Glucuronic Acid Bioproduction Engineering Service

Glucuronic Acid (GlcA) is a carbohydrate acid vital in detoxification and drug metabolism in humans, used in the food and pharmaceutical industries. Industrial production is typically enzymatic or fermentation-based. However, the enzymatic route using Glucose Oxidase (GOx) and Uronate Dehydrogenase suffers from low conversion rate in enzyme synthesis due to enzyme instability and substrate/product inhibition. Fermentation often leads to many by-products in fermentation (e.g., gluconic acid, oxalic acid), complicating purification and reducing yield. A highly specific, high-yield biological route is needed.

CD Biosynsis offers a synthetic biology service focused on highly efficient and specific GlcA production. Our core strategy involves directed evolution of Aspergillus niger Glucose Oxidase (GOx) . We engineer GOx variants with enhanced thermal stability, higher catalytic efficiency, and improved selectivity toward the Glucuronic Acid pathway intermediates. This is coupled with the optimization of fermentation conditions , specifically fine-tuning pH, temperature, and dissolved oxygen levels to favor the action of the engineered GOx and minimize side reactions and byproduct formation. This integrated approach aims to deliver a high-yield, high-purity, and cost-effective Glucuronic Acid product.

Pain Points

Developing a robust Glucuronic Acid production process faces these key challenges:

• Low Conversion Rate in Enzyme Synthesis: Enzymes involved in GlcA synthesis often exhibit poor catalytic efficiency (k}_{\text{cat}}/\text{K}_{\text{m}) and are highly susceptible to substrate/product inhibition , limiting the total conversion achieved.
• Many By-products in Fermentation: Non-specific oxidation and metabolic leakage in microbial hosts lead to the co-production of gluconic acid, xylonic acid, and other organic acids , significantly reducing GlcA purity and complicating separation.
• Enzyme Stability: Glucose Oxidase (GOx) and other key enzymes can exhibit poor thermal or pH stability , necessitating strict and often costly reaction controls.
• Oxygen Dependence: GOx-based reactions require high levels of dissolved oxygen (DO) , which is difficult to maintain at industrial scale without incurring high aeration costs.

An effective solution must boost enzyme performance, maximize GlcA specificity, and simplify downstream processing.

Solutions

CD Biosynsis utilizes advanced enzyme and fermentation engineering to optimize Glucuronic Acid production:

Directed Evolution of Aspergillus niger Glucose Oxidase

           

We perform high-throughput screening of GOx libraries to identify variants with significantly higher activity and improved stability under industrial conditions.

Optimization of Fermentation Conditions

We fine-tune pH, temperature, and aeration strategies to create an environment that maximizes the conversion efficiency of the target pathway and suppresses side reactions.

Byproduct Pathway Knockout

In fermentation hosts (if used), we utilize CRISPR technology to delete or downregulate metabolic pathways leading to non-target organic acids (e.g., oxalate), ensuring high purity.

Co-factor Recycling Enhancement

We integrate and optimize systems for efficient NAD}^{+ regeneration within the host or enzymatic system, ensuring the continuous operation of Uronate Dehydrogenase.

This systematic approach focuses on improving the catalytic core (GOx) and optimizing the surrounding reaction environment for high specificity and conversion.

Advantages

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

Enhanced Catalytic Efficiency

Directed evolution yields GOx variants with higher reaction speeds (k}_{\text{cat}) and improved affinity for the substrate, boosting conversion rate.

High Product Purity

Targeted knockouts and optimal conditions significantly minimize byproduct formation , simplifying costly downstream purification steps.

Operational Stability

Engineered enzymes exhibit better tolerance to temperature, pH variation , and substrate concentration, allowing for more robust industrial processes. [Image of Cost Reduction Icon]

Reduced Aeration Costs

If GOx is integrated into a microbial host, balanced O}_2$ consumption/supply can be achieved, potentially lowering the energy required for large-scale aeration .

Sustainable Production Route

Glucuronic Acid is produced from simple, renewable glucose , offering a green alternative to chemical routes.

We provide a specialized platform for the high-specificity and high-yield production of Glucuronic Acid.

Process

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

• GOx Library Generation: Create diverse libraries of the A. niger GOx gene using random mutagenesis or saturation mutagenesis techniques.
• High-Throughput Screening: Implement a sensitive, high-throughput assay to screen thousands of variants for increased catalytic activity and stability toward the GlcA pathway.
• Pathway Integration (if microbial): If using a microbial host, integrate the improved GOx variant and the necessary uronate dehydrogenase into the genome for stable expression.
• Fermentation Condition Optimization: Systematically adjust key parameters (e.g., DO level, pH control, nutrient feeding ) in small-scale bioreactors to maximize GlcA yield and purity.
• Scale-Up Validation: Validate the final engineered system (enzyme or host) at the pilot scale to confirm industrial viability and robustness.
• Result Report Output: Compile a detailed Experimental Report including enzyme kinetics (k}_{\text{cat}, K}_{\text{m}), stability data, and final process metrics (titer, yield, and purity) .

Technical communication is maintained throughout the process, focusing on timely feedback regarding enzyme performance and product specificity.

Explore the potential for a high-performance, high-purity Glucuronic Acid supply. CD Biosynsis provides customized production solutions:

• Detailed Enzyme Kinetic and Stability Report , showing the performance advantages of the directed evolution variants.
• Consultation on downstream purification protocols tailored to the high-purity product stream.
• Experimental reports include complete raw data on glucose conversion efficiency and byproduct profile , essential for commercial application.