Part:BBa_K3183017

CD27L Endolysin Targeting C. difficile Codon-Optimized for L. reuteri

This part contains CD27L, an endolysin specific to C. difficile cell walls, targeting it for lysis. This endolysin works by cleaving glycopeptides¹ (Fig. 1), breaking the cell's peptidoglycan and compromising the structural integrity of its cell wall. Two X-ray crystal structures were solved for the N-terminal² and C-terminal domains³. These are joined by a linker, which we've added ourselves (Figure 2).

Figure 2: X-ray Crystal Structure of CD27L; PDB codes 3QAY (green) & 4CU5 (purple) with a linker (yellow) drawn in with PyMol by Team Oxford 2019. Note the Zn²⁺ ion (silver) present in the putative active-site cleft, which may be involved with catalysis.

Figure 1: CD27L Catalytic Reaction.

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 719
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Use by Team Oxford 2019

This part was used in the composite part BBa_K3183103. Please see its documentation for more information.

References

1. Mayer, M. J., et al. “Molecular Characterization of a Clostridium Difficile Bacteriophage and Its Cloned Biologically Active Endolysin.” Journal of Bacteriology, vol. 190, no. 20, 2008, pp. 6734–6740., doi:10.1128/jb.00686-08.
2. Mayer, M. J et al. “Structure-based modification of a Clostridium difficile-targeting endolysin affects activity and host range.” Journal of bacteriology vol. 193,19 (2011): 5477-86. doi:10.1128/JB.00439-11
3. Dunne, Matthew et al. “The CD27L and CTP1L endolysins targeting Clostridia contain a built-in trigger and release factor.” PLoS pathogens vol. 10,7 e1004228. 24 Jul. 2014, doi:10.1371/journal.ppat.1004228