松茸多糖的提

发布时间：2018-06-06 18:26

  本文选题：松茸多糖 + 提取和分离　； 参考：《聊城大学》2017年硕士论文

【摘要】：松茸,因其鲜美的味道、嫩滑的肉质,很受人们喜欢,是一种珍稀名贵的纯天然食药用真菌。它含有多糖、氨基酸和蛋白质等成分,且有抗肿瘤、治疗心血管疾病和促进肠胃功能等功效,被誉为“食用菌之王”。松茸多糖是其主要的活性成分之一,具有较高的营养价值和药用活性,松茸多糖在保健品和医药等领域具有广阔的开发价值和应用前景。本文设计了5种培养基配方对松茸菌丝体进行固体培养,筛选出最适于松茸生长的固体培养基培养,为松茸菌的人工栽培提供理论依据。采取超声辅助热水浸提法、传统热水浸提法、酶提取法和微波提取法4种提取方式进行松茸多糖的提取,通过对照松茸多糖的得率和抗氧化活性,筛选出最优的提取方式为超声辅助热水浸提法。通过单因素和正交试验设计筛选出超声辅助热水浸提法的最优提取工艺,进而采取乙醇分级的方法对松茸多糖进行分离纯化,并测定各组分的单糖成分和抗氧化活性,得到更有利于生物医学发展的可能有效成份,为更深地研究松茸多糖提供科学依据。研究结果如下:1.研究和比较了5种培养基的生长状况,根据菌丝体的长势(菌丝密度、菌丝形态,菌丝颜色、菌丝边缘整齐度)和日生长速率,筛选出适合松茸母种的最优培养基,配方为:马铃薯葡萄糖琼脂培养基(PDA)+蛋白胨+麦麸汁+松针汁(即:马铃薯20 g、葡萄糖2 g、琼脂2 g、蒸馏水100 m L、蛋白胨0.3 g、麦麸汁3 g、松针汁3 g)。2.对4种提取方式进行了比较:热水浸提法(H-TMP)、超声辅助提取法(U-TMP)、微波提取法(U-TMP)和酶提取法(E-TMP),4种方法的提取多糖提取得率和DPPH自由基的清除力高低顺序均为:H-TMPU-TMPE-TMPM-TMP,它们的还原力的高低顺序为:U-TMPH-TMPM-TMPE-TMP;羟自由基的清除力顺序为:U-TMPH-TMPE-TMPM-TMP。综合考虑4种提取方法提取的多糖得率和抗氧化活性,可以看出超声提取法的得率和活性相对较高,而且,与传统的热水浸提法相比较,可以节省时间,提高效率。3.采用超声辅助法制备松茸多糖,并采取单因素和正交试验设计((L9(3)3)对超声辅助法的工艺条件进行了优化。结果显示,超声辅助法的最优提取工艺为:超声温度40℃,超声时间50 min,水料比例25 m L/g,超声频率45 k Hz,超声功率100W,提取得率为8.06%。4.通过乙醇分级法从松茸多糖中分离纯化出3种新颖的多糖(TMP30、TMP60和TMP80),采用高效阴离子交换色谱法(HPAEC-PAD)和傅立叶变换红外光谱法(FT-IR)进行了结构表征。抗氧化活性是研究其还原力、DPPH自由基清除能力和羟自由基清除能力三方面。3种多糖的抗氧化活性高低顺序均为:TMP80TMPTMP60TMP30。由此可以看出,松茸多糖分离纯化得到的3种多糖,具有一定的抗氧化活性,尤其是TMP80,是一种有很大开发前景的组分。
[Abstract]:Matsutake, because of its delicious taste, smooth meat, very popular with people, is a rare and rare natural edible medicinal fungi. It contains polysaccharides, amino acids and proteins, and has the function of anti-tumor, treating cardiovascular diseases and promoting the function of stomach and intestines. It is called "King of Edible Fungi". Tricholoma matsutake polysaccharide is one of its main active components and has high nutritional value and medicinal activity. It has broad development value and application prospect in health products and medicine and other fields. In this paper, five kinds of medium formulations were designed for solid culture of Tricholoma matsutake mycelium, and the best solid medium for the growth of Tricholoma matsutake was selected, which provides theoretical basis for the artificial cultivation of Tricholoma matsutake. The polysaccharides of Tricholoma matsutake were extracted by ultrasonic assisted hot water extraction, traditional hot water extraction, enzyme extraction and microwave extraction. The extraction rate and antioxidant activity of polysaccharides from Tricholoma matsutake were compared. The best extraction method is ultrasonic assisted hot water extraction. The optimum extraction process of ultrasonic assisted hot water extraction method was screened by single factor and orthogonal design, and then the polysaccharide of Tricholoma matsutake was separated and purified by ethanol classification, and the monosaccharide composition and antioxidant activity of each component were determined. The possible active components which are more favorable to the development of biomedicine can be obtained, which provides scientific basis for the further study of Tricholoma matsutake polysaccharides. The results are as follows: 1. The growth status of five kinds of media was studied and compared. According to the growth of mycelium (mycelium density, mycelium morphology, hypha color, hyphal edge uniformity) and daily growth rate, the optimum medium for matsutake mother strain was selected. The formula is: potato glucose Agar medium (PDAA) peptone wort juice (i.e. potato 20 g, glucose 2 g, Agar 2 g, distilled water 100 mL, peptone 0.3 g, wort juice 3 g, pine needle juice 3 g 路L ~ (-2), the formula is: potato 20 g, glucose 2 g, Agar 2 g, distilled water 100 mL, peptone 0.3 g, wort juice 3 g, pine needle juice 3 g 路2. The extraction rate of polysaccharides and the scavenging power of DPPH free radical of four methods were compared: hot water extraction, ultrasonic assisted extraction, microwave-assisted extraction, microwave extraction and enzyme extraction. The order of their extraction rates and scavenging power of DPPH free radicals were respectively: (1) H-TMPU-TMPE-TMPM-TMP2, and (2) the order of scavenging activity of DPPH free radical was: (1) H-TMPU-TMPM-TMPM-TMP. the order of extraction of polysaccharides and the scavenging power of DPPH free radical were: The order of reducing force is: U -TMPH-TMPM-TMPE-TMPE-TMPand the order of scavenging power of hydroxyl radical is: U U-TMPH-TMPE-TMPM-TMP. Considering the extraction rate of polysaccharides and antioxidant activity of the four extraction methods, it can be seen that the extraction rate and activity of ultrasonic extraction method are relatively high, and compared with the traditional hot water extraction method, it can save time and improve efficiency. The polysaccharides of Tricholoma matsutake were prepared by ultrasonic assisted method, and the technological conditions of the ultrasonic assisted method were optimized by single factor and orthogonal design. The results showed that the optimum extraction process was as follows: ultrasonic temperature 40 鈩，

本文编号：1987693