Coding

Part:BBa_K4607000

Designed by: Axel Rojero   Group: iGEM23_Tec-Chihuahua   (2023-07-20)
Revision as of 19:08, 25 July 2023 by Axelrohz24 (Talk | contribs) (→‎Usage and Biology)


LysCSA13-ABD

lyscsa13-cds.jpg
Figure 1. LysCSA13-ABD protein diagram.

This biobrick is a fusion protein based on the endolysin (Lys) from Staphylococcus aureus virulent bacteriophage CSA13 [1], and the albumin binding domain (ABD) from streptococcal protein G [2]. Its purpose is to recognize the S. aureus cell wall, and degrade it. The part is adapted to the Golden Gate cloning method. This part also contains a TEV cleavage site for the removal of the x6HisTag after the protein purification process.

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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Usage and Biology

The endolysin Lys from the Staphylococcus aureus virulent bacteriophage CSA1, is composed of two domains. The bacteriophage CSA13 CHAP domain has excellent catalytic activity, up to 90%, degrading almost 15 strains of Staphylococcus including methicillin-resistant strains. As with many of the endolysins, it cleaves to the cell wall by disrupting the peptidoglycan that composes the bacterial cell; for this to be possible, the bacteriophage CSA13 SH3 domain recognizes and binds to the glycine of the pentaglycine cross-bridge glycosidic bond in the heteropolymer of the peptidoglycan, which makes it completely safe for the host [1].

The use of enzybiotics represents an alternative to the misuse of antibiotics without loss of efficiency; it is a novel and environmentally friendly process. It supplies antibacterial protection to pathogenic bacteria but shows no toxic effects on mammalian cells. Our protein has an extra region, the albumin binding domain, that causes an important increase in the life-time of the fusion protein [2].

lyscsa13-protein-param.jpg
Table 1. LysCSA13-ABD protein parameters.

Results

lyscsa13-cds2.jpg
Figure 2. 3D structure of the LysCSA13-ABD protein, obtained with AlphaFold2.

References

[1] Cha, Y., Son, B., & Ryu, S. (2019). Effective removal of staphylococcal biofilms on various food contact surfaces by Staphylococcus aureus phage endolysin LysCSA13. Food Microbiology, 84, 103245. https://doi.org/10.1016/j.fm.2019.103245

[2] Seijsing, J., Sobieraj, A. M., Keller, N., Shen, Y., Zinkernagel, A. S., Loessner, M. J., & Schmelcher, M. (2018). Improved Biodistribution and Extended Serum Half-Life of a Bacteriophage Endolysin by Albumin Binding Domain Fusion. Frontiers in Microbiology, 9. https://doi.org/10.3389/fmicb.2018.029
[edit]
Categories
Parameters
None