Part: PRhl_31_CheZ_J23117_34_RhlR

Bba_K2594011

The plasmid senses and responds to N-butanoyl-L-Homoserine Lactone (C4-HSL), a quorum sensing molecule that is present in Pseudomonas aeruginosa, a common component of biofilms. The J23117 promoter constitutively expresses RhlR, which activates the PRhl promoter site when bound to C4-HSL. Once quorum sensing signals are detected, the production of CheZ, a protein responsible for chemotactic ability, begins. As the plasmid senses C4-HSL, the microbe begins moving in a straight-line path along the concentration gradient. If the microbe leaves the concentration gradient, it will begin tumbling again until it enters back into the gradient, eventually arriving at the biofilm.

Part: Chitinase B4A

Bba_K2594013

This plasmid is engineered to express a chitinase originated from Serratia marcescens B4A fused to the N- and C-term domains of Pseudomonas syringae ice-nucleation protein. The fusion protein will be anchored to the Escherichia coli cell surface. Chitinase is an enzyme that degrades chitin, a major component in the structure of cell walls for plant and fungi cells. In our project, the first plasmid PRhl_31_CheZ_J23117_34_RhlR allows the E. Coli to swim next to the biofilm with ice-nucleating protein-Chitinase B4A fusion protein on the surface. The degradation of the cell wall allows cinnamaldehyde easier access to the cell membrane and ultimately the destruction and death of the cell. Since the ice-nucleation protein keeps Chitinase B4A anchored to the cell membrane of the engineered E. Coli, lower concentrations of the chitinase will be required to effectively damage and destroy the biofilm.

Part: Chitinase C-1

Bba_K2594012

This plasmid is engineered to express the gene chiC1 fused to the N- and C-term domains of Pseudomonas syringae ice-nucleation protein. The fusion protein will be anchored to theEscherichia coli cell surface. Chitinase is an enzyme that degrades chitin, a major component in the structure of cell walls for plant and fungi cells. In our project, the first plasmid PRhl_31_CheZ_J23117_34_RhlR allows the E. Coli to swim next to the biofilm with ice-nucleating protein-Chitinase C-1 fusion protein on the surface. The degradation of the cell wall allows cinnamaldehyde easier access to the cell membrane and ultimately the destruction and death of the cell. Since the ice-nucleation protein keeps Chitinase C-1 anchored to the cell membrane of the engineered E. Coli, lower concentrations of the chitinase will be required to effectively damage and destroy the biofilm.