Work

Characterization of the Azotobacter vinelandii nitrogenase complex expressed in Escherichia coli toward further activity improvement

Year: 2024

Type: article

Abstract: 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 NifD2K2, 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 NifD2K2 component, but some remained in the intermediate stages of maturation. Overexpression of nafY, which is involved in holo-NifD2K2 formation, effectively increased nitrogenase activity.

Source: The Journal of General and Applied Microbiology

DOI: 10.2323/jgam.2024.12.001

Authors: Yusuke Ito, Daisuke Yoshidome, Makoto Hidaka, Yasuko Araki, Kotaro Ito, Saori Kosono, Makoto Nishiyama

Institutions: Kikkoman (Japan), The University of Tokyo, Collaborative Research Institute for Innovative Microbiology, The University of Tokyo

Language: English

Topic: Metalloenzymes and iron-sulfur proteins

Subfield: Biochemistry

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: diamond

OpenAlex ID: W4406203327

FWCI:

Cites: 21

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Related to: 10