Coding

Part:BBa_K2018015:Design

Designed by: Brian Kenn Baltzar   Group: iGEM16_SDU-Denmark   (2016-06-05)


Lacticin Q-Lacticin Z producing


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This hybrid bacteriocin were designed by taking the individual bacteriocin genes, putting them together with a Gly-Gly-Gly hinge in between. The genes were found in the NCBI gene database. The reason for the Glycin hinge in between the two bacteriocins is that it should function as a flexible domain, giving each bacteriocin the possibility to maintain their function.

Why? Our thought behind this hybrid were that maybe two bacteriocins would have a synergistic effect and work together better and more specific than they would on their own.

Literature shows that MIC of Lacticin Q (as a single bacteriocin protein) is 1,800 nM against Staphylococcus aureus:ATCC 12600((Fujita et al. 2007) , in our MIC tests we showed that the hybrid bacteriocin inhibited growth of S. aureus:CC398, USA300 & hVISA at a concentration as low as 642 nM. It even showed antimicrobial effect against Pseudomonas aeruginosa:PAO1 with a MIC value of 1283 nM, an effect that (to our knowledge) neither of the two single bacteriocins has showed effect against.

Source

NCBI genebank database

lacticin Z: Accession: AB298287.1

lacticin Q:Accession: AB235201.1

References

  • Fujita, Koji, Shiro Ichimasa, Takeshi Zendo, Shoko Koga, Fuminori Yoneyama, Jiro Na- kayama og Kenji Sonomoto (2007). Structural Analysis and Characterization of Lacticin Q, a Novel Bacteriocin Belonging to a New Family of Unmodifed Bacteriocins of Gram-Positive Bacteria". I: Applied and Environmental Microbiology 73.9, s. 287