Hyaluronic Acid (HA) Bioproduction Engineering Service

Hyaluronic Acid (HA) is a high-demand polymer crucial in medical (viscosupplementation, drug delivery) and cosmetic applications (moisturization). Traditionally produced by pathogenic Streptococcus strains (e.g., S. equi), HA bioproduction faces three critical industrial challenges. The first is operational: High viscosity of fermentation broth (especially above 4 g/L) severely limits oxygen mass transfer and mixing efficiency, constraining overall yield and efficiency. The second is safety: The traditional host carries the Risk of toxins and necessitates complex purification to eliminate microbial contaminants. The third is quality: HA's value is dependent on its high molecular weight (HMW). The Degradation of product molecular weight by native Hyaluronidase enzymes leads to reduced functional efficacy.

CD Biosynsis offers a comprehensive synthetic biology and metabolic engineering solution to address these bottlenecks. To stabilize the product quality, we implement Delete the hyaluronidase gene in the producing strain to prevent the HA molecular weight (MW) degradation. To eliminate the safety concern, we execute a Switch host to non-pathogenic industrial strains (e.g., Bacillus subtilis or engineered E. coli), making the final product safer and simplifying regulatory compliance and purification. To overcome the yield constraint caused by high viscosity and low polymerization rate, we apply Metabolic engineering . This involves a dual strategy: Balancing precursor (UDP-GlcNAc/UDP-GlcA) supply by upregulating key upstream pathway enzymes (e.g., GNA1, UGE) to ensure sufficient monomer availability, and overexpressing the hasA gene (Hyaluronate synthase) to increase the polymerization rate, allowing the production of high MW HA at high titers in the safer host.

Pain Points

Industrial HA production faces these key challenges:

• High Viscosity Constraint: HA is highly viscous. At HA concentrations above 4 g/L, the fermentation broth becomes highly non-Newtonian, leading to severe limitation of oxygen mass transfer and heat transfer, capping the maximum achievable yield.
• Risk of Toxins and Complex Purification: Using the traditional pathogenic host (Streptococcus sp.) inherently carries the risk of bacterial toxins (e.g., streptolysin O, exotoxins), requiring extremely complex, multi-stage, and costly purification steps to ensure product safety.
• Product Molecular Weight Degradation: Native Hyaluronidase enzymes (Hyaluronate lyases) present in the Streptococcus host actively degrade the HA polymer chain length , leading to a lower final MW and thus reducing the product's functional value.
• Precursor Supply Imbalance: Even with high Hyaluronate synthase activity, the conversion rate is limited by the imbalanced supply of the two necessary precursors, UDP-GlcNAc and UDP-GlcA.

A successful solution must remove toxins, stabilize MW, and overcome viscosity-related yield limits.

Solutions

CD Biosynsis utilizes advanced host and metabolic engineering to optimize HA production:

Switch Host to Non-Pathogenic Industrial Strains

           

We move the HA synthesis pathway (hasA and precursor genes) to safe, robust hosts like B. subtilis or E. coli, eliminating toxin risk.

Delete the Hyaluronidase Gene

We use CRISPR/Cas9 to knockout the endogenous hyaluronidase gene (e.g., hylA), ensuring HA MW stability and high functional quality.

Metabolic Engineering: Balance Precursor Supply

We use fine-tuned promoters to balance flux to UDP-GlcNAc and UDP-GlcA , maximizing HA synthase efficiency.

Overexpress the hasA Gene

We introduce an optimal hyaluronate synthase gene (hasA) on a strong inducible promoter to maximize the polymerization rate, overcoming yield constraints.

This systematic approach targets safety, MW stability, and production yield simultaneously.

Advantages

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

High Molecular Weight Stability

The Hyaluronidase gene deletion ensures the HA maintains its functional high MW throughout fermentation, addressing the degradation of product MW .

Enhanced Safety and Purity

Using non-pathogenic hosts eliminates the risk of toxins , simplifying purification and lowering Cost of Goods (COG).

High Production Titer Icon

Optimizing precursor flux and hasA overexpression maximizes the amount of HA produced per volume, compensating for viscosity constraints. [Image of Cost Reduction Icon]

Superior Biocompatibility Icon

Non-animal-derived HA produced in engineered bacteria is free from prion/viral risks, ideal for medical applications.

Robust Host Performance Icon

Industrial strains (E. coli, B. subtilis) are highly amenable to large-scale, high-density fermentation processes.

We provide a safer, more efficient, and quality-controlled platform for HA manufacturing.

Process

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

• Host Selection and Modification: Select a non-pathogenic host (E. coli or B. subtilis) and use CRISPR/Cas to delete the hyaluronidase gene if present.
• Pathway Construction: Introduce the HA synthase gene (hasA) and the necessary precursor synthesis genes (e.g., UGE) into the host genome.
• Metabolic Flux Balancing: Optimize gene expression levels of UDP-GlcNAc and UDP-GlcA pathway enzymes (e.g., GNA1, GlmS) using promoter libraries to achieve stoichiometric precursor balance .
• Fermentation Optimization: Optimize media and feeding strategies to manage the high viscosity, aiming for maximal HA titer while maintaining sufficient oxygen transfer.
• Product Quality Validation: Measure the final HA titer (g/L) and molecular weight (Daltons) using HPLC and SEC-MALLS (Size Exclusion Chromatography with Multi-Angle Laser Light Scattering).

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

Explore the potential for a safe, high-quality, and high-yield HA supply. CD Biosynsis provides customized strain and process engineering solutions:

• Detailed Hyaluronidase Knockout Verification Report and MW Stability Data.
• Consultation on bioreactor agitation and aeration strategies to overcome the high viscosity challenge at large scale.
• Experimental reports include complete raw data on HA titer, MW distribution and final product purity , essential for cosmetic and medical applications.