Parts

Overview

Our components were designed in a modular manner, which not only facilitates usage by our own team but also by other teams with similar objectives. To mitigate air pollution caused by straw burning, we innovatively adopted a biosynthetic approach to utilize lignin for the production of acetaminophen (AAP)—a high-value compound that exhibits excellent application potential in cold treatment and analgesia. We constructed an artificial synthetic pathway by utilizing several endogenous genes of Escherichia coli and exogenously introducing multiple genes derived from Agaricus bisporus (button mushroom) and Pseudomonas aeruginosa. After evaluating the catalytic efficiency of several isozymes, the ABH60 gene and PANAT gene were ultimately selected. Additionally, the production of irrelevant by-products was reduced through the knockout of the endogenous nhoA gene and the rational application of the I38 promoter. Furthermore, we drew on the designs and experiences of previous iGEM teams, which simplified our experimental procedures and improved our overall results. Modular design is a core principle of synthetic biology, enabling production systems to generate entirely different outcomes with only minor modifications. This approach not only contributed to the significant success of our project but also allows for the rapid adaptation of our method to the production of other homologous compounds through module exchange.This framework will be particularly beneficial to future iGEM teams by providing design insights and key components. It aligns with the iGEM spirit of fairness, collaboration, and mutual learning. We anticipate that our PARTS will benefit numerous teams, further demonstrating the meaningful impact of our work.

New Composite Part

In our project, we constructed a novel composite biological part, pabABC. In nature, the PabABC enzyme complex is known to catalyze the conversion of chorismic acid to p-aminobenzoic acid (p-ABA). In our engineered pathway, we proposed to utilize this PabABC enzyme to instead utilize vanillic acid as a non-natural substrate for its conversion to p-ABA. Our experimental results successfully confirmed that the PabABC enzyme can indeed catalyze this novel transformation, representing a breakthrough step in our work. This pivotal finding validates the overall feasibility of our proposed pathway.

This achievement not only demonstrates the application potential of vanillic acid as a lignin-derived compound in biosynthesis but also provides a new strategy for the green production of high-value pharmaceutical molecules.