板栗多糖的提取、分離純化及抗氧化活性研究

生物工程論文編號:SW095  論文字?jǐn)?shù):12629,頁數(shù):28

目  錄
中文摘要 I
英文摘要 II
目  錄 III
1. 緒 論 1
1.1 板栗的研究概況 1
1.2 多糖的概述及其研究進(jìn)展 2
2  實驗部分 7
2.1 多糖的提取 7
2.2 板栗粗糖蛋白和粗多糖的理化性質(zhì)測定 8
2.3 板栗粗多糖的分離純化 10
2.4 板栗多糖的抗羥基自由基活性測試 11
3  結(jié)果與討論 12
3.1板栗多糖的提取 12
3.2 板栗粗糖蛋白和粗多糖的理化性質(zhì)測定 12
3.3 板栗粗多糖的分離純化 17
3.4 板栗粗多糖對羥基自由基·OH的清除率測定 19
4. 總結(jié)與展望 21
致 謝 22
參考文獻(xiàn) 23

摘要：板栗營養(yǎng)價值豐富，具有養(yǎng)胃健脾，壯腰補腎的藥用價值，國內(nèi)對板栗多糖的研究鮮少報道。本文對去殼板栗經(jīng)乙醇脫脂，熱水浸提，醇析得到粗糖蛋白，并對提取得到的板栗粗糖蛋白進(jìn)行脫蛋白工藝處理，得到板栗粗多糖4.42g，得率為1.77%。同時對板栗糖蛋白和板栗粗多糖進(jìn)行了理化性質(zhì)的測定，粗多糖在280nm處無吸收峰，說明蛋白質(zhì)已基本除盡�？捡R斯亮藍(lán)G-250測得板栗粗糖蛋白中蛋白質(zhì)含量為2.28%，板栗粗多糖中蛋白質(zhì)含量為0.026%。苯酚-硫酸法測得板栗粗多糖中己糖含量為89.54%。紅外光譜顯示，該粗多糖可能含有β-D-葡萄吡喃糖基。
     板栗粗多糖經(jīng)Sephacryl S-100HR柱、DEAE-Sepadax Fast Flow陰離子交換柱層析分離后得到了8個板栗多糖組分。對板栗多糖組分CNPS-8進(jìn)行純度檢驗，采用層析儀經(jīng)Sephacryl S-100HR洗脫，得到了一個單一對稱的峰，證明該組分為一均一的精多糖。
     采用甲基紫模型，對板栗粗多糖進(jìn)行羥基自由基清除率試驗，結(jié)果證明板栗粗多糖在劑量為1-20mg/mL范圍內(nèi),對羥基自由基具有一定的清除效果。且隨濃度的增大，清除效果增強。但在同等劑量下弱于陽性對照Vc，20 mg/mL的板栗粗多糖的清除率為34.8%，而陽性對照Vc(20mg/mL)的清除率為66.08%。
關(guān)鍵詞：板栗；多糖；提取；分離純化；抗氧化活性  
Polysaccharides from Chestnut: Extraction, isolation,
 purification and antioxidant activity

Abstract: Being rich of nutrition, Chestnut has a lot of biological activity and high medicinal function, and it’s benefit to stomach, spleen, spinal and kidney. Domestic research of Chestnut polysaccharide was rarely reported. In this paper, crude polysaccharide is obtained by following process to peeled chestnuts: degreased by ethanol; hot-water extraction; alcohol sedimentation (glycoprotein); deproteinizing, which yielding 1.77% for 4.42g. Meanwhile, characteristics of Chestnut glycoprotein and Chestnut crude polysaccharide are mensurated, it shows that protein has been basically eliminated for there is no absorption peak at 280 nm.  Protein content of glycoprotein and crude polysaccharide are measured by Coomassie Brilliant Blue .which are separately to 2.28% and 0.026% . Hexose content of crude polysaccharide is measured by Phenol - sulfuric acid method to 89.54%. Infrared spectroscopy revealed that the crude polysaccharide may contain Beta-D-based glucopynanosyl.
 By Sephacryl S-100HR column and DEAE - Sepadax Fast Flow anion-exchange column, 8 components are separated from Chestnut crude polysaccharide. A single symmetrical peak is gained by Elution through Sephacryl S-100HR, what proves that the group is divided into a  homogeneous precision polysaccharide.
 Hydroxy free radical scavenging tests for chestnut crude polysaccharide is carried out using methyl violet model.It confirms that Chestnut crude polysaccharide has certain hydroxyl radical scavenging effect range 1-20mg/mL, and the effect begin strong as the bigger of the content of the crude polysaccharide. The scavenging activity on hydroxy free radical is weaker than Vc, the result shows, the scavenging effect of the crude polysaccharide (20mg/mL) is 34.8%, while Vc  (20mg/mL) is 66.08%.
Keywords：Chestnut; Polysaccharide; Extraction; Purification; Antioxidant Activity