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     <td align="center"><img src="https://static.igem.wiki/teams/4897/wiki/parts/turboid-fgb-staining.png" width="800" height="auto" /> </td>
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     <td align="center">Fig. 2. Amplification results of P. acne bacterial DNA by BS DNA-322 under the competition of randomly generated DNA fragment of size 322.</td>
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     <td align="center">Fig. 1. Staining Result of P. acne by Streptavidin-568 Binding to Activated Biotin through Oil Immersion. </td>
 
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Latest revision as of 13:36, 11 October 2023


GPX7, Caf1-AMP, and TurboID-FGB for Eliminating P. acne

What is it?

Here we use GPX7 expression system(BBa_K4897006) and Caf1-AMP(BBa_K4897007) assembly system to generate the key component of BS cleanser. Also, TurboID-FGB is also added to this composite part to eliminate P. acne.

Glutathione peroxidase7 (GPX 7) is an antioxidant enzyme in mammals in response to oxidative stress [1]. Oxidative stress essentially causes the aging of the skin and is typically caused by reactive oxygen species [2]. One of the reactive oxygen species that appear commonly on the human face is hydrogen peroxide. However, GPX7 can catalyze the reaction to reduce hydrogen peroxide to water and prevent the oxidation and aging of the skin [2],[3]. This is a way to nourish the skin in addition to the prevention of acne.

Caf1-AMP is a protein to captures and suppresses P. acne, our identified main pathogen for acne formation. In short, Caf1-AMP, after entering the pores on human faces at a relatively warm temperature, can cool down to repolymerize into a net-like structure which limits the movement of P. acne. The antimicrobial peptides attached to the protein can selectively kill the bacteria by disrupting the cell membrane.

TurboID-FGB based was developed based on the part design of last year’s BS_United_China 2022 [4]. TurboID-FGB is a protein ligase that binds biotin in around 10 minutes [5]. Through the binding of TurboID to a specific protein’s signal peptide that belongs to P. acne, biotinylation occurs to bind biotin onto a protein (specifically its lysine residues). Alongside streptavidin-phycoerythrin [6], we successfully inhibit the quorum-sensing mechanism of bacteria. When TurboID binds to the signal peptides of a protein, ATP and biotin are catalyzed together to produce biotinyl-5'-AMP. Using the energy from ATP, the product of biotnyl-5'-AMP can label selected proteins. The biotin bound to the protein then accumulates and effectively blocks the receptors of bacteria P. acne. Streptavidin-phycoerythrin can also join the biotin, creating an even larger barrier that blocks receptors and enabling a double-blocking system for quorum sensing. Consequently, it will be difficult for P. acne to communicate and undergo normal functions, such as virulence factor production and pathogenic behaviors [7].

Characterization of TurboID-FGB

Fig. 1. Staining Result of P. acne by Streptavidin-568 Binding to Activated Biotin through Oil Immersion.
We can notice that TurboID-FGB can successfully label activated biotin onto P. acne, as the red Streptavidin-568 can strongly bind to activated biotin and colors the P. acne which is labeled with abundant biotin.


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]


References

[1]GPX7 glutathione peroxidase 7 [Homo sapiens (human)] - Gene - NCBI. (n.d.). Retrieved from www.ncbi.nlm.nih.gov website: https://www.ncbi.nlm.nih.gov/gene/2882
[2]Rinnerthaler, M., Bischof, J., Streubel, M., Trost, A., & Richter, K. (2015). Oxidative Stress in Aging Human Skin. Biomolecules, 5(2), 545–589. https://doi.org/10.3390/biom5020545
[3]UniProt. (n.d.). Retrieved October 4, 2023, from www.uniprot.org website: https://www.uniprot.org/uniprotkb/Q96SL4/entry
[4]“Project Description.” | BS_United_China - iGEM 2022, 2022.igem.wiki/bs-united-china/description. Accessed 3 Oct. 2023.
[5]Cho, Kelvin F., et al. “Proximity labeling in mammalian cells with turboid and split-turboid.” Nature Protocols, vol. 15, no. 12, 2 Nov. 2020, pp. 3971–3999, https://doi.org/10.1038/s41596-020-0399-0.
[6]“PE Streptavidin.” BioLegend, www.biolegend.com/ja-jp/products/pe-streptavidin-1475?GroupID=GROUP23. Accessed 4 Oct. 2023.
[7]Rutherford, Steven T, and Bonnie L Bassler. “Bacterial quorum sensing: its role in virulence and possibilities for its control.” Cold Spring Harbor perspectives in medicine vol. 2,11 a012427. 1 Nov. 2012, doi:10.1101/cshperspect.a01242