Composite

Part:BBa_K4275037

Designed by: Zihuan Zhang   Group: iGEM22_GreatBay_SCIE   (2022-10-11)
Revision as of 10:20, 12 October 2022 by Jerryatcg (Talk | contribs) (→‎Sequence and Features)


pLAC4-KmarxMFα-TaLPMO-t-tTDH1

The composite part includes K. marx mating factor alpha (BBa_K4275000) and cellulase booster TaLPMO-t fused with type I dokerin for the assembly of cellulosome complex.

The fusion of K. marx mating factor alpha is used in order for the target TaLPMO-t cellulase booster to be secreted from our host yeast, which would be anchored on scaffold protein and assembled into the cellulosome complex.


Usage and Biology

Kluyveromyces marxianus mating factor alpha act as a secretion signal in Kluyveromyces marxianus. The mating factor alpha fuses an alpha mating factor domain onto TaLPMO-t. The signal peptide on the domain would direct the TaLPMO-t into RER and Golgi body, enabling the designated cellulase booster to be secreted by Kluyveromyces marxianus.

TaLPMO-t is a cellulase booster with type I dokerin fused on its C terminal to convert the protein from free booster enzyme into cellulosomal mode. As a copper-dependent enzyme, TaLPMO-t oxidizes and cleaves glycosidic bonds in cellulose, which significantly boosts the efficiency of crystalline cellulose degradation.


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 XhoI site found at 1252
    Illegal XhoI site found at 2416
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 331
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 331
    Illegal AgeI site found at 1554
    Illegal AgeI site found at 1903
  • 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. Harris, Paul V. et al. "Stimulation Of Lignocellulosic Biomass Hydrolysis By Proteins Of Glycoside Hydrolase Family 61: Structure And Function Of A Large, Enigmatic Family". Biochemistry, vol 49, no. 15, 2010, pp. 3305-3316. American Chemical Society (ACS), https://doi.org/10.1021/bi100009p.
4. "Part:Bba K2753052 - Parts.Igem.Org". Parts.Igem.Org, 2022, https://parts.igem.org/Part:BBa_K2753052.


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