Part:BBa_K2135001

petF

petF codes for ferredoxin, an important electron donor in Synechocystis that is mainly works to be the final electron acceptor in the electron transport chain during photosynthesis.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Usage and Biology

petF codes for ferredoxin, an important electron donor in Synechocystis that is the final electron acceptor in the electron transport chain during photosynthesis[1]. Being the final electron acceptor, it has the greatest reducing power. Ferredoxin is also the reducing agent in many other cellular reactions. Notably, in diazotrophs (cells that fix nitrogen), ferredoxin is a co-factor for nitrogenase activity, allowing the bacteria to fix nitrogen gas and convert it ammonia for the cell to use. Some important uses of ferredoxin (Fd) are illustrated in the picture below:

Source:http://www.nature.com/nsmb/journal/v8/n2/index.html

Other Uses

Overexpression of petF results in the overproduction of ferredoxin in the cell. The resulting cells have a distinct red color (picture below), most likely due to increased cytochrome activity[2]. The extra ferredoxin can be either reduced or oxidized and can play a part in various cellular activates. For example, the [http://2016.igem.org/Team:WashU_StLouis WashU_STL 2016 team] used petF in conjunction with an inducible TetR-pTet promoter to create a composite part that can control the levels of ferredoxin produced. See the composite part page for data and uses.

The top test tube shows the distinct red color of the cells with overproduction of ferredoxin while the bottom test tube shows the control group.

Data

Below is data for biotin production (using a biotin assay) when petF is combined with an inducible promoter. Biotin is produced in an electron-dependent biosynthesis, so the amount of biotin produced is correlated to the amount of usable reduced ferredoxin in the cell[3].

References

1. BLASCHKOWSKI, H. P., KNAPPE, J., LUDWIG-FESTL, M. and NEUER, G. (1982), Routes of Flavodoxin and Ferredoxin Reduction in Escherichia coli. European Journal of Biochemistry, 123: 563–569. doi:10.1111/j.1432-1033.1982.tb06569.x
2. Haddock, B. A., J. ALLAN Downie, and P. B. Garland. "Kinetic characterization of the membrane-bound cytochromes of Escherichia coli grown under a variety of conditions by using a stopped-flow dual-wavelength spectrophotometer." Biochemical Journal 154.2 (1976): 285-294.
3. Lin, Steven, and John E. Cronan. "Closing in on complete pathways of biotin biosynthesis." Molecular BioSystems 7.6 (2011): 1811-1821.