嗜酸乳杆菌3-磷酸甘油醛脱氢酶的分离纯化  

Purification and characterization of glyceraldehyde-3-phosphate dehydrogenase from Lactobacillus acidophilus

作　　者：潘渠[1,2] 丛延广[1] 侯瑞[1] 陈志谨[1] 胡福泉[1] 邱岳[2] 王广科[2] 

机构地区：[1]第三军医大学基础部微生物学教研室 [2]成都医学院病原教研室

出　　处：《免疫学杂志》2009年第4期461-464,共4页Immunological Journal

基　　金：四川省科技攻关项目(05SG022-019)资助

摘　　要：目的分离纯化嗜酸乳杆菌(Lactobacillus acidophilus)ATCC 4356的3-磷酸甘油醛脱氢酶。方法利用硫酸铵分级沉淀法、亲和层析、阴离子交换和分子筛层析对嗜酸乳杆菌GAPDH进行了分离纯化。通过N端测序鉴定了纯化产物;并用分子筛测定了嗜酸乳杆菌GAPDH的相对分子质量(Mr)。结果纯化产物在SDS-PAGE胶上呈现Mr为3.7×104的单一条带,产量为0.9 mg/L(嗜酸乳杆菌培养物)。N端测序结果证明纯化产物为嗜酸乳杆菌GAPDH。分子筛测定的GAPDH的Mr为7.08×104,表明嗜酸乳杆菌GAPDH是二聚体。结论通过该纯化方案,可大量获得电泳纯的嗜酸乳杆菌GAPDH,为进一步研究其免疫功能以及开发为免疫促进剂打下了基础。Objective To purify and characterize glyceraldehyde-3-phosphate dehydrogenase(GADPH) from L.acidophilus ATCC 4 356.Methods GADPH was purified through procedures of ammonium sulphate fractionation,affinity chromatography,anion-exchange,and sieve chromatography.Then the purified product was characterized by N-terminal sequencing.The apparent molecular weight of homogenous GADPH was measured by molecular sieve.Results The purified product showed a single band of 3.7×104 and was identified as GADPH of L.acidoph...

关 键 词：3-磷酸甘油醛脱氢酶 嗜酸乳杆菌 纯化 N端测序 二聚体 

分 类 号：R914[医药卫生—药物化学]