Difference between revisions of "Part:BBa K5311006"

Revision as of 17:35, 29 September 2024

CAP 10-3 Endolysin

We hypothesized that the incorporation of an endolysin into our Cutibacterium acnes design would help to address both safety concerns and improve the release of the Cry proteins, maximizing therapeutic delivery. Since the bacteria will be in direct contact with the skin, using an endolysin enables controlled cell lysis, reducing the risks of having a GMO in prolonged contact with the skin. The RNA thermometer controls endolysin expression, activating it at the regular skin temperature (around 37ºC) to ensure precise timing, and facilitating the distribution of our product, as it wouldn’t start producing the endolysin until it reached the skin. Additionally, the endolysin helps facilitate the release of therapeutic Cry proteins, breaking down the bacterial cell walls and enhancing the effectiveness of the treatment against skin infestations. This dual function ensures safety while improving protein release.

Sequencing

Sequence and Features

Biology

In the case of the endolysin derived from the CAP 10-3 bacteriophage, its action specifically targets the degradation of the cell wall of Cutibacterium acnes, a Gram-positive bacterium with a cell wall rich in peptidoglycan, which is a crucial structural component. This endolysin is classified as an N-acetylmuramoyl-L-alanine amidase, which cleaves the bond between the N-acetylmuramic acid and the L-alanine residue in the peptidoglycan backbone.

By breaking down the peptidoglycan, the endolysin disrupts the rigid structure of the cell wall, leading to a loss of osmotic integrity. As a result, the bacterial cell can no longer withstand the internal pressure, causing rapid cell lysis. This mechanism is particularly effective against Gram-positive bacteria like C. acnes because they have a thick peptidoglycan layer that is more accessible to endolysins, unlike Gram-negative bacteria, which possess an additional outer membrane that restricts endolysin access.[1]

Advantages of Endolysins Against C. acnes:

• High Specificity: The CAP 3-10 Endolysin is known for its high specificity towards C. acnes, often recognizing unique peptidoglycan motifs present in the bacterial cell wall. This ensures that they selectively lyse the pathogenic bacteria without affecting other beneficial members of the skin microbiota [2].
• Low Risk of Resistance: Unlike traditional antibiotics, which typically target essential processes such as protein synthesis or DNA replication, endolysins act on the structural components of the bacterial cell wall, such as the peptidoglycan layer. The peptidoglycan is a highly conserved and essential component that is less prone to mutation, reducing the likelihood of bacteria developing resistance [3].
• Safety and reduced side effects: Endolysins have a favorable safety profile compared to traditional antibiotics, as they are proteins that are rapidly degraded in the human body without inducing toxic side effects. Furthermore, since they specifically target bacterial cell wall components, they do not interact with human cells, making them safe for topical or systemic applications [4].