Designed by: Shirin Bamezai   Group: iGEM22_Imperial_College_London   (2022-09-29)


This part codes for a fusion protein consisting of the B. subtilis native CotG anchor protein and a B. pumilus chitinase enzyme. The CotG protein has a midspore localization and high abundance in B. subtilis spores, enabling spore display. In this case with the chitinase fusion, spores will display chitinases on the surface capable of breaking down chitin into its monomer components, such as N-acetylglucosamine. In the context of our project, the chitinase activity enables us to equip spores with the ability to breakdown the chitin-rich outer coat of fungal pathogens, not only inhibiting their activity but generating chitin monomers that can function as a biomarker, as well as an inducer of plant systemic immune response. This fusion includes a flexible linker, to reduce steric hindrance between the two protein domains and enable greater chitinase activity. The same was selected for testing following dry lab modelling.

Usage and Biology

It was shown by previous literature that the CotG coat protein of B.Subtilis can be used to display other proteins, which provide additional functions. Chitinase is an enzyme, which can break down chitin into N-acetyl-glucosamine (NAG): being able to break down chitin polymers into individual monomers and being able to sense them to indicate the presence of a fungal pathogen is a crucial preliminary aspect of our fungicidal response.

Figure 1. The schematic design of CotG-3Gx3-Chis


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[3] Hamid R, Khan MA, Ahmad M, Ahmad MM,Abdin MZ, Musarrat J, Javed S. Chitinases: An update. J Pharm Bioallied Sci.2013 Jan;5(1):21-9. doi: 10.4103/0975-7406.106559. PMID: 23559820; PMCID:PMC3612335.

[4] ina Adrangi, Mohammad Ali Faramarzi,From bacteria to human: Ajourney into the world of chitinases, Biotechnology Advances, Volume 31, Issue8, 2013, Pages 1786-1795, ISSN 0734-9750,

[5]McKenney, P., Driks, A. & Eichenberger, P. The Bacillus subtilis endospore:assembly and functions of the multilayered coat. Nat Rev Microbiol 11,33–44 (2013).

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Sequence and Features

Assembly Compatibility:
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    Illegal EcoRI site found at 885
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    Illegal EcoRI site found at 885
  • 21
    Illegal EcoRI site found at 885
    Illegal BamHI site found at 870
  • 23
    Illegal EcoRI site found at 885
  • 25
    Illegal EcoRI site found at 885
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