Coding

Part:BBa_K5208004

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


AhlD, N-acyl homoserine lactonase from Arthrobacter sp. IBN110

AhlD encodes for an N-acyl homoserine lactonase in Arthrobacter sp. IBN110. This enzyme catalyzes the hydrolysis of N-Acyl homoserine lactone (AHL) (Park et al., 2003).

Figure 1. Degradation of N-acyl homoserine lactone (AHL) by AhlD. 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
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 614
    Illegal BamHI site found at 728
    Illegal XhoI site found at 313
    Illegal XhoI site found at 427
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 310
    Illegal BsaI.rc site found at 34



Usage and Biology

AhlD 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 AhlD in the probiotic Bacillus subtilis reduces the virulence of A. hydrophila through quorum quenching and saves aquatic animals.

Figure 2. Gel electrophoresis of colony PCR products for verification of correct transformation of plasmid pHT43-AhlD into B. subtilis WB600.

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

Park, S. Y., Lee, S. J., Oh, T. K., Oh, J. W., Koo, B. T., Yum, D. Y., & Lee, J. K. (2003). AhlD, an N-acylhomoserine lactonase in Arthrobacter sp., and predicted homologues in other bacteria. Microbiology (Reading), 149(Pt 6), 1541-1550. https://doi.org/10.1099/mic.0.26269-0

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