3-Hydroxypropionic Acid (3-HP) High-Yield Biosynthesis Service

3-Hydroxypropionic Acid (3-HP) is a crucial bio-based platform chemical for producing various high-value downstream products, including acrylic acid, 1,3-propanediol, and biodegradable plastics. However, conventional biological production methods face significant challenges: low yield of traditional fermentation and substrate conversion rates often falling below 50% , limiting industrial viability.

CD Biosynsis offers a comprehensive metabolic engineering approach to establish a highly efficient 3-HP biosynthesis pathway. Our strategy focuses on the reconstruction of the glyoxylate cycle in Escherichia coli to optimize carbon flow and the overexpression of key enzymes in the beta-alanine pathway to boost the final conversion step. This dual approach maximizes both carbon efficiency and pathway activity, delivering a high-yield, high-conversion E. coli strain for industrial 3-HP production.

Pain Points

Achieving cost-effective, high-titer production of 3-HP via biological routes is constrained by several factors:

• Low Substrate Conversion Efficiency: The primary pathway from glucose often involves multiple steps with energy-intensive intermediates, resulting in substrate conversion rates well below the theoretical maximum (less than 50%) .
• Carbon Flux Bottlenecks: In native hosts, the carbon flow from central metabolism is often bottlenecked (e.g., at the glyoxylate shunt) or diverted to undesirable byproducts (like acetate), leading to low final 3-HP yield .
• Enzyme Activity and Stability: Key enzymes, particularly those responsible for the final conversion (e.g., the aldehyde dehydrogenase), often exhibit low specific activity or poor stability under high-concentration fermentation conditions.
• Product Toxicity and Inhibition: High concentrations of 3-HP can be toxic to the microbial host, leading to growth inhibition and low final titer .

A successful solution must reroute central carbon metabolism to maximize precursor availability and enhance the efficiency of the target pathway enzymes.

Solutions

CD Biosynsis utilizes advanced synthetic biology techniques to overcome 3-HP production limits:

Reconstruction of the Glyoxylate Cycle

           

We reconstruct or enhance the glyoxylate cycle in E. coli to provide an efficient anaplerotic route, bypassing the carbon-losing steps of the TCA cycle and maximizing the supply of the precursor succinate/oxaloacetate.

Overexpression of Key Enzymes in the β-Alanine Pathway

Key enzymes in the beta-alanine pathway (e.g., beta-alanine-pyruvate aminotransferase and 3-HP dehydrogenase) are overexpressed and balanced to ensure rapid and efficient conversion of the beta-alanine intermediate to the final 3-HP product.

Competing Pathway Knockout/Downregulation

Unwanted side pathways (e.g., acetate formation) and native enzymes that consume precursors are knocked out or downregulated to direct maximal carbon flux into the engineered 3-HP pathway.

Cofactor ($\text{NADH}/\text{NADPH}$) Optimization

Since 3-HP synthesis often involves redox reactions, we engineer the host to optimize the availability and regeneration of essential cofactors ($\text{NADH}$ or $\text{NADPH}$), crucial for high-rate conversion.

This systematic approach guarantees high yield and efficiency for scalable bioproduction of 3-HP.

Advantages

Partnering with CD Biosynsis for 3-HP strain development provides the following core benefits:

High Substrate Conversion Rate

Glyoxylate cycle reconstruction significantly improves carbon utilization, enabling substrate conversion rates close to the theoretical maximum.

Enhanced Carbon Flux

Strategic pathway engineering directs over 90% of the carbon source specifically toward 3-HP production, maximizing yield.

Robust Host Strain

The E. coli host is engineered for high tolerance to elevated 3-HP concentrations, supporting high-titer fed-batch fermentation.

Simplified Purification

Minimized byproduct formation due to pathway knockout leads to a cleaner fermentation broth, reducing downstream processing costs.

Efficient Enzyme Performance

Protein engineering optimizes key pathway enzymes, ensuring high specific activity and thermal stability for industrial scale-up.

We deliver strains ready for immediate integration into your large-scale biomanufacturing pipeline.

Process

CD Biosynsis's 3-HP strain engineering service follows a rigorous, multi-step workflow:

• Metabolic Bottleneck Identification: Perform Flux Balance Analysis (FBA) and gene expression profiling to identify carbon sinks and rate-limiting steps within the native E. coli metabolism.
• Glyoxylate Cycle Engineering: Introduce or upregulate key enzymes of the glyoxylate cycle (e.g., isocitrate lyase) to maximize C4 precursor supply to the downstream pathway.
• beta-Alanine Pathway Construction: Design and clone the beta-alanine pathway, using promoter engineering and codon optimization to balance the expression levels of all pathway enzymes.
• Cofactor and Redox Balance Optimization: Introduce or modify pathways (e.g., transhydrogenase) to ensure optimal NADPH or NADH regeneration required by 3-HP synthesis.
• High-Density Fermentation Validation: Test the final engineered strain in high-cell-density fed-batch fermentation to measure final titer, yield, and substrate conversion rate .
• Result Documentation and Transfer: Provide a comprehensive report including genetic maps, enzyme kinetics, fermentation performance data, and detailed protocols for industrial adoption.

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

Accelerate your entry into the bio-based chemical market. CD Biosynsis provides customized strain engineering solutions:

• Detailed Yield and Substrate Conversion Analysis Report , demonstrating the improved carbon efficiency.
• Consultation on optimizing the fermentation feeding strategy to mitigate product toxicity.
• Experimental reports include complete raw data on titer, yield, and strain stability , essential for regulatory compliance and scale-up planning.