1,3-Butanediol Biosynthesis via Engineered E. coli Service

1,3-Butanediol (1,3-BDO) is a high-demand commodity used extensively as a humectant in cosmetics and as a chiral intermediate in medicine. Its production faces dual challenges: petrochemical synthesis relies heavily on finite fossil resources , and early attempts at biosynthesis suffer from a low substrate conversion rate and poor yield from renewable feedstocks. This demands a robust, high-efficiency bio-based alternative.

CD Biosynsis offers a cutting-edge metabolic engineering solution focused on using renewable glycerol feedstock. Our core strategy involves the reconstruction of the glycerol metabolism pathway in Escherichia coli to maximize carbon flux towards the 1,3-BDO route. This is paired with the synergistic optimization of dehydrogenases and reductases —the key enzymes in the 1,3-BDO pathway—to enhance conversion efficiency and control the essential redox balance. We provide a genetically stable, high-titer E. coli strain that ensures reliable, eco-friendly production of 1,3-BDO.

Pain Points

Achieving industrial titers of bio-based 1,3-BDO is currently limited by:

• Low Substrate Conversion Rate: In early bioprocesses, the flux from the renewable substrate (often glycerol) into the 1,3-BDO pathway is inefficient, resulting in a low overall conversion rate and high residual substrate concentration.
• Redox Imbalance: The synthetic pathway for 1,3-BDO involves multiple steps that require a precise supply of NAD(P)H/NAD(P)+ cofactors. An imbalance often leads to a stagnant reaction or the accumulation of unwanted side products (e.g., 1,2-propanediol).
• Competition from Native Pathways: Carbon flow is easily diverted to the host's native metabolism for biomass growth and the production of undesired byproducts (acetate, ethanol), significantly reducing the final 1,3-BDO yield .
• Enzyme Sub-Optimality: The heterologous dehydrogenases and reductases required for the 1,3-BDO pathway often lack sufficient specific activity or substrate affinity for industrial-scale efficiency.

A successful solution must establish high-efficiency carbon flow from glycerol and tightly control the required redox reactions.

Solutions

CD Biosynsis employs a holistic strategy of metabolic and enzyme engineering to achieve high-titer, bio-based 1,3-BDO:

Reconstruction of Glycerol Metabolism Pathway

           

We delete competing native pathways (acetate, ethanol production) and upregulate key enzymes in the central metabolic path (e.g., EMP pathway) to ensure maximum flux from glycerol directly towards the 1,3-BDO intermediate (acetyl-CoA).

Synergistic Optimization of Dehydrogenases and Reductases

The entire cascade of dehydrogenases and reductases is rationally engineered and co-expressed. This ensures a balanced demand and supply of NAD(P)H cofactors, leading to rapid, high-yield conversion of the intermediate to the final 1,3-BDO product.

Cofactor Regeneration System Engineering

We specifically engineer the host's central metabolism to boost the intracellular NAD(P)H supply . This is critical for driving the multiple reduction steps in the 1,3-BDO pathway, overcoming the major redox bottleneck.

Pathway Balancing and Flux Control

We use synthetic biology tools to finely tune the expression levels of all heterologous enzymes, ensuring a perfectly balanced metabolic flux and minimizing the accumulation of toxic or undesirable intermediates.

This integrated approach establishes a dedicated, high-flux pathway for sustainable 1,3-BDO production from low-cost renewable glycerol.

Advantages

Choosing CD Biosynsis's 1,3-BDO engineering service offers the following core value:

Sustainable, Bio-Based Production

Utilizes renewable glycerol feedstock (a biodiesel byproduct), eliminating reliance on volatile fossil resources for a greener product life cycle.

High Substrate Conversion Rate

Pathway reconstruction and enzyme optimization ensure maximum flux, leading to a significantly higher conversion of glycerol to 1,3-BDO .

Enhanced Redox Control

Synergistic enzyme optimization overcomes the NAD(P)H bottleneck, ensuring the reduction reactions proceed rapidly for maximum yield .

High Purity for Cosmetic Grade

The engineered pathway produces 1,3-BDO with minimal byproduct formation , simplifying downstream purification for high-grade applications.

Utilizes Low-Cost Glycerol Feedstock

The reliance on crude glycerol (a low-value byproduct) as a primary carbon source drastically reduces raw material costs.

We deliver the technology to transition 1,3-Butanediol production to a robust, sustainable, and economically competitive bio-process.

Process

CD Biosynsis's 1,3-BDO engineering service follows a standardized research workflow, ensuring every step is precise and controllable:

• Metabolic Analysis and Target Definition: Define the target 1,3-BDO titer and conversion rate. Conduct a Flux Balance Analysis (FBA) of the glycerol pathway to identify flux leakage and NAD(P)H bottlenecks.
• Pathway Reconstruction and Competition Deletion: Use CRISPR-Cas to delete major byproduct pathways (acetate, ethanol) and integrate the 1,3-BDO pathway genes into the E. coli chromosome.
• Enzyme and Cofactor Optimization: Rational design and screening to identify and co-express the most efficient dehydrogenase/reductase combinations . Engineer native pathways to boost NAD(P)H regeneration .
• Pathway Balancing and Integration: Systematically tune the expression of all pathway genes to achieve optimal flux balance and minimize intermediate accumulation, ensuring high-efficiency final conversion.
• Performance Validation Experiments: Conduct comparative fed-batch fermentation experiments using glycerol as the sole carbon source, measuring the final 1,3-BDO titer, substrate conversion rate, and byproduct formation using HPLC/GC-MS.
• Result Report Output: Compile a Strain Engineering Experimental Report that includes genetic maps, FBA data, fermentation kinetics, and a final titer/conversion certificate, supporting industrial scale-up.

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

Transition to sustainable, high-performance 1,3-Butanediol production! CD Biosynsis provides customized strain engineering solutions:

• Detailed Glycerol Pathway Flux and Redox Balance Report , guiding optimal feeding and process control.
• Contracted clients receive consultation on optimizing the fermentation oxygen supply to facilitate required redox steps.
• Experimental reports include complete raw data on titer, conversion rate, and byproduct purity , essential for market entry and technical claims.