Adipic Acid Bioproduction Engineering Service

Adipic Acid (ADA) is a crucial dicarboxylic acid, primarily used as a monomer for synthesizing Nylon 6,6 and as a component in polyurethanes. Conventional industrial synthesis relies on petrochemical feedstocks (cyclohexane/benzene), a process notorious for high emissions in petrochemical synthesis , particularly the release of nitrous oxide (N2O), a potent greenhouse gas. Biosynthesis offers a green alternative but is hampered by incomplete biosynthetic pathways in native hosts and tight regulation, resulting in low conversion rates and industrial viability.

CD Biosynsis offers a synthetic biology service focused on achieving high-titer, sustainable ADA production. Our core strategy involves modification of Escherichia coli fatty acid metabolism pathway , rerouting the highly efficient fatty acid degradation (beta-oxidation) cycle to function in reverse for the synthesis of the C6 intermediate (Hexanoyl-CoA). This is coupled with the overexpression of key enzymes for adipic acid synthesis , specifically the reductase/oxidase cascade necessary to convert the C6 fatty acid intermediate into the final Adipic Acid product. This integrated approach aims to deliver a high-yield, pure, and environmentally sustainable Adipic Acid product, avoiding the N2O emissions of petrochemical routes.

Pain Points

Transitioning to competitive biosynthetic Adipic Acid production faces these key challenges:

• High Emissions in Petrochemical Synthesis: The industrial route generates large volumes of N2O , a greenhouse gas with a global warming potential 300 times that of CO_2, posing a major environmental burden.
• Incomplete Biosynthetic Pathways: Native hosts lack the full set of enzymes needed for de novo synthesis of ADA from sugar. Key steps, particularly the terminal oxidization of the C6 intermediate , are often missing or inefficient.
• Metabolic Leakage and Byproducts: The C6 intermediates, such as Hexanoyl-CoA, are prone to being consumed by native fatty acid pathways for biomass synthesis or converted into non-target molecules.
• Hydrophobicity Issues: The product or its intermediate precursors are highly hydrophobic, which can lead to intracellular toxicity and poor product secretion , limiting the final yield.

A successful solution must establish a complete, high-flux pathway for ADA synthesis and ensure its efficient export.

Solutions

CD Biosynsis utilizes advanced metabolic engineering to optimize Adipic Acid production in E. coli :

Modification of E. coli Fatty Acid Metabolism Pathway

           

We engineer the beta-oxidation pathway to operate in reverse (the reductive mode) for chain elongation, specifically focusing on the production of the C6 intermediate, Hexanoyl-CoA.

Overexpression of Key Enzymes for Adipic Acid Synthesis

We introduce and overexpress the heterologous enzymes (e.g., dehydrogenases and oxidoreductases ) required to convert Hexanoyl-CoA into the final product, Adipic Acid.

Competing Pathway Blockage

We use CRISPR technology to knock out native enzymes (thioesterases and degradation enzymes) that consume the Hexanoyl-CoA intermediate for other cellular functions.

Co-factor and Redox Balance Optimization

We tune the central metabolism (e.g., PPP) to ensure sufficient NADPH supply , which is essential for the high-yield operation of the fatty acid synthesis pathway in the reductive mode.

This systematic approach is focused on establishing a complete and highly specific pathway for ADA synthesis in the host.

Advantages

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

Zero Nitrous Oxide Emissions

Microbial synthesis eliminates the generation of N2O , making it a dramatically greener process than the conventional route.

Renewable Feedstock Utilization

The host uses low-cost sugars or biomass derivatives instead of non-renewable benzene/cyclohexane as the carbon source.

High Adipic Acid Titer and Purity

Targeted pathway construction minimizes side products, leading to a cleaner broth and simpler purification. [Image of Cost Reduction Icon]

Reduced Energy Consumption

Fermentation occurs under mild temperature and pressure, significantly reducing the overall energy footprint compared to petrochemical processes.

Flexible Biosynthetic Route

The fatty acid pathway can be tuned to produce other diacids or monomers (C}4$, C}5$, C}8$) by adjusting pathway enzymes.

We provide a sustainable and cost-effective biosynthetic platform for industrial Adipic Acid production.

Process

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

• Reverse beta-oxidation Pathway Establishment: Inactivate native beta-oxidation degradation genes (e.g., fadA, fadB) and heterologously express the enzymes needed to run the cycle in the reductive (synthesis) direction.
• Terminal C6 Conversion: Introduce and optimize the expression of a dehydrogenase-aldehyde dehydrogenase-oxidase cascade to convert Hexanoyl-CoA to Adipic Acid .
• Precursor Supply Optimization: Engineer the central carbon metabolism to boost the supply of Acetyl-CoA and NADPH , the essential inputs for the reverse beta-oxidation pathway.
• Product Export Engineering: Screen or introduce specific dicarboxylic acid transporters to actively export the Adipic Acid from the cell, mitigating toxicity.
• Fermentation Performance Validation: Test the final engineered strain in fed-batch fermentation to assess Adipic Acid titer, yield, and process stability .
• Result Report Output: Compile a detailed Experimental Report including gene modification data, flux analysis, and fermentation metrics (yield, titer, and purity) , supporting process transfer.

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

Explore the potential for a high-performance, green Adipic Acid supply. CD Biosynsis provides customized strain engineering solutions:

• Detailed Metabolic Flux and Titer Analysis Report , demonstrating success in carbon redirection and titer increase.
• Consultation on fermentation control strategies optimized for NADPH regeneration and high-density growth.
• Experimental reports include complete raw data on carbon yield (g Adipic Acid}/\text{g sugar) and product purity , essential for commercial application.