發(fā)布時間：2018-05-03 12:57

  本文選題：蛹蟲草基質(zhì)多糖 + 除蛋白�。� 參考：《南京師范大學(xué)》2015年碩士論文

【摘要】：蛹蟲草基質(zhì)多糖是優(yōu)良的保健調(diào)節(jié)劑,能提高機體的免疫、抗氧化功能,并且對正常細(xì)胞無毒副作用。近年來,人們已發(fā)現(xiàn)多種真菌多糖的生物活性與其結(jié)構(gòu)和溶液中的分子鏈構(gòu)象有密切關(guān)系。納米硒是一種高效低毒的優(yōu)于無機硒和有機硒的硒形態(tài),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(yī)學(xué)、生物、化學(xué)等多學(xué)科鄰域研究的焦點和前沿。我們以廢棄的蛹蟲草培養(yǎng)基質(zhì)為原料,從中提取多糖,作為穩(wěn)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(jìn)行初步探索。1.首先研究了蛹蟲草基質(zhì)多糖的高效提取,通過復(fù)合酶法(纖維素酶、果膠酶)協(xié)同超聲波法從廢棄的蛹蟲草培養(yǎng)基質(zhì)中提取基質(zhì)多糖,對提取工藝進(jìn)行優(yōu)化。利用Box-benhnken中心組合試驗設(shè)計優(yōu)化基質(zhì)多糖的提取工藝,建立了復(fù)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(fù)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(zhì)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(biāo),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(yǎng)基中提取的蟲草基質(zhì)粗多糖蛋白的效果。結(jié)果表明：木瓜蛋白酶-TCA法脫蛋白效果最好。粗多糖經(jīng)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(zhì)多糖(CMSP)。3.為了得到理想形貌和粒徑并能夠穩(wěn)定保存的蛹蟲草基質(zhì)多糖納米硒復(fù)合物(CMSP-SeNPs),以提取出的蛹蟲草基質(zhì)多糖為調(diào)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(wěn)定性為指標(biāo),進(jìn)行單因素實驗。確定了在蛹蟲草基質(zhì)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(zhì)多糖納米硒復(fù)合物,以分散均勻的球形顆粒存在,粒徑均一,分散性好,穩(wěn)定性高,基質(zhì)多糖和納米硒粒子是以較弱的物理吸附方式結(jié)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(xì)胞株Hela.乳腺癌細(xì)胞株MCF-7和人肺腺癌細(xì)胞株A549的細(xì)胞毒作用為指標(biāo),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(jié)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(dá)到最高清除率時濃度較��；CMSP-SeNPs對Hela、MCF-7和A549癌細(xì)胞均具有一定抑制率,且對細(xì)胞毒作用呈現(xiàn)出劑量依賴效應(yīng),最小粒徑CMSP-SeNPs(d)的抑制作用也最為明顯,但其對癌細(xì)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