蛹虫草基质多糖纳米硒复合物的制备及其生物活性初探

发布时间：2018-05-03 12:57

  本文选题：蛹虫草基质多糖 + 除蛋白　； 参考：《南京师范大学》2015年硕士论文

【摘要】：蛹虫草基质多糖是优良的保健调节剂,能提高机体的免疫、抗氧化功能,并且对正常细胞无毒副作用。近年来,人们已发现多种真菌多糖的生物活性与其结构和溶液中的分子链构象有密切关系。纳米硒是一种高效低毒的优于无机硒和有机硒的硒形态,具有独特的光电功能以及良好的生物活性,已成为材料、电子、医学、生物、化学等多学科邻域研究的焦点和前沿。我们以废弃的蛹虫草培养基质为原料,从中提取多糖,作为稳定剂和包覆试剂,在比较温和的氧化还原体系中制备硒纳米粒子,并对其生物活性进行初步探索。1.首先研究了蛹虫草基质多糖的高效提取,通过复合酶法(纤维素酶、果胶酶)协同超声波法从废弃的蛹虫草培养基质中提取基质多糖,对提取工艺进行优化。利用Box-benhnken中心组合试验设计优化基质多糖的提取工艺,建立了复合酶添加量、酶解-超声温度、pH值、酶解-超声时间的四因素回归模型,确定了最佳提取工艺为在料液比为1：30的条件下,复合酶添加量3%、酶解-超声温度54℃、pH值6.0、酶解-超声时间50mmin的最佳提取条件下,蛹虫草基质多糖得率为26.38%。2.以蛋白脱除率、多糖损失率和清除DPPH自由基的能力为指标,比较Sevag法,三氯乙酸(TCA)法,木瓜蛋白酶-Sevag法以及木瓜蛋白酶-TCA法脱除从蛹虫草的废弃固体培养基中提取的虫草基质粗多糖蛋白的效果。结果表明：木瓜蛋白酶-TCA法脱蛋白效果最好。粗多糖经木瓜蛋白酶处理过后,TCA法脱除蛋白1次时,蛋白脱除率为92.90%,多糖损失率为9.82%,清除DPPH自由基能力为55.20%,得到蛹虫草基质多糖(CMSP)。3.为了得到理想形貌和粒径并能够稳定保存的蛹虫草基质多糖纳米硒复合物(CMSP-SeNPs),以提取出的蛹虫草基质多糖为调控剂和修饰剂,以纳米硒粒子的粒径分布、平均粒径、分散性和稳定性为指标,进行单因素实验。确定了在蛹虫草基质多糖浓度为10mg/mL,混合时间为4h,亚硒酸浓度为1.0M,滴加速度为1.5rpm的条件下时,能够制备出含硒量为54.8mg/L的蛹虫草基质多糖纳米硒复合物,以分散均匀的球形颗粒存在,粒径均一,分散性好,稳定性高,基质多糖和纳米硒粒子是以较弱的物理吸附方式结合在一起。4.以对DPPH和羟自由基的清除能力以及对人宫颈癌细胞株Hela.乳腺癌细胞株MCF-7和人肺腺癌细胞株A549的细胞毒作用为指标,考察了四种粒径大小不同的CMSP-SeNPs的体外抗氧化和抗肿瘤能力。结果显示,最小粒径CMSP-SeNPs(d)对DPPH和羟自由基的清除能力均大于CMSP, Na2Se03,且达到最高清除率时浓度较小；CMSP-SeNPs对Hela、MCF-7和A549癌细胞均具有一定抑制率,且对细胞毒作用呈现出剂量依赖效应,最小粒径CMSP-SeNPs(d)的抑制作用也最为明显,但其对癌细胞的抑制率不及Na2SeO3。
[Abstract]:Cordyceps militaris matrix polysaccharide is an excellent health care regulator, which can improve the immune and antioxidant function of the body, and has no toxic and side effects on normal cells. In recent years, it has been found that the biological activities of various fungal polysaccharides are closely related to their structure and molecular chain conformation in solution. Nano-selenium is a kind of selenium form with high efficiency and low toxicity, which is superior to inorganic selenium and organic selenium. It has unique optoelectronic function and good biological activity. It has become the focus and frontier of multi-disciplinary neighborhood research, such as materials, electronics, medicine, biology, chemistry and so on. Polysaccharides were extracted from waste culture medium of Cordyceps militaris as stabilizers and coated reagents to prepare selenium nanoparticles in a mild redox system and their biological activities were preliminarily explored. Firstly, the extraction of polysaccharide from Cordyceps militaris was studied. The polysaccharide was extracted from the culture matrix of Cordyceps militaris by combined enzymatic method (cellulase, Pectinase) with ultrasonic method, and the extraction process was optimized. The extraction process of matrix polysaccharides was optimized by Box-benhnken center combination test. A four-factor regression model was established, which included the amount of complex enzyme, the pH value of enzymatic hydrolysis and ultrasonic temperature, and the enzymatic hydrolysis and ultrasonic time. The optimum extraction conditions were determined as follows: the optimum extraction conditions were as follows: the ratio of material to liquid was 1:30, the amount of compound enzyme was 3, the pH value of enzyme hydrolyzation-ultrasonic temperature was 54 鈩，

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