Coding

Part:BBa_K5208003

Designed by: Yuqi Fu   Group: iGEM24_Hangzhou-BioX   (2024-09-23)


AiiM, N-acyl homoserine lactonase from Microbacterium testaceum StLB037

AiiM encodes for an N-acyl homoserine lactonase in M. testaceum StLB037. This enzyme catalyzes the hydrolysis of N-Acyl homoserine lactone (AHL) (Wang et al., 2010).

Figure 1. Degradation of N-acyl homoserine lactone (AHL) by AiiM. The dashed line on the left shows where the enzyme breaks the bond (Chen et al., 2013; Dong et al., 2000).


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 516
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 81
    Illegal BamHI site found at 575
    Illegal XhoI site found at 526
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 482
    Illegal BsaI site found at 610
    Illegal SapI site found at 297



Usage and Biology

AiiM degrades AHLs, a class of signaling molecules that induce quorum sensing. Quorum sensing triggers the expression of virulence factors in pathogens, including Aeromonas hydrophila, an aquatic bacterium that threatens aquaculture (Coquant et al., 2020; Hlordzi et al., 2020). High expression of AiiM in the probiotic Bacillus subtilis reduces the virulence of A. hydrophila through quorum quenching and saves aquatic animals.

Figure 2. Gel electrophoresis of PCR product of AiiM.

References

Chen, F., Gao, Y., Chen, X., Yu, Z., & Li, X. (2013). Quorum quenching enzymes and their application in degrading signal molecules to block quorum sensing-dependent infection. Int J Mol Sci, 14(9), 17477-17500. https://doi.org/10.3390/ijms140917477

Coquant, G., Grill, J. P., & Seksik, P. (2020). Impact of N-Acyl-Homoserine Lactones, Quorum Sensing Molecules, on Gut Immunity. Front Immunol, 11, 1827. https://doi.org/10.3389/fimmu.2020.01827

Dong, Y. H., Xu, J. L., Li, X. Z., & Zhang, L. H. (2000). AiiA, an enzyme that inactivates the acylhomoserine lactone quorum-sensing signal and attenuates the virulence of Erwinia carotovora. Proc Natl Acad Sci U S A, 97(7), 3526-3531. https://doi.org/10.1073/pnas.97.7.3526

Hlordzi, V., Kuebutornye, F. K. A., Afriyie, G., Abarike, E. D., Lu, Y., Chi, S., & Anokyewaa, M. A. (2020). The use of Bacillus species in maintenance of water quality in aquaculture: A review. Aquaculture Reports, 18, 100503. https://doi.org/https://doi.org/10.1016/j.aqrep.2020.100503

Wang, W. Z., Morohoshi, T., Ikenoya, M., Someya, N., & Ikeda, T. (2010). AiiM, a novel class of N-acylhomoserine lactonase from the leaf-associated bacterium Microbacterium testaceum. Appl Environ Microbiol, 76(8), 2524-2530. https://doi.org/10.1128/AEM.02738-09

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