【作者】 陈晓清；

【导师】 郑怡；

【作者基本信息】 福建师范大学 ， 植物学， 2005， 硕士

【摘要】 分别从二种微藻一紫球藻和海水小球藻中分离纯化多糖、凝集素和抗菌蛋白,并对它们的一些特性进行了研究,结果如下:1. 紫球藻凝集素（PCL）是一种分子量为32.5 kD³9.6 kD的糖蛋白,分子中含有28.4%的中性糖。在氨基酸组成中,缬氨酸的含量最高,其次是谷氨酸与甘氨酸,不含蛋氨酸、组氨酸、精氨酸和脯氨酸。PCL能凝集兔和猪的红细胞,能凝集4种单细胞藻类,还能够凝集中华根霉和甘薯薯瘟病病原菌以及经热处理的枯草芽孢杆菌和啤酒酵母。PCL的凝血活性可被某些单糖或二糖及胃粘蛋白所抑制,对热敏感,对酸碱处理有较好的稳定性,Ca²⁺、Mn²⁺、Zn²⁺能激活其被EDTA-Na₂所抑制的活性。2. 海水小球藻凝集素（CPL）是一种分子量为63.9 kD-一70.3 kD的糖蛋白,分子中含有10.8%的中性糖。在氨基酸组成中,甘氨酸的含量最高,其次是谷氨酸与天冬氨酸,不含脯氨酸。CPL能凝集兔、山羊、鸡和猪的红细胞,能凝集5种单细胞藻类,还能够凝集大肠杆菌、甘薯薯瘟病病原菌、稻瘟病病原菌以及经热处理的变形杆菌。CPL的凝血活性能被某些单糖或二糖所抑制,对热、酸碱处理有较好的稳定性,Ca²⁺、Mn²⁺、Zn²⁺能激活其被EDTA-Na₂所抑制的活性。3.从紫球藻中得到一种抗菌蛋白,在SDS-PAGE上显示2条带,分子量分别为2l kD与30 kD,氨基酸组成分析表明,此抗菌蛋白含17种氨基酸,尤富含谷氨酸、甘氨酸与天冬氨酸。纯化的蛋白对产黄青霉和甘蔗黑穗病病原菌有较强的抑制作用,对枯草杆菌和藤黄八叠球菌也有抑制作用,抗菌活性与温度变化有关。4.从海水小球藻中得到一种抗菌蛋白,在SDS-PAGE上显示2条带,分子量分别为61 kD与70 kD,氨基酸组成分析表明,此抗菌蛋白含17种氨基更多还原

【Abstract】 Lectin antibiotic protein and polysaccharide were isolated and purified from two microalgae, Porphyridium cruentum and Chlorella pacifica, and were investigate their partial characterizations. The conclusions were as follows:1. Porphyridium cruentum lectin (PCL) was a glycoprotein with a relative molecular weight of 32.5kD~39.6kD, containing 28.4% neutral succharide. In its amino acid composition, Val has the highteat content, followed by Glu and Gly, but there were lack of Met His,. Arg and Pro. PCL could agglutinated erythrocytes from rabbit and pig .Further more, PCL could agglutinated four species of unicellular algae and some bacteria or fungia such as Rhizopus chinensis Saito Fasarium oxyssporam, Bacillus subtilis Cohn and Saccharomyces carlsbergengensis Hansen treated by heating. The agglutinating activity was sensitive to temperature change and stable over a wide pH range, but inhibited by the presence of some mono or bisaccharides and mucin. Inhibition of EDTA-Na2 was activated adding of Ca2+ Mn2+ and Zn2+2. Chlorella pacifica lectin (CPL) was a glycoprotein with a relative molecular weight of 63.9kD~70.kD, containing 10.8% neutral succharide. In its amino acid composition, Gly has the highteat content, followed by Glu and Asp, but there were lack of Pro. CPL could agglutinated erythrocytes from rabbit goat chicken and pig. Further more, CPL could agglutinated five species of unicellular algae and some bacteria or fungia such as Escherichia coli Castellani et Chalmers, Fasarium oxyssporam Piricularia oryzae Cav and Proteas valgaris Hauser treated by heating. The agglutinating activity was insensitive to temperature change and stable over a wide pH range, but inhibited by the presence of some mono or bisaccharides. Inhibition of EDTA-Na2 was activated adding of Ca2+ Mn2+andZn2+3. A antibiotic protein was isolated and purified from Porphyridium cruentum. In SDS-PAGE the protein appears two bands and their molecular weight were 21kD and 30kD, respectively. Analysis of amino acid composition suggests this protein contains of 17 amino acid, especially with high content of Glu> Gly and Asp. The protein was strongly activity against two fungi, Penicillium chrysogenum Thom and Ustilagu Scitaminea Sydow, and exhibited a moderate activity against Bacillus subtilis Cohn and Sarcinalutea Schroeter. This antibiotic protein was sensitive to temperature change.4. A antibiotic protein was isolated and purified from Chlorella pacifica, In SDS-PAGE the protein appears two bands and their molecular weight was 61kD and 70kD, respectively. Analysis of amino acid composition suggests this protein contains of 17 amino acid, especially with high content of Glu> Gly and Asp. The protein was strongly activity against two fungi, Penicillium chrysogenum Thom and Rhizopus chinensis Saito, the protein also exhibited a moderate antibacterial action against Staphylococcus aureus Roenbach and Ameromonas punctata. The protein was insensitive to temperature change. Ingeneral, antibiotic proteins from the two microalgae inhibited antifungal activities more strongly than antibacterial activity.5. Crude polysaccharides from Chlorella pacifica and Porphyridium cruentum were extracted by hot water .followed by deproteinizining(Sevag) and purified by DEAE-cellulose. Extracts obtained were tested the antibiotic activities using the agar disc diffusion method. The result showed that there were few differences between antibacterial activities and antifungal activities of polysaccharide extracts from Porphyridium cruentum, but it has strong activities against Pseudomonas fluorecens and Piricularia oryzae Cav. The minimum inhibitionconcentration (MICs) of polysaccharides against Piricularia oryzae Cav was 6.25 mg/ mL. The polysaccharide extract from Chlorella pacifica had more strong activity fugia than against bacteria. It showed strong activities aainst Rhizopus chinensis Saito Sarcina lutea Schroeter Aspergillus niger V.Tiegh Helminthos porium turcicum Pass and Piricularia oryzae Cav . The minimum inhibition concentration (MICs) of polysaccharides against Piricularia oryzae Cav and Rhizopus chinensis Saito was 6.25 mg/ mL.In a word, Porphyridium cruentum and Chlorella pacifica have richbiologically active compounds, so they have great value of development andutilization.更多还原