Team:Harvard/Description
Project Description
Our team will focus on developing antimicrobial biofilms and bringing production to industrial relevancy. Our project offers the potential to make the biological manufacture of antimicrobial peptides an economically feasible alternative to chemical synthesis.
The first part of our project, functionalizing biofilms, will involve attaching functional proteins to curli fibers, the main proteinaceous component of biofilms naturally produced by Escherichia coli. In anticipation of the potential obstacles of synthesizing AMPs in a microbe, we will be using a library approach to cast a wide net on the range of documented AMPs. Through our screen of a library of AMP and curli fiber fusions, we hope to identify characteristics of AMP structure that make them more amenable to maintaining structure and function when fused to curli fibers, expressed, and secreted in E. coli. This can then act as a starting point for identifying or designing additional AMPs for functionalizing curli.
The second part of our project, bringing production to industrial relevancy, can be further broken down into three aspects: pathway optimization, resource management, and bioprocess optimization. The first of those, pathway optimization, will involve finding the optimal expression ratios of the various proteins involved in the curli production pathway. This is to minimize resource and energy wasted in the production of unused chaperone or membrane proteins. The second, resource management, will focus on diverting as much of the cell’s resources and energy to the curli production pathway while still maintaining cell viability. We will be using a strain of E. coli already optimized for recombinant protein production, BL21 (DE3), but we will also test our system with other knockouts to further optimize the strain for our purposes. Lastly, the bioprocess optimization aspect will involve the development of a bioreactor that not only provides a larger fermentation environment, but also integrates the extraction and filtration of the biofilm from the cells in a smooth and continuous process.
