论文部分内容阅读
本文采用硫酸铵沉淀法,分别以生理盐水,蒸馏水,pH=7.2的Tris-HCl缓冲液,pH=7.2的磷酸缓冲液为浸提液,对西兰苔凝集素进行提取,然后依次利用DEAE-52阴离子交换层析和SephadexG-75对凝集素粗品进行分离纯化,并对纯化后的西兰苔凝集素的部分性质进行研究。结果表明:pH=7.2的Tris-HCl缓冲液的提取率最高,为3.57±0.02%;纯化后的西兰苔凝集素分子量约为27 ku,糖含量为7.60±0.03%,其凝血活力为28;糖抑制试验表明,西兰苔凝集素的凝集活性受D-甘露糖和阿拉伯糖的抑制;热稳定性实验表明90℃以下凝集素的凝集活性不受温度影响,有较好的热稳定性。此外,西兰苔凝集素的最适pH范围是7~8,强酸强碱环境中其凝血活性明显下降。在含有Ca2+的溶液中其凝集活性有所增强,表明西兰苔凝集素活性受Ca2+影响较大。
In this paper, ammonium sulfate precipitation method, respectively, saline, distilled water, pH = 7.2 Tris-HCl buffer, pH = 7.2 phosphate buffer solution for extraction of zeaxanthin agglutinin, and then followed by DEAE- 52 anion exchange chromatography and SephadexG-75 on the purification of crude lectin, and purified part of the properties of zealand lectin. The results showed that the highest extraction rate was 3.57 ± 0.02% for Tris-HCl buffer solution with pH = 7.2. The molecular weight of purified zeaxanthin was about 27 ku, the content of sugar was 7.60 ± 0.03% and the coagulation activity was 28 ; Sugar inhibition test showed that the agglutination activity of zeaxan Lectin was inhibited by D-mannose and arabinose; thermal stability experiments showed that agglutination activity at 90 ℃ was not affected by temperature, and had good thermal stability . In addition, the optimum pH range for zeaxanthin lectin is 7-8, and the coagulation activity is significantly decreased in strong acid and alkali environment. Aggregation activity was enhanced in the Ca2 + -containing solution, indicating that the activity of Lectin was significantly affected by Ca2 +.