We previously constructed an Escherichia coli strain expressing 16 nitrogen fixation (nif) and 2 nif-related genes from Azotobacter vinelandii and improved nitrogenase activity to some extent by enhancing NifH-related functions. In the present study, we analyzed the formation of dinitrogenase, a heterotetrameric NifDK, produced in E. coli, using gel-filtration chromatography and blue native PAGE to gain insight into further increases in nitrogenase activity. A certain proportion of NifD and NifK proteins produced in E. coli were present as the complete NifDK component, but some remained in the intermediate stages of maturation. Overexpression of nafY, which is involved in holo-NifDK formation, effectively increased nitrogenase activity.

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http://dx.doi.org/10.2323/jgam.2024.12.001DOI Listing

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View Article and Find Full Text PDF

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