Part:BBa_K3692003

SED1

Usage and Biology

Sed1 is important in organizing the yeast cell wall (Hossain et al., 2020).

As the yeast cell wall provides space to display heterologous proteins, they can be anchored to it by covalent linking with GPI (glycosylphosphatidylinositol)-anchored cell wall proteins (GPI-CWP) domain (Inokuma et al., 2020). GPI-anchored proteins have been identified as adhesion molecules, activation antigens and other miscellaneous glycoproteins (Stewart & Stewart, 2017).

Anchorage position of the target protein is dependent on the cell wall protein location. One of the well-known GPI-anchoring domains is Sed1.

One study showed that eGFP fused with the Sed1-anchoring domain was localized to the external surface of the cell wall (Inokuma et al., 2020).

It has been shown that the anchoring domain from different GPI cell wall proteins exhibit different efficiencies for the cell-surface display of target enzymes (Andreu & del Olmo, 2018).

The cell of Sacharomyces cerevisiae consists of β-glucans and mannoproteins where the hydrolysis of former ones is catalyzed by ß-glucanases (Orlean, 2012).

We have shown previously that induced expression of bacterial glucanases has the potential to cause lysis in S. cerevisiae (Team:Tartu TUIT - 2019.Igem.Org, n.d.). The efficiency of autolysis could depend on the localization of glucanases, which can be anchored to the yeast cell wall by fusing with GPI-CWPs (Inokuma et al., 2020)

References

• Hossain, S. A., Rahman, S. R., Ahmed, T., & Mandal, C. (2020). An overview of yeast cell wall proteins and their contribution in yeast display system. Asian Journal of Medical and Biological Research, 5(4), 246–257. https://doi.org/10.3329/ajmbr.v5i4.45261
• Inokuma, K., Kurono, H., den Haan, R., van Zyl, W. H., Hasunuma, T., & Kondo, A. (2020). Novel strategy for anchorage position control of GPI-attached proteins in the yeast cell wall using different GPI-anchoring domains. Metabolic Engineering, 57, 110–117. https://doi.org/10.1016/j.ymben.2019.11.004
• Stewart, G. G., & Stewart, G. G. (2017). History of Brewing and Distilling Yeast. In Brewing and Distilling Yeasts (pp. 11–36). Springer International Publishing. https://doi.org/10.1007/978-3-319-69126-8_2
• Andreu, C., & del Olmo, M. (2018). Yeast arming systems: pros and cons of different protein anchors and other elements required for display. In Applied Microbiology and Biotechnology (Vol. 102, Issue 6, pp. 2543–2561). Springer Verlag. https://doi.org/10.1007/s00253-018-8827-6
• Orlean, P. (2012). Architecture and biosynthesis of the Saccharomyces cerevisiae cell wall. In Genetics (Vol. 192, Issue 3, pp. 775–818). https://doi.org/10.1534/genetics.112.144485
• Team:Tartu TUIT - 2019.igem.org. (n.d.). Retrieved October 16, 2020, from https://2019.igem.org/Team:Tartu_TUIT

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 391
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 328