J Integr Plant Biol. ›› 2003, Vol. 45 ›› Issue (4): 427-431.

• Research Articles • Previous Articles     Next Articles

Crystallization of Nitrogenase MoFe Protein from a Mutant nifE Deleted Strain of Azotobacter vinelandii

ZHAO Ying, ZHAO Jian-Feng, Lü Yu-Bing, WANG Zhi-Ping WANG Yao-Ping, HUANG Ju-Fu   


Under a suitable condition of crystallization, dark brown short rhombohedron crystals could be obtained from FeMoco-deficient MoFe protein (ΔnifE Av1) purified from a nifE deleted mutant DJ35 of Azotobacter vinelandii Lipmann grown in NH3-limited medium. The number, size and quality of crystals were significantly affected by either the concentration of precipitants and buffer or diffusion method. The longest sides of the largest crystal of ΔnifE Av1 protein, which was obtained by vapor diffusion in the hanging drop method, were 0.12 and 0.13 mm, respectively. Key words: mutant DJ35 of Azotobacter vinelandii; nitrogenase ΔnifE Av1; crystallization

棕色固氮菌缺失nifE 的突变种固氮酶钼铁蛋白的结晶
赵 颖  赵剑峰  吕玉兵  汪志平  王耀萍  黄巨富

(1. 中国科学院植物研究所光合作用与环境分子生理学重点实验室,北京100093;2.  中国科学院生物物理研究所,北京100101; 3.  浙江大学原子核农业研究所,杭州310029)

摘要: 从限氨固氮培养基中培养棕色固氮菌(Azotobacter vinelandii Lipmann)缺失nifE的突变种DJ35中,分离纯化得到缺失FeMoco的钼铁蛋白(Δnif E Av1).在一定条件下结晶得到深棕色短斜四棱柱晶体.结晶溶液中各组分的浓度以及结晶方法等对其晶核数目、晶体大小和质量有明显影响.目前用气相扩散的悬滴法所得的最大晶体的二维边长分别为0.12 mm和0.13 mm.

关键词: 棕色固氮菌DJ35突变种;固氮酶Δnif E 钼铁蛋白;结晶
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