Coding

Part:BBa_K3841010

Designed by: Emil Ørsted Christensen   Group: iGEM21_DTU-Denmark   (2021-09-02)


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.


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