LARGE-SCALE PURIFICATION OF HIGH-ACTIVITY AZOTOBACTER-VINELANDII NITROGENASE
LARGE-SCALE PURIFICATION OF HIGH-ACTIVITY AZOTOBACTER-VINELANDII NITROGENASE
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DOI：
10.1016/0005-2744(80)90180-1
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发表时间：
1980-01-01
期刊：
影响因子：
--
通讯作者：
STIEFEL, EI
中科院分区：
文献类型：
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作者：
BURGESS, BK;JACOBS, DB;STIEFEL, EI
A large scale, rapid, high-yield purification procedure for A. vinelandii nitrogenase proteins was developed. Yields of approximately 600 mg of the FeMO protein (Av1) and approximately 550 mg of the Fe protein Av2 are routinely obtained using a procedure that requires only 28 h. The specific activities of Av1 and Av2, respectively, are 3000 and 2100 nmol H2 evolved/min per mg. These activities are significantly higher than those commonly used in reactivity studies. Procedures for the isolation and concentration of large quantities of Fe-Mo cofactor of nitrogenase are also reported. Techniques for anaerobic protein manipulation, generally applicable to the purification of O2 sensitive proteins are also described.
