本文選題：蛹蟲草基質多糖 + 除蛋白　； 參考：《南京師范大學》2015年碩士論文

【摘要】：蛹蟲草基質多糖是優良的保健調節劑,能提高機體的免疫、抗氧化功能,并且對正常細胞無毒副作用。近年來,人們已發現多種真菌多糖的生物活性與其結構和溶液中的分子鏈構象有密切關系。納米硒是一種高效低毒的優于無機硒和有機硒的硒形態,具有獨特的光電功能以及良好的生物活性,已成為材料、電子、醫學、生物、化學等多學科鄰域研究的焦點和前沿。我們以廢棄的蛹蟲草培養基質為原料,從中提取多糖,作為穩定劑和包覆試劑,在比較溫和的氧化還原體系中制備硒納米粒子,并對其生物活性進行初步探索。1.首先研究了蛹蟲草基質多糖的高效提取,通過復合酶法(纖維素酶、果膠酶)協同超聲波法從廢棄的蛹蟲草培養基質中提取基質多糖,對提取工藝進行優化。利用Box-benhnken中心組合試驗設計優化基質多糖的提取工藝,建立了復合酶添加量、酶解-超聲溫度、pH值、酶解-超聲時間的四因素回歸模型,確定了最佳提取工藝為在料液比為1：30的條件下,復合酶添加量3%、酶解-超聲溫度54℃、pH值6.0、酶解-超聲時間50mmin的最佳提取條件下,蛹蟲草基質多糖得率為26.38%。2.以蛋白脫除率、多糖損失率和清除DPPH自由基的能力為指標,比較Sevag法,三氯乙酸(TCA)法,木瓜蛋白酶-Sevag法以及木瓜蛋白酶-TCA法脫除從蛹蟲草的廢棄固體培養基中提取的蟲草基質粗多糖蛋白的效果。結果表明：木瓜蛋白酶-TCA法脫蛋白效果最好。粗多糖經木瓜蛋白酶處理過后,TCA法脫除蛋白1次時,蛋白脫除率為92.90%,多糖損失率為9.82%,清除DPPH自由基能力為55.20%,得到蛹蟲草基質多糖(CMSP)。3.為了得到理想形貌和粒徑并能夠穩定保存的蛹蟲草基質多糖納米硒復合物(CMSP-SeNPs),以提取出的蛹蟲草基質多糖為調控劑和修飾劑,以納米硒粒子的粒徑分布、平均粒徑、分散性和穩定性為指標,進行單因素實驗。確定了在蛹蟲草基質多糖濃度為10mg/mL,混合時間為4h,亞硒酸濃度為1.0M,滴加速度為1.5rpm的條件下時,能夠制備出含硒量為54.8mg/L的蛹蟲草基質多糖納米硒復合物,以分散均勻的球形顆粒存在,粒徑均一,分散性好,穩定性高,基質多糖和納米硒粒子是以較弱的物理吸附方式結合在一起。4.以對DPPH和羥自由基的清除能力以及對人宮頸癌細胞株Hela.乳腺癌細胞株MCF-7和人肺腺癌細胞株A549的細胞毒作用為指標,考察了四種粒徑大小不同的CMSP-SeNPs的體外抗氧化和抗腫瘤能力。結果顯示,最小粒徑CMSP-SeNPs(d)對DPPH和羥自由基的清除能力均大于CMSP, Na2Se03,且達到最高清除率時濃度較小；CMSP-SeNPs對Hela、MCF-7和A549癌細胞均具有一定抑制率,且對細胞毒作用呈現出劑量依賴效應,最小粒徑CMSP-SeNPs(d)的抑制作用也最為明顯,但其對癌細胞的抑制率不及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，

本文編號：1838575