Overview

Our project aims to leverage synthetic biology to promote the genetic engineering of a single E. coli culture system. This system is designed to utilize the enzyme KerDZ to break down keratin from textile waste into valuable amino acids. Additionally, it would incorporate the enzyme CelCD to further process the cellulose found in textiles, into a carbon source for microbial feedstock. The amino acids and feedstock would then be re-used by our E. coli to biosynthesize spider silk, a material with significant potential for sustainable applications. In our dry lab work, we concentrated on designing and optimizing the genetic constructs and expression systems to facilitate the degradation of keratin and cellulose and the production of spider silk.

Parts Designations

Biobrick Parts

Our Favourite Part: RBS 1_12000

The Ribosome Binding Site (RBS) is an essential sequence that plays a critical role in translation initiation in prokaryotes, facilitating efficient protein synthesis. RBS 1_12000, along with variants RBS 2_220, 3_126, and 4_59, were originally characterized for its ability to work alongside a T7 inducible promoter to regulate the expression of a synthetic operon containing the bphS, bphO, and yhjH genes. Each RBS variant was tested for its efficiency in controlling and regulating this synthetic operon.

Through extensive testing, RBS 1_12000 demonstrated the best performance overall, showing improved growth conditions and significantly higher fluorescence production compared to the other variants. This made it the optimal choice for our key constructs, such as pTac_RBS1_SpiderSilk_sfGLP_pJUMP24 and J23119_RBS1_KerDZ_pJUMP28. These constructs are fundamental to our project, and the use of RBS 1_12000 has contributed to their effectiveness in meeting our experimental goals. Below are images of the constructs made using RBS 1_12000.