| 纪兆林，郭 菊，张丽娜，陈夕军，童蕴慧，徐敬友.小麦纹枯病菌多聚半乳糖醛酸酶及其理化性质研究[J].麦类作物学报,2012,32(3):569 |
| 小麦纹枯病菌多聚半乳糖醛酸酶及其理化性质研究 |
| Polygalacturonase from Rhizoctonia cerealis, the Pathogen of Wheat Sharp Eyespot and Characterization of Its Physiological and Biochemistry Properties |
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| DOI：10.7606/j.issn.1009-1041.2012.03.034 |
| 中文关键词: 小麦纹枯病菌 多聚半乳糖醛酸酶 理化性质 |
| 英文关键词:Rhizoctonia cerealis Polygalacturonase Characteristics |
| 基金项目:国家转基因重大专项（2009ZX08001 014B）；江苏省自然科学基金项目（BK2010305）；扬州大学高层次人才科研启动基金项目（0274983014811）。 |
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| 中文摘要: |
| 为了解小麦纹枯病菌产生的多聚半乳糖醛酸酶（Polygalacturonase, PG）的理化性质，先用丙酮法提取PG粗蛋白，再经DEAE Sepharose Fast Flow离子交换柱、Phenyl Sepharose 6 Fast Flow疏水柱和Sephadex G 75凝胶柱纯化，得到一种具有较高活性的PG纯蛋白。该蛋白分子量为41.78 kD；等电点为5.34；含有糖基，含糖量为9.10%；含有α氨基酸，但不含芳香族氨基酸；不含脂基。这种蛋白在pH 4~12范围内均具有活性，pH为6时活性最大；对热不稳定，100 ℃下水浴20 min，活性完全丧失；对胰蛋白酶和蛋白酶K敏感，酶处理后其活性只有对照的15.7%和10.2%；对紫外线和氯仿亦敏感，处理后活性仅为对照的18.6%和14.9%。 |
| 英文摘要: |
| Polygalacturonase (PG) produced from Rhizoctonia cerealis, the pathogen of wheat sharp eyespot, was one of the most important pathogenic factors. The crude protein of PG in culture filtrate was precipitated with acetone and then purified by DEAE Sepharose Fast Flow ion exchange column, Phenyl Sepharose 6 Fast Flow hydrophobic column and Sephadex G 75 gel column in turn. Molecular weight of the purified PG was 41.78 kD by SDS PAGE, and pI value was 5.34 by IEF PAGE. The protein was demonstrated as glycoprotein with 9.10% saccharide content, and which also contained α amino acid, but no proline or hydroxyproline and lipid. The activity of the protein was observed in the range of pH 4 to 12, with strongest activity at pH 6. The protein lost activity at 100 ℃ after 20 min, and it was also sensitive to trypsin, proteinase K, ultraviolet radiation and chloroform. After being treated with above methods, the activity of PG was only 15.7%, 10.2%, 18.6% and 14.9% of the contrast, respectively. |
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