Contribution

Overview

As a leading global platform for synthetic biology competitions, iGEM has significantly contributed to the rapid development of this field. The platform fosters international knowledge exchange and technological collaboration through promoting teams to learn from and build upon the innovative experiences of the world's top research groups. To better integrate into this community, we have shared our valuable experiences from the competition on the iGEM platform, including contributions in areas such as experimental design and project implementation. We hope these insights will inspire and assist future researchers in advancing the future of synthetic biology.

I. Contributions to the iGEM Community

1. Provide reusable standardized biological components:

This project has successfully constructed a series of plasmids, including pYB1a-pabABC (the pathway from vanillic acid to p-aminobenzoic acid), pYB1a-pabABC-ABH60 (efficiently converting vanillic acid to p-aminophenol), and pSB1c-I38-PANAT (optimizing the conversion from p-aminophenol to acetaminophen). All components comply with the iGEM standard biological part specifications and can be uploaded to the iGEM Parts Registry. Subsequent teams can directly use these components to quickly build aromatic drug synthesis pathways, reducing the time cost of repeated construction.

2. Share the complete experimental protocol and data:

The full-chain fermentation parameters verified in the project for "vanillic acid → p-aminobenzoic acid (p-ABA) → p-aminophenol (p-AP) → acetaminophen (AAP)" (such as 10% glycerol as the optimal carbon source, 15 mM L-glutamine to improve substrate conversion rate, 250 mL conical flask to optimize dissolved oxygen level), HPLC detection methods (p-ABA peak time at 5 min, vanillic acid at 12 min) will be made public in the form of a wiki. This will provide a reproducible experimental template for similar research.

3. Propose an innovative research paradigm of "industrial waste → drugs":

For the first time in iGEM, the synthesis of AAP using a lignin degradation product (vanillic acid) as a raw material was achieved, breaking the dependence on traditional petrochemical routes. This provides new research ideas for the direction of "environmentally friendly drug synthesis" within the community and promotes more teams to pay attention to the interdisciplinary field of "waste valorization + pharmaceutical production".

II. Contributions to the field of synthetic biology

1. Filling the gap in metabolic pathways from lignin to aromatic drugs:

By overexpressing the gene pabABC (catalyzing the conversion of vanillic acid to p-ABA) in Escherichia coli, heterologously expressing ABH60 (replacing MNX1 to increase p-AP production with an efficiency more than 3 times that of MNX1), and optimizing PANAT to replace nhoA to reduce side reactions, a complete synthetic pathway from lignin derivatives to acetaminophen precursors was constructed for the first time. This provides a new metabolic engineering strategy for synthetic biology in "synthesizing high-value chemicals using renewable resources".

2. Develop efficient methods for strain modification and regulation:

The synergistic strategy of "nhoA gene knockout + dual-plasmid co-expression" was verified (the yield of the knockout strain p-AP was significantly higher than that of the interference strain), as well as the precise regulation effect of the temperature-sensitive promoter I38. This provides a reference modification scheme for "pathway enhancement + bypass blocking" in microbial cell factories, and helps synthetic biology achieve technological breakthroughs in the field of metabolic flux optimization.

III. Contributions to Society

1. Promote the green transformation of the pharmaceutical supply chain:

The traditional production of acetaminophen relies on fossil raw materials such as phenol and benzene, requiring high-temperature and high-pressure conditions and generating toxic waste. This research project uses industrial waste lignin (with a global annual emission of 50-70 million tons and a utilization rate of less than 2%) as the raw material to synthesize pharmaceutical precursors under mild conditions of 30 ℃. This can reduce the consumption of petrochemical resources and environmental pollution, alleviate the dependence of the global pharmaceutical supply chain on fossil fuels, and contribute to the achievement of the "dual carbon" goals.

2. Enhancing the capacity of public health security:

Acetaminophen, recognized by the WHO as an essential medicine, has a global annual output of 200,000 tons, but its production is highly concentrated, making it prone to supply shortages. The technology of this project can integrate drug production with lignin-producing areas (such as pulp mills and biorefineries) to achieve "local sourcing and local production". This reduces drug transportation costs and supply risks, and in particular, provides the possibility of independent pharmaceutical production for regions lacking petrochemical resources but rich in lignin, thereby enhancing public health emergency response capabilities.

3. Creating synergistic value between the economy and the environment:

Currently, the value of lignin when burned as fuel is only $0.18 per kilogram. After converting it into drug precursors through this research project, its value can be increased to $1.08 per kilogram. This not only creates new sources of income for industries such as pulp and biorefining but also reduces carbon emissions from the incineration of industrial waste, achieving a win-win situation of "economic benefits + environmental governance" and promoting the development of a circular bioeconomy.