一种褐藻—铁钉菜（Ishige okamurae）凝集素的研究
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摘要
本研究以褐藻门的铁钉菜(Ishige okamurae)为试验材料,经PBS浸提、硫酸铵沉淀、DEAE-Sepharose FF离子交换层析和Sephadex G-100分子筛层析,分离纯化得到铁钉菜凝集素(IOL),其对兔血红细胞最小凝集浓度约为156.25μg/mL。IOL在SDS-PAGE上显示单一蛋白染色带,测定的相对分子量约为45KDa。
理化特性的研究结果表明,铁钉菜凝集素含有17.877%的中性糖,最大紫外吸收峰为269.3nm;在氨基酸组成中,谷氨酸(Glu)的含量最高,天门冬氨酸(Asp)的含量次之,含量最低的是甲硫(蛋)氨酸(Met),不含色氨酸(Try)。凝集活性只被麦牙糖所抑制。在pH 4.0～9.0范围内相对稳定;具有较高的热稳定性,最高耐热温度为80℃;此外,IOL对二价金属离子Zn~(2+)和Mg~(2+)具有不同程度的依赖性。
生物学活性实验显示,IOL对7种供试真菌(菌丝生长及孢子萌发)及11种细菌生长具有不同程度的影响,能够凝集斜生栅藻、湛江叉鞭金藻和绿色巴夫藻,对供试藻类的生长及叶绿素a含量的影响不同,能抑制多纹膝沟藻和棕囊藻的生长,降低叶绿素a含量,但能够促进亚心型扁藻、斜生栅藻和红色蔷薇藻的生长,使它们叶绿素a的含量提高。
A lectin,named IOL,was isolated and purified from a brown marine alga Ishige okamurae by extraction with PBS,followed by ammonium sulfate precipitation and a combination of ion-exchange chromatography on DEAE-Sepharose fast flow and gel filtration on Sephadex G-100,and the lowest agglutinating concentration of IOL tested with erythrocytes of rabbit was about 156.25μg/mL.It was proved to be homogeneous by SDS-PAGE,authour measured subunit raletive molecular mass to be about 45 KDa.
The biochemical characteristics of IOL were studied.IOL contained 17.877% neutral saccharide.Its absorption peak of UV was at 269.3nm.In its amino acid composition,it had the highest content of Glu,followed by Asp,the lowest content was Met,but there was lack of Try.Agglutinating activities of IOL was only inhibited by barley-sugar.It was stable with activity at pH 4.0～9.0.It had high heat tolerance,and its peak temperature was 80℃.The addition of divalent metals(Zn~(2+)、Mg~(2+))influence the activity of the lectin.
The study on biologocal activity of IOL showed that it had different effects on 7 species of tested fungi(the growth of hyphal and spore germination)and 11 species of bacteria.IOL can agglutinate Scenedesmus obliquus,Dicrateria zhanjiangensis and Pavlova viridis and had effect on growth and content of chlorophyll-a of algae tested,it inhibited growth of Goniaulax polyramna and Phaeocystis globosa and reduced their content of chlorophyll-a,but promoted growth of Platymonas subcordiformis, Scenedesmus obliquus and Rhodella reticulata and increased content of chlorophyll-a.
理化特性的研究结果表明,铁钉菜凝集素含有17.877%的中性糖,最大紫外吸收峰为269.3nm;在氨基酸组成中,谷氨酸(Glu)的含量最高,天门冬氨酸(Asp)的含量次之,含量最低的是甲硫(蛋)氨酸(Met),不含色氨酸(Try)。凝集活性只被麦牙糖所抑制。在pH 4.0～9.0范围内相对稳定;具有较高的热稳定性,最高耐热温度为80℃;此外,IOL对二价金属离子Zn~(2+)和Mg~(2+)具有不同程度的依赖性。
生物学活性实验显示,IOL对7种供试真菌(菌丝生长及孢子萌发)及11种细菌生长具有不同程度的影响,能够凝集斜生栅藻、湛江叉鞭金藻和绿色巴夫藻,对供试藻类的生长及叶绿素a含量的影响不同,能抑制多纹膝沟藻和棕囊藻的生长,降低叶绿素a含量,但能够促进亚心型扁藻、斜生栅藻和红色蔷薇藻的生长,使它们叶绿素a的含量提高。
A lectin,named IOL,was isolated and purified from a brown marine alga Ishige okamurae by extraction with PBS,followed by ammonium sulfate precipitation and a combination of ion-exchange chromatography on DEAE-Sepharose fast flow and gel filtration on Sephadex G-100,and the lowest agglutinating concentration of IOL tested with erythrocytes of rabbit was about 156.25μg/mL.It was proved to be homogeneous by SDS-PAGE,authour measured subunit raletive molecular mass to be about 45 KDa.
The biochemical characteristics of IOL were studied.IOL contained 17.877% neutral saccharide.Its absorption peak of UV was at 269.3nm.In its amino acid composition,it had the highest content of Glu,followed by Asp,the lowest content was Met,but there was lack of Try.Agglutinating activities of IOL was only inhibited by barley-sugar.It was stable with activity at pH 4.0～9.0.It had high heat tolerance,and its peak temperature was 80℃.The addition of divalent metals(Zn~(2+)、Mg~(2+))influence the activity of the lectin.
The study on biologocal activity of IOL showed that it had different effects on 7 species of tested fungi(the growth of hyphal and spore germination)and 11 species of bacteria.IOL can agglutinate Scenedesmus obliquus,Dicrateria zhanjiangensis and Pavlova viridis and had effect on growth and content of chlorophyll-a of algae tested,it inhibited growth of Goniaulax polyramna and Phaeocystis globosa and reduced their content of chlorophyll-a,but promoted growth of Platymonas subcordiformis, Scenedesmus obliquus and Rhodella reticulata and increased content of chlorophyll-a.
引文
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