| 小麦纹枯病菌胞外蛋白酶的纯化与特性 |
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| 引用本文： | 赵蕾,张天宇.小麦纹枯病菌胞外蛋白酶的纯化与特性[J].植物病理学报,2006,36(2):135-141. |
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| 作者姓名： | 赵蕾 张天宇 |
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| 作者单位： | 1. 山东农业大学植物保护学院, 泰安 271018;2. 山东师范大学生命科学学院, 济南 250014 |
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| 摘 要： | 小麦纹枯病菌(Rhizoctonia cerealis Vander Hoeven)在以小麦细胞壁为唯一碳氮源的液体培养基中可产生多种蛋白酶,培养液经硫酸铵沉淀,SP-Sepharose Fast Flow离子交换层析,Phenyl(high-sub)-Sepharose Fast Flow疏水层析。Sephadex G-75分子筛层析,分离纯化到分子量约40kD的一种蛋白酶。该酶能水解胰蛋白酶专一性底物Benz-Phe-Val-Arg-NA,对凝乳弹性蛋白酶底物Sue-Ala-Ala-Pro-Phe-NA和枯草杆菌蛋白酶底物CBZ-Gly-Gly-Leu-NA无活性,能被胰蛋白酶抑制剂aprotinin、leupeptin和soybean trypsin inhibitor强烈抑制,丝氨酸蛋白酶抑制剂PMSF能部分抑制酶的活性。表明该蛋白酶是丝氨酸蛋白酶中的一种类胰蛋白酶。
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| 关 键 词： | 小麦纹枯病菌 胞外蛋白酶 类胰蛋白酶 |
| 文章编号： | 0412-0914(2006)02-0135-07 |
| 修稿时间： | 2005年3月10日 |
Purification and characterization of an extracellular protease from Rhizoctonia cerealis |
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| ZHAO Lei,ZHANG Tian-yu.Purification and characterization of an extracellular protease from Rhizoctonia cerealis[J].Acta Phytopathologica Sinica,2006,36(2):135-141. |
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| Authors: | ZHAO Lei ZHANG Tian-yu |
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| Institution: | 1. College of Plant Protection, Shandong Agricultural University, Taian 271018, China;2. College of Life Science, Shandong Normal University, Jinan 250014, China |
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| Abstract: | Rhizoctonia cerealis could secrete several extracellular proteases when grown in liquid medium with wheat cell walls as the sole carbon and nitrogen source.A kind of protease was purified after ammonium sulfate precipitation,SP-Sepharose Fast Flow chromatography,Phenyl high-sub-Sepharose Fast Flow chromatography and Sephadex G-75 chromatography.It appeared as a single band corresponding to molecular weight MWof approximately 40 kD on SDS-PAGE with silver staining.It showed high activities against the trypsin substrate Benz-Phe-Val-Arg-NA,and also the substrates D-Val-Leu-Arg-NA,Benz-Pro-Phe-Arg-NA and D-Val-Phe-Lys-NA.It was strongly inhibited by aprotinin,leupeptin,soybean trypsin inhibitor and partly inhibited by PMSF,indicating that it is quite probably a trypsin-like protease. |
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| Keywords: | Rhizoctonia cerealis extracellular protease trypsin-like protease |
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