Part:BBa_K3841010

Particulate methane monooxygenase alpha subunit (PmoB)

Methane monooxygenases catalyses the initial oxygenation of methane to methanol in methanotrophic bacteria [1].

The bacterium Methylococcus capsulatus has two forms of methane monooxygenase, a soluble (sMMO) and a membrane-bound (particulate) type (pMMO). The particulate type is a cupper-dependent [2] nonamer composed of three alpha:beta:gamma heterotrimeric protomers assembled into a cylindrical structure; the beta and gamma subunits comprise the bulk of the membrane-spanning regions and the soluble regions are derived primarily from alpha subunits which form two antiparallel beta-barrel-like structures each [3].

BBa_K3841010 is the alpha subunit of pMMO and has been codon-optimized for the methylotropich yeast, Komagataella phaffii.

Folding of PmoB (BBa_K3841010) with signal peptide (BBa_K3841036) and PmoB (BBa_K3841010) simulated using AlphaFold [4]. This part follows the pink structure.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 619
  Illegal XbaI site found at 1153
  
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 334
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 619
  Illegal XbaI site found at 1153
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 619
  Illegal XbaI site found at 1153
  
• 1000
  COMPATIBLE WITH RFC[1000]

References

[1] Hanson, Richard S., and Thomas E. Hanson. “Methanotrophic Bacteria.” Microbiological Reviews, vol. 60, no. 2, American Society for Microbiology, 1996, pp. 439–71, doi:10.1128/mmbr.60.2.439-471.1996.

[2] Basu, Piku, et al. “The Membrane-Associated Form of Methane Mono-Oxygenase from Methylococcus Capsulatus (Bath) Is a Copper/Iron Protein.” Biochemical Journal, vol. 369, no. 2, PORTLAND PRESS LTD, 2003, pp. 417–27, doi:10.1042/BJ20020823.

[3] Lieberman, Raquel L., and Amy C. Rosenzweig. “Crystal Structure of a Membrane-Bound Metalloenzyme That Catalyses the Biological Oxidation of Methane.” Nature, vol. 434, no. 7030, NATURE PUBLISHING GROUP, 2005, pp. 177–82, doi:10.1038/nature03311.

[4] Cramer, Patrick. “AlphaFold2 And the Future of Structural Biology.” Nature Structural and Molecular Biology, vol. 28, no. 9, Nature Research, 2021, pp. 704–05, doi:10.1038/s41594-021-00650-1.