Composite

Part:BBa_K4275039

Designed by: Zihuan Zhang   Group: iGEM22_GreatBay_SCIE   (2022-10-11)
Revision as of 10:22, 12 October 2022 by Jerryatcg (Talk | contribs) (→‎Sequence and Features)


pLAC4-KmarxMFα-MtCDH-t-tTDH1

The composite part is constructed from Kluyveromyces marxianus mating factor alpha (BBa_K4275000) and cellulase booster MtCDH-t fused with type I dockerin (BBa_K4275005).

The fusion of Kluyveromyces marxianus mating factor alpha (BBa_K4275000) ensures effective secretion of MtCDH-t protein by the host yeast, thus avoiding lysing the yeast cell to extract the designated cellulase booster.

Usage and Biology

Kluyveromyces marxianus mating factor alpha is an secretion signal in Kluyveromyces marxianus, which encodes for an alpha mating factor domain fused with target protein MtCDH-t. The signal peptide in alpha mating factor domain would guide the protein into RER and Golgi apparatus for the secretion of MtCDH-t.

MtCDH-t is considered a cellulase booster with strong reductive nature, enabling it to act as an electron donor to another cellulase booster TaLPMO-t during crystalline cellulose degradation. The free cellulase booster is converted into cellulosomal mode via the fusion of a type I dockerin to its C terminal. Functioning synergistically with four other cellulase and booster enzymes, MtCDH-t contributes towards effective biodegradation of cellulose components.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 331
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 331
    Illegal NheI site found at 89
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 331
    Illegal BglII site found at 3987
    Illegal XhoI site found at 1252
    Illegal XhoI site found at 4150
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 331
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 331
    Illegal NgoMIV site found at 2788
    Illegal AgeI site found at 1639
    Illegal AgeI site found at 2026
    Illegal AgeI site found at 2152
    Illegal AgeI site found at 2701
  • 1000
    COMPATIBLE WITH RFC[1000]


References

1. Rajkumar, Arun S. et al. "Biological Parts For Kluyveromyces Marxianus Synthetic Biology". Frontiers In Bioengineering And Biotechnology, vol 7, 2019. Frontiers Media SA, https://doi.org/10.3389/fbioe.2019.00097.
2.“Mating Factor Alpha-1 [Kluyveromyces Marxianus] - Protein - NCBI.” National Center for Biotechnology Information, U.S. National Library of Medicine, https://www.ncbi.nlm.nih.gov/protein/QGN17207.1
3. Phillips, Christopher M. et al. "Cellobiose Dehydrogenase And A Copper-Dependent Polysaccharide Monooxygenase Potentiate Cellulose Degradation By Neurospora Crassa". ACS Chemical Biology, vol 6, no. 12, 2011, pp. 1399-1406. American Chemical Society (ACS), https://doi.org/10.1021/cb200351y.
4. "Part:Bba K2753052 - Parts.Igem.Org". Parts.Igem.Org, 2022, https://parts.igem.org/Part:BBa_K2753052.


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