海洋微藻的活性筛选及活性成分的功能研究
发布时间：2019-01-09 07:08
【摘要】：近年来,随着陆地资源的日益减少,人类将眼光转向海洋资源。其中,海洋微藻由于其特殊的生存环境,所以会产生结构新颖和功能独特的活性物质,有很好的药用价值,形成了海洋经济增长点。海洋微藻产业的崛起,为微藻多糖的生产提供了丰富的原材料。本论文以十种常见微藻——杜氏盐藻、集胞藻6803、三角褐指藻、发状念珠藻、螺旋藻、鱼腥藻7120、石耳念珠藻、铜绿微囊藻905、栅藻、海水小球藻(以下简称小球藻)作为研究对象,提取其活性物质进行抑菌实验,初步筛选出具有抑菌作用的微藻。结果显示螺旋藻和小球藻显示出抑菌效果。对筛选出的两种微藻进行粗多糖的进一步研究。螺旋藻和小球藻藻粉热水浸提,离心取上清初步得含较多杂质的粗多糖溶液。经过一系列除杂质蛋白,醇沉,透析,冷冻干燥后得到粗多糖固体产物。粗多糖依次经过纤维素DEAE-52阴离子层析和Sephadex G-200凝胶柱层析分离后得到较纯的螺旋藻多糖。利用苯酚硫酸法测定提取的螺旋藻多糖含糖量,计算得螺旋藻多糖的粗多糖和纯多糖的提取率分别为：2%和83.3%。经过紫外可见光谱扫描,在260nm和280nm处均未出现吸收峰,说明提取的螺旋藻多糖组分中不含有核酸和蛋白质杂质。经过傅里叶红外光谱扫描,测定其有效作用基团。按比例组合小球藻多糖和螺旋藻多糖,利用抑菌及抗氧化活性实验探索复合多糖的比例。复合多糖都对大肠杆菌有抑制作用,其中小球藻多糖质量：螺旋藻多糖质量=1：4时,抑菌效果最好；小球藻和螺旋藻的复合多糖对八叠球菌的抑制作用不明显；小球藻和螺旋藻的复合多糖对假丝酵母均有抑制作用,其中抑菌效果明显的是小球藻多糖：螺旋藻多糖=1：1和1：6达到最好的抑菌效果。小球藻多糖：螺旋藻多糖=1：2时,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的抗脂质过氧化能力最好。螺旋藻多糖：小球藻多糖=1：2时,清除羟基自由基效果最好；小球藻多糖：螺旋藻多糖=1：3时,清除超氧阴离子效果最好；小球藻多糖：螺旋藻多糖=1：3时,其还原力的表现优于其他组。
[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.
【学位授予单位】：天津科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：Q949.2
[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.
【学位授予单位】：天津科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：Q949.2
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