調(diào)肝臟中葡萄糖-6-磷酸酶(G6P),并且CLEP比SLEP作用更明顯。因此,CLEP和SLEP具有顯著的降血糖和降血脂活性。
[Abstract]:In this paper, the isolation and purification, structure characterization, carboxymethylation modification, sulfation modification of extracellular polysaccharide of YM240 were studied. The in vitro antioxidation and hypoglycemia, hypoglycemic and hypolipidemic activities were studied before and after modification. Extracellular polysaccharides were obtained from YM240 fermentation broth by water extraction and alcohol precipitation, decolorization and deproteinization, and then purified by DEAE-cellulose 52 and Sephadex G-100 chromatographic column. A single component, LEPP-HPLCX GC-MS.Methylation analysis of FT-IR and 1D/2D NMR showed that the molecular weight of LEP was 1.68 脳 103kDa.The molecular weight of LEP was 1.68 脳 103kDa. the molecular weight of LEP was 1.68 脳 103kDa.The molecular weight of LEP was 16.3: 1.0 mannose) and galactose-a-D-Manp34- 偽 -D-Manp formed the main chain of LEP, 偽 -D-Manp-Manp-1- 偽 -D-Manp-1,3kDa-偽 -D-Manp-131- formed the branching chain of LEP. The C-4 bit of Manp attached to the main chain. Carboxymethylated polysaccharide (CLEP) with a degree of substitution of 0.360 and a sulfated polysaccharide with a degree of substitution of 0.144 were obtained by chemical modification of LEP. The results of IR and NMR showed that carboxymethylation and sulfation of polysaccharides had been successfully modified, and carboxymethyl CH2COOH was substituted on C-4 and C-6 of C-3DMAP and 偽 -D-Manp. The sulfate-SO3H was mainly substituted on C-4 and C-6 of 偽 -D-Manp and 尾 -D-Galp. The antioxidant activity of CLEP and SLEP on DPPH radical and hydroxyl radical was significantly enhanced compared with LEP in vitro. The results of hypoglycemic activity test in vitro showed that CLEP and SLEP inhibited the inhibitory activity of 偽 -glucosidase and 偽 -amylase and the inhibitory effect of glucose diffusion on 偽 -glucosidase and 偽 -amylase were significantly higher than that of LEP. Streptozotocin (STZ) and type II diabetic mice induced by high fat diet were treated with different contents of LEPP-CLEP and SLEP. Compared with LEP, CLEP and SLEP significantly increased body weight, organ index, liver glycogen, glucose tolerance, The effects of FBGN, fasting serum insulin (fins) and glycosylated hemoglobin (HbA1c), serum triglyceride triglyceride (TGN), cholesterol triglyceride (TC) and free fatty acid (FFAA) CLEP were significantly decreased. In addition, CLEP and SLEP upregulated the levels of glucokinase (GK) and adenosine monophosphate activated protein kinase (AMPK) in the liver, AMPK and glucose transporter 4 (GLUT4) in skeletal muscle, adipose tissue peroxisome proliferator activator receptor (PPAR- 緯), PPAR- 緯, respectively. The glucose-6-6 phosphatase G 6 PX in the liver was regulated, and the effect of CLEP was more obvious than that of SLEP. Therefore, CLEP and SLEP have significant hypoglycemic and hypolipidemic activities.
【學(xué)位授予單位】：合肥工業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：O636.1

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