Proof Of Concept

This biofilm is mainly made up of mono and polysaccharides. In Cutibacterium Acne, major components that make up the biofilm include proteins, lipids, and sugars, which all help to maintain the structural integrity of the biofilm. Since the biofilm 50% made out of polysaccharides, our goal is to target the the degradation pathway of polysaccharides in our gram-negative bacteria in hopes to reducing the biofilm. By targeting an enzyme known as PFK1 which is vital in all forms of life and catalyzes an irreversible step, we believe we are able to effectively damage the bacteria’s ability to produce biofilm, allowing antibiotics and antimicrobials to work more effectively or be administered. Biofilm is made when the bacterial cells adhere to a surface and produce a matrix of Extracellular Polymeric Substance (EPS). They become embedded in this matrix and it protects them from being destroyed. Therefore, before destroying any bacteria, the extracellular matrix around is necessary to be destroyed first. By focusing on PFK1, our team is able to break down the original biofilm formation and target the path of interest; this will be more effective compared to catalyzing a small reaction on the biofilm’s surface. To provide an example. proteinase K has been proven in studies to breakdown biofilms in other bacteria.

Our design was constructed using GenSmart. Using this technology we designed the insert and used the Pet28a+ backbone with Kanamycin resistance. Using our designed primers aka a portion of our basic part. We performed a PCR reaction and sequencing identifying our engineering success. The plasmid has an insert for PFK1, our enzymatic inhibitor and using this inhibitor we are able to test the effects of PFK1 on C. acnes based biofilms.

To come up with preventative measures against biofilm production in C.acnes.