| Autor: |
Tatemichi, Yuki, Nakahara, Takeharu, Ueda, Mitsuyoshi, Kuroda, Kouichi |
| Zdroj: |
Bioscience, Biotechnology, and Biochemistry; September 2021, Vol. 85 Issue: 10 p2209-2216, 8p |
| Abstrakt: |
Biological nitrogen fixation by nitrogenase has attracted attention as an alternative method to chemical nitrogen fixation, which requires large amounts of fossil fuels. Azotobacter vinelandii, which produces an oxygen-sensitive nitrogenase, can fix nitrogen even under aerobic conditions; therefore, the heterologous expression of nif-related genes from A. vinelandiiis a promising strategy for developing a biological nitrogen fixation method. We assembled 17 nif-related genes, which are scattered throughout the genome of A. vinelandii, into synthetic gene clusters by overlap-extension-PCR and seamless cloning and expressed them in Escherichia coli. The transcription and translation of the 17 nif-related genes were evaluated by RT-qPCR and LC-MS/MS, respectively. The constructed E. colishowed nitrogenase activity under anaerobic and microaerobic conditions. This strain would be a useful model for examining the effect of other genes from A. vinelandiion nitrogen fixation by expressing them in addition to the minimal set of nif-related genes.Graphical AbstractAssembly of 17 nif-related genes into plasmids and expression in E. coli. |
| Databáze: |
Supplemental Index |
| Externí odkaz: |
|
