CAJ | 학술논문

从限氨固氮培养基中培养的缺失nif H的棕色固氮菌(Azotobacter vinelandii Lipmann)突变种DJ54中,分离纯化出部分纯的缺失FeMoco的钼铁蛋白(△nifH Av1).用相同纯化方法分别从DJ35和UW45突变种中纯化的△nifE Av1和NifB- Av1的纯度明显高于△nifH Av1的纯度.在合适的结晶条件下,可得到这三种蛋白的深棕色短斜四棱柱晶体.△nifH Av1与NifB- Av1一样,晶体形成所需的时间比△nifE Av1的长.而它结晶所需的沉淀剂和缓冲液最适浓度则与△nifE Av1的相同.SDS-PAGE鉴定表明,结晶的△nifH Av1与OP Av1的组成相似.这表明,在△nifH Av1溶液中形成的晶体可能就是该蛋白质的晶体.
종한안고담배양기중배양적결실nif H적종색고담균(Azotobacter vinelandii Lipmann)돌변충DJ54중,분리순화출부분순적결실FeMoco적목철단백(△nifH Av1).용상동순화방법분별종DJ35화UW45돌변충중순화적△nifE Av1화NifB- Av1적순도명현고우△nifH Av1적순도.재합괄적결정조건하,가득도저삼충단백적심종색단사사릉주정체.△nifH Av1여NifB- Av1일양,정체형성소수적시간비△nifE Av1적장.이타결정소수적침정제화완충액최괄농도칙여△nifE Av1적상동.SDS-PAGE감정표명,결정적△nifH Av1여OP Av1적조성상사.저표명,재△nifH Av1용액중형성적정체가능취시해단백질적정체.
A FeMoco-deficient △nifHAv1 was partially purified from a nifHdeleted mutant DJ54 of Azotobacter vinelandii Lipmann grown in NH3-limited medium. By using the same purification method, △nifE Av1 and NifB-Av1 were obtained from DJ35 and UW45, respectively. The latter two proteins were obviously much purer than △nifHAv1. Under a suitable condition for crystallization, dark brown short rhombohedron crystals could be obtained from the three proteins. Like NifB-Av1, the time for formation of △nifHAv1 crystal was longer than that of △nifEAv1. But the optimal concentrations of precipitant and buffer for crystallization of △nifH Av1 were similar to those of △nifE Av1. SDS-PAGE analysis showed that the crystalline △nifHAv1 was similar in the composition to OP Av1. It indicates that the crystal formed in △nifH Av1 solution could be the protein crystal.