發(fā)布時間：2019-01-09 07:08

【摘要】：近年來,隨著陸地資源的日益減少,人類將眼光轉(zhuǎn)向海洋資源。其中,海洋微藻由于其特殊的生存環(huán)境,所以會產(chǎn)生結(jié)構(gòu)新穎和功能獨(dú)特的活性物質(zhì),有很好的藥用價(jià)值,形成了海洋經(jīng)濟(jì)增長點(diǎn)。海洋微藻產(chǎn)業(yè)的崛起,為微藻多糖的生產(chǎn)提供了豐富的原材料。本論文以十種常見微藻——杜氏鹽藻、集胞藻6803、三角褐指藻、發(fā)狀念珠藻、螺旋藻、魚腥藻7120、石耳念珠藻、銅綠微囊藻905、柵藻、海水小球藻(以下簡稱小球藻)作為研究對象,提取其活性物質(zhì)進(jìn)行抑菌實(shí)驗(yàn),初步篩選出具有抑菌作用的微藻。結(jié)果顯示螺旋藻和小球藻顯示出抑菌效果。對篩選出的兩種微藻進(jìn)行粗多糖的進(jìn)一步研究。螺旋藻和小球藻藻粉熱水浸提,離心取上清初步得含較多雜質(zhì)的粗多糖溶液。經(jīng)過一系列除雜質(zhì)蛋白,醇沉,透析,冷凍干燥后得到粗多糖固體產(chǎn)物。粗多糖依次經(jīng)過纖維素DEAE-52陰離子層析和Sephadex G-200凝膠柱層析分離后得到較純的螺旋藻多糖。利用苯酚硫酸法測定提取的螺旋藻多糖含糖量,計(jì)算得螺旋藻多糖的粗多糖和純多糖的提取率分別為：2%和83.3%。經(jīng)過紫外可見光譜掃描,在260nm和280nm處均未出現(xiàn)吸收峰,說明提取的螺旋藻多糖組分中不含有核酸和蛋白質(zhì)雜質(zhì)。經(jīng)過傅里葉紅外光譜掃描,測定其有效作用基團(tuán)。按比例組合小球藻多糖和螺旋藻多糖,利用抑菌及抗氧化活性實(shí)驗(yàn)探索復(fù)合多糖的比例。復(fù)合多糖都對大腸桿菌有抑制作用,其中小球藻多糖質(zhì)量：螺旋藻多糖質(zhì)量=1：4時,抑菌效果最好；小球藻和螺旋藻的復(fù)合多糖對八疊球菌的抑制作用不明顯；小球藻和螺旋藻的復(fù)合多糖對假絲酵母均有抑制作用,其中抑菌效果明顯的是小球藻多糖：螺旋藻多糖=1：1和1：6達(dá)到最好的抑菌效果。小球藻多糖：螺旋藻多糖=1：2時,清除DPPH自由基效的最好；小球藻多糖：螺旋藻多糖=1：2和螺旋藻多糖：小球藻=1：0的抗脂質(zhì)過氧化能力最好。螺旋藻多糖：小球藻多糖=1：2時,清除羥基自由基效果最好；小球藻多糖：螺旋藻多糖=1：3時,清除超氧陰離子效果最好；小球藻多糖：螺旋藻多糖=1：3時,其還原力的表現(xiàn)優(yōu)于其他組。
[Abstract]:In recent years, with the decreasing of land resources, human beings turn their eyes to marine resources. Because of its special living environment, marine microalgae have a novel structure and unique function of active substances, which has good medicinal value and has formed a marine economic growth point. The rise of marine microalgae industry provides abundant raw materials for the production of microalgae polysaccharides. In this paper, ten kinds of common microalgae, Dunaliella Salina, Chlorophyta 6803, Phaeopsis tricornutum, Candida tricornutum, Spirulina, Anabaena 7120, Candida albicans, Microcystis aeruginosa 905, Chlorophyta, were studied. Chlorella seawater (Chlorella vulgaris) as the research object, extract its active substances for bacteriostasis experiment, the preliminary screening of microalgae with bacteriostasis. The results showed that Spirulina and Chlorella showed bacteriostatic effect. The crude polysaccharides of two microalgae were further studied. Spirulina and Chlorella powder were extracted by hot water and centrifuged to obtain crude polysaccharide solution containing more impurities. After a series of impurity removal, alcohol precipitation, dialysis, freeze-drying, crude polysaccharide solid products were obtained. The crude polysaccharides were separated by cellulose DEAE-52 anion chromatography and Sephadex G-200 gel column chromatography respectively. The sugar content of the polysaccharide extracted from Spirulina platensis was determined by phenol sulfuric acid method. The extraction rates of crude polysaccharide and pure polysaccharide were calculated to be 2% and 83.3% respectively. There were no absorption peaks in 260nm and 280nm, which indicated that the extracted polysaccharides from Spirulina platensis contained no nucleic acid or protein impurities. The effective groups were determined by Fourier transform infrared scanning. Chlorella vulgaris polysaccharides and spirulina polysaccharides were combined in proportion to explore the proportion of composite polysaccharides by bacteriostasis and antioxidant activity experiments. The compound polysaccharides had inhibitory effect on Escherichia coli, and Chlorella platensis polysaccharides had the best inhibitory effect when the mass of polysaccharide was 1:4, the inhibitory effect of the compound polysaccharides of Chlorella vulgaris and Spirulina was not obvious. The compound polysaccharides of Chlorella vulgaris and Spirulina had inhibitory effect on Candida cerevisiae, and the bacteriostatic effect was obvious: the polysaccharide of Chlorella platensis was 1:1 and 1:6, and the best bacteriostatic effect was achieved. Chlorella polysaccharide: spirulina polysaccharide = 1:2, the best scavenging effect of DPPH free radical, Chlorella polysaccharide: spirulina polysaccharide = 1:2 and spirulina polysaccharide: Chlorella vulgaris = 1:0, the ability of anti-lipid peroxidation was the best. Spirulina polysaccharide: Chlorella vulgaris polysaccharide = 1:2, the best scavenging effect of hydroxyl radical, Chlorella platensis polysaccharide: spirulina polysaccharide = 1:3, the best scavenging effect of superoxide anion; Chlorella polysaccharide: spirulina polysaccharide = 1:3, its reducing capacity was better than other groups.
【學(xué)位授予單位】：天津科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：Q949.2

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