Difference between revisions of "Part:BBa K5115002"
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===Introduction===
 
===Introduction===
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Hpn encodes a nickel-binding protein crucial for maintaining nickel homeostasis in ''Helicobacter pylori''. With its high histidine content, Hpn plays a vital role in sequestering nickel ions, preventing toxicity while ensuring proper nickel transfer to urease. This process is essential for the activation of urease, an enzyme that enables ''H. pylori'' to colonize the acidic gastric environment.<ref>Vinella, D., Fischer, F., Vorontsov, E., Gallaud, J., Malosse, C., Michel, V., Cavazza, C., Robbe-Saule, M., Richaud, P., Chamot-Rooke, J., Brochier-Armanet, C., & De Reuse, H. (2015). Evolution of Helicobacter: Acquisition by Gastric Species of Two Histidine-Rich Proteins Essential for Colonization. PLoS pathogens, 11(12), e1005312.</ref>.
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The EP sequence encodes an endogenous encapsulation peptide, which plays a crucial role in directing external proteins into bacterial microcompartments like carboxysomes. This targeting mechanism is essential for protein encapsulation within these structures, aiding in the assembly of a functional, proteinaceous shell that sequesters enzymes or other proteins, ensuring efficient catalysis or protection from environmental stress.
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<ref>Li, T., Jiang, Q., Huang, J., Aitchison, C. M., Huang, F., Yang, M., Dykes, G. F., He, H. L., Wang, Q., Sprick, R. S., Cooper, A. I., & Liu, L. N. (2020). Reprogramming bacterial protein organelles as a nanoreactor for hydrogen production. Nature communications, 11(1), 5448. </ref>.
  
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K5115000 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K5115002 SequenceAndFeatures</partinfo>
  
  

Revision as of 09:17, 15 September 2024


EP

contributed by Fudan iGEM 2023

Introduction

The EP sequence encodes an endogenous encapsulation peptide, which plays a crucial role in directing external proteins into bacterial microcompartments like carboxysomes. This targeting mechanism is essential for protein encapsulation within these structures, aiding in the assembly of a functional, proteinaceous shell that sequesters enzymes or other proteins, ensuring efficient catalysis or protection from environmental stress. [1].

contributed by Fudan iGEM 2023
Figure 1. The biosynthetic pathway of shinorine, porphyra-334, palythine-Ser, and palythine-Thr

Usage and Biology

We performed codon optimization specifically for the Escherichia coli K12 strain, resulting in the creation of this part. The Hpn protein binds absorbed nickel ions, enhancing E. coli's tolerance to elevated nickel concentrations within E. coli.

Characterization

Nickel tolerance Assay

We.

contributed by Fudan iGEM 2023
Figure 2. Anti-UV Assay.

Our experiments demonstrated that expressing Hpn in E. coli significantly enhanced the bacterium's ability to absorb nickel ions. We hypothesize that this increased absorption is due to Hpn’s strong nickel-binding capacity. However, for optimal nickel uptake, other factors, such as the availability of additional nickel transporters or regulatory proteins, may also be necessary.

contributed by Fudan iGEM 2023
Figure 3. Plates displaying transformed E. coli after anti-UV assay.


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 646
  • 1000
    COMPATIBLE WITH RFC[1000]


References

  1. ↑ Li, T., Jiang, Q., Huang, J., Aitchison, C. M., Huang, F., Yang, M., Dykes, G. F., He, H. L., Wang, Q., Sprick, R. S., Cooper, A. I., & Liu, L. N. (2020). Reprogramming bacterial protein organelles as a nanoreactor for hydrogen production. Nature communications, 11(1), 5448.