Our team designed but was unable to test this module due to the limitation of not being able to work directly with Streptococcus Pneumoniae (Biosafety level 2) in our lab. This module proposes to kill Streptococcus Pneumoniae upon detection when coupled with the detection and regulation modules by releasing specific bacteriophage lytic enzymes, Cpl-1 and Pal.

Amidase (PAL)- cleaves the peptidoglycan between N-acetylmuramic acid and L-alanine

Lysozyme (CPl-1)- cleaves the glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine

Fig ? . The lysis Module.

The enzymes are tagged with osmY (Washington 2012) via a linker to be exported out of Escherichia coli. Both enzymes have very different N-terminal catalytic sites and share a similar C-terminal cell wall attachment site, which binds to choline in both cases. Cleavage with either of these enzymes results in a weakening in the cell wall, which leads to the externalization of the cytoplasmic membrane and ultimate lysis of S. pneumoniae.