CAJ | 학술논문

运用传统的层析技术对膜荚黄芪种子中两种几丁质酶进行分离纯化,并对分离纯化中各个步骤得到的蛋白进行了活性研究.结果表明:(1)粗提液的硫酸铵沉淀经再生几丁质亲和柱和凝胶过滤层析Sephadex G-75得到几丁质酶A.(2)粗提液的硫酸铵沉淀经离子交换色谱DE-52、CM、凝胶过滤层析Sephadex G-75得到几丁质酶B.(3)几丁质酶A、B的比活性分别为35.6 U/mg和4.1 U/mg.(4)从膜荚黄芪种子中分离纯化的几丁质酶A和B均为糖蛋白,含糖量分别为6.1%和5.8%.(5) SDS-PAGE显示,几丁质酶A、B的分子量分别为35.5 ku、39.6 ku;凝胶过滤层析测定几丁质酶A、B的分子量分别为36.9 ku、40.8 ku;表明几丁质酶A、B均为单亚基蛋白.
운용전통적층석기술대막협황기충자중량충궤정질매진행분리순화,병대분리순화중각개보취득도적단백진행료활성연구.결과표명:(1)조제액적류산안침정경재생궤정질친화주화응효과려층석Sephadex G-75득도궤정질매A.(2)조제액적류산안침정경리자교환색보DE-52、CM、응효과려층석Sephadex G-75득도궤정질매B.(3)궤정질매A、B적비활성분별위35.6 U/mg화4.1 U/mg.(4)종막협황기충자중분리순화적궤정질매A화B균위당단백,함당량분별위6.1%화5.8%.(5) SDS-PAGE현시,궤정질매A、B적분자량분별위35.5 ku、39.6 ku;응효과려층석측정궤정질매A、B적분자량분별위36.9 ku、40.8 ku;표명궤정질매A、B균위단아기단백.
Two chitinases were purified from the seeds of Astragalus membranaceus (Fisch.) Bge.using traditional chromatographies and the activities of the proteins obtained during each purification step was determined.Results showed:(1)a chitinase (designated as chitinase A) was purified using a combination of 20-60 % ammonium sulfate saturation,regenerated chitin column and Sephadex G-75.(2)Another chitinase (designated as chitinase B) was purified by a procedure containing ammonium sulfate precipitation,ion-exchange chromatographies on DE-52 and CM,gel filtration on Sephadex G-75.(3)The specific activities of chitinase A and chitinase B were 35.6 U/mg and 4.1 U/mg,respectively.(4)Both chitinase A and chitinase B were glycoproteins with a neutral carbohydrate content of 6.1% and 5.8%,respectively.(5)Both chitinase A and chitinase B appeared as a single band with a molecular mass of 35.5 and 39.6 ku on SDS-PAGE and the relative molecular masses were 36.9 and 40.8 ku estimated by gel filtration on a calibrated Sephacryl S-200 column,indicating that both of the two chitinases are monomeric proteins.