Team:Stanford-Brown/SB16 BioMembrane Nylon

The Problem

Nylon-6 is industrially produced through the chain-growth polymerization of caprolactam, the cyclical form of 6-aminocaproic acid (6-ACA). Our team investigated potential biosynthesis routes for this monomer, looking for pathways with common starting substrates to design a production system for 6-ACA that could be implemented on an extraterrestrial body. Past retrosynthetic analyses have uncovered two fermentative pathways for the production of 6-ACA, which both lacked required biocatalytic steps until 2015. [1][2] Last October, researcher Stefan Turk published a study detailing and potential candidates enzymes for these steps and characterizing their activity in vivo in E. coli. [3] With the novel nature of his work in mind, our team contacted Turk in request of a sample of his engineered strain eAKP-672 with the highest 6-ACA yield when grown in glucose containing medium. We hoped to adjust Turk's synthetic plasmid designs and/or make our own genomic edits to optimize the metabolic flux of his biosynthetic pathways. Turk replied in the affirmative, shipping us two separate cell samples containing plasmids pAKP-96 and pAKP-444, which held the six enzymes necessary to induce one of the two above mentioned pathways in E. coli.