Composite
Dsb-Art175

Part:BBa_K1659002

Designed by: Wei Chung Kong   Group: iGEM15_Oxford   (2015-08-26)
Revision as of 12:46, 26 August 2015 by Registry (Talk | contribs)

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Artilysin Art-175 fused at N-terminal with DsbA signal peptide

Artilysins are an exciting class of enzyme-based antibacterials. Their name is derived from "artificial endolysin" and they exploit the lytic power of bacteriophage-encoded endolysins. Endolysins are peptidoglycan hydrolases produced at the end of the lytic cycle that pass through the cytoplasmic membrane, degrade the peptidoglycan layer and cause the osmotic lysis of the infected bacterial cell, thus liberating the progeny. Purified endolysins have been used to kill Gram-positive pathogens. Gram-negative bacteria, however, have a protective outer membrane containing lipopolysaccharide (LPS) that serves as a barrier against the peptidoglycan hydrolytic activity of endolysins from the outside.

To overcome this problem, selected polycationic or amphipathic peptides that locally destabilize the LPS layer have been covalently fused to endolysins. Biers et al. fused the sheep myeloid antimicrobial peptide (SMAP-29) to the N terminus of the endolysin KZ144 to create Artilysin Art-175. Art-175 has been shown to be a highly potent antibacterial that acts in minutes to kill virtually all Pseudomonas aeruginosa strains. [2]

BBa_K1659002 is a composite of Art-175 with the N-terminal DsbA signal peptide with a hexahistidine tag. DsbA targets recombinant proteins to the co-translational signal- recognition-particle (SRP)-dependent pathway to mediate secretion out of the bacterial chassis. [2]


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 417
    Illegal AgeI site found at 616
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//collections/antimicrobial
Parameters
None