Adipic Acid Precursor cis,cis-Muconic Acid Engineering Service

cis,cis-Muconic Acid is a critical bio-monomer precursor for Nylon 6,6 Monomer Adipic Acid, a foundational polymer for textiles and engineering plastics. The traditional method faces severe environmental and economic hurdles: Chemical synthesis from petrochemical benzene is highly energy-intensive and polluting. While biological routes exist, the biological yield is low, limiting economic viability.

CD Biosynsis offers a two-stage biocatalytic and metabolic engineering strategy to produce Muconic Acid and its final derivative Adipic Acid: Metabolic Engineering: Introduce a synthetic pathway to produce cis,cis-Muconic Acid from glucose via the Shikimate pathway in E. coli. This replaces the toxic petrochemical feedstock. We ensure the complete conversion to the final industrial monomer via Enzymatic Hydrogenation: Use Enoyl-CoA Reductase in a two-step chemo-enzymatic process for final conversion to Adipic Acid. This integrated approach provides a high-yield, sustainable route from sugar to Nylon precursor.

Pain Points

The sustainable production of Muconic Acid faces these main difficulties:

• Toxic Petrochemical Route: Conventional synthesis of Adipic Acid relies on benzene, a toxic and carcinogenic starting material, and nitric acid oxidation, resulting in the emission of the potent greenhouse gas N2O.
• Pathway Bottlenecks: The microbial synthesis of Muconic Acid from glucose is limited by the flux through the Shikimate pathway and the low activity of the key terminal enzyme catechol 1,2-dioxygenase or similar enzymes.
• Low Product Yield: Despite extensive research, the industrial yield of Muconic Acid in bio-based systems remains too low to compete economically with the established petrochemical process.
• Final Conversion: The Muconic Acid product must be hydrogenated to Adipic Acid, which traditionally requires harsh chemical hydrogenation steps and metallic catalysts.

A cost-effective solution must maximize the carbon flux from glucose and provide a green route for the final hydrogenation.

Solutions

CD Biosynsis utilizes integrated metabolic and enzymatic engineering to optimize Muconic Acid production:

Shikimate Pathway Flux Tuning

           

We engineer E. coli or yeast to enhance the precursor supply e.g. DAHP synthase overexpression through the Shikimate pathway, redirecting flux to the Muconic Acid synthetic branch.

Enzymatic Adipic Acid Conversion

We use a chemo-enzymatic cascade utilizing enzymes like Enoyl-CoA Reductase and a suitable co-factor system for the highly selective, mild-condition hydrogenation of Muconic Acid to Adipic Acid.

Terminal Enzyme Optimization

We optimize the expression and activity of the terminal enzymes that convert intermediates like catechol to cis,cis-Muconic Acid, ensuring maximum final titer and yield.

Bypass Feedback Inhibition

We implement enzyme detoxification and metabolic regulation to bypass the natural feedback inhibition present in the Shikimate pathway, ensuring continuous high flux.

This strategy provides a complete, sustainable, and highly efficient route from glucose to Nylon 6,6 precursor.

Advantages

Our Muconic Acid engineering service is dedicated to pursuing the following production goals:

Eliminate Toxic Petrochemicals Icon

Production from glucose avoids the use of toxic benzene and minimizes polluting N2O emissions.

High Efficiency Glucose Utilization Icon

Optimized Shikimate pathway allows for maximum carbon conversion from biomass-derived glucose.

Green Final Conversion Icon

The enzymatic hydrogenation step is highly selective and operates under mild conditions, replacing harsh chemical catalysts.

High Final Titer Achievement Icon

Flux tuning and feedback inhibition bypass lead to a significantly increased titer of Muconic Acid.

Bioreactor Scalability Icon

The engineered E. coli platform is a robust and well-established system for industrial fermentation scale-up.

We deliver a sustainable, high-performance solution for the bio-based production of Nylon 6,6 precursors.

Process

Our Muconic Acid strain engineering service follows a rigorous, multi-stage research workflow:

• Shikimate Pathway Engineering: Overexpress the key initial enzymes e.g. AroG}^{\text{fbr}}$ to relieve feedback inhibition and dramatically increase carbon flux through the Shikimate pathway.
• Synthetic Pathway Integration: Introduce the synthetic Muconic Acid pathway genes e.g. DAHPS, aroB}$, aroD}$, aroE}$, AroY}$, CatA}$ into the host E. coli}$.
• Terminal Enzyme Optimization: Screen and optimize the terminal pathway enzymes e.g. catechol dioxygenase for high specific activity and product yield to prevent metabolite accumulation.
• Fermentation and Muconic}$ Acid}$ Production: Develop high-cell-density fed-batch fermentation protocols to achieve maximum cis,cis-Muconic}$ Acid}$ titer and productivity.
• Adipic}$ Acid}$ Conversion Development: Establish the two-step chemo-enzymatic process combining high-yield Muconic Acid production with efficient, mild-condition Enoyl-CoA Reductase-based hydrogenation to final Adipic Acid.

Technical communication is maintained throughout the process, focusing on timely feedback regarding yield and product quality attributes.

Explore the potential for a sustainable, high-yield Adipic Acid supply. CD Biosynsis provides customized strain and process engineering solutions:

• Detailed Muconic Acid Titer and Yield Reports g/L, percentage of theoretical yield from optimized fermentation runs.
• Consultation on the integrated chemo-enzymatic conversion process to final Adipic Acid.
• Experimental reports include complete raw data on Shikimate pathway intermediate levels, byproduct formation, and final Adipic Acid purity.