Putrescine Engineering Service

Putrescine is a crucial diamine intermediate used primarily for synthesizing high-performance Bio-monomers (Nylon 4,4), a sustainable alternative to petrochemical-based nylons. Its production faces two main challenges: Chemical synthesis is petrochemical-based and toxic; fermentation yield is limited by Putrescine toxicity. The toxicity issue significantly caps the yield attainable through microbial fermentation.

CD Biosynsis employs advanced metabolic and host engineering to overcome these limitations: Metabolic Engineering: Engineer E. coli to overexpress Ornithine Decarboxylase (ODC) and Knockout Putrescine degradation pathways. This redirects cellular carbon flow for high production. Crucially, we enhance tolerance via Toxicity Tolerance: Modify cell membranes or introduce efflux systems. This allows the microbial host to withstand high concentrations of the product, resulting in significantly higher yields and a more economically viable biomanufacturing process, entirely moving away from toxic petrochemical routes.

Pain Points

The transition to bio-based Putrescine is hindered by these challenges:

• Petrochemical Reliance and Toxicity: Traditional synthesis uses toxic chemicals and is reliant on non-renewable petrochemicals, leading to environmental and safety concerns.
• Product Toxicity to Host: As a diamine, Putrescine is toxic to microbial hosts e.g. E. coli by disrupting cell membrane integrity, limiting final fermentation titer and yield.
• Low Conversion Efficiency: Native microbes possess pathways that degrade Putrescine or shunt precursors e.g. ornithine into other metabolites, reducing product selectivity.
• Expensive Downstream Processing: Low titer in fermentation necessitates large volumes of broth to be processed, driving up recovery costs for this water-soluble diamine.

High yield bio-production requires resolving Putrescine toxicity.

Solutions

CD Biosynsis implements a two-pronged strategy focusing on Metabolic Flux and Host Tolerability:

Ornithine Decarboxylase Overexpression

           

We overexpress key biosynthetic enzymes e.g. ODC to drive high flux from ornithine to Putrescine and knockout degradation pathways.

Membrane and Efflux System Modification

We engineer the cell membrane lipid composition or introduce efflux pump genes to increase host tolerance to high Putrescine titer.

Biosensor-Guided Pathway Tuning

We utilize Putrescine-responsive biosensors to dynamically control gene expression and fine-tune the rate of synthesis and efflux.

Customized Downstream Integration

We develop integrated fermentation and separation systems e.g. in situ product removal to mitigate toxicity and reduce processing cost.

These strategies result in a high-titer, bio-based, and sustainable Putrescine production platform.

Advantages

Our Putrescine engineering service offers these core benefits:

High Titer Bio-Based Yield

Combining flux optimization and tolerance enhancement breaks the toxicity-limited yield barrier, achieving commercially relevant Putrescine titer.

Sustainable Nylon Precursor

Production from renewable sugars or low-cost biomass replaces petrochemicals, providing a sustainable monomer for Nylon 4,4.

Improved Host Robustness

Engineered membrane or efflux systems create a more robust microbial host that performs reliably under high product concentrations and industrial conditions.

Simplified Purification

The higher titer achieved in fermentation reduces the volume of broth required for downstream processing, saving time and energy in diamine recovery.

Avoids Toxic Byproducts

The fermentation route replaces harsh chemical synthesis steps, eliminating the generation of toxic intermediates and waste.

We deliver a high-yield, clean, and sustainable Putrescine production system.

Process

Our Putrescine engineering service follows a rigorous metabolic and host engineering workflow:

• Biosynthetic Pathway Construction: Clone and overexpress Ornithine Decarboxylase ODC gene and optimize its codon usage for E. coli.
• Degradation Pathway Inactivation: Design and execute CRISPR/Cas9-mediated knockouts of Putrescine degradation and competing metabolic genes.
• Toxicity Tolerance Engineering: Introduce high-efficiency diamine efflux pump genes or modify membrane lipid synthesis for enhanced host robustness.
• Fermentation Optimization: Develop optimized fed-batch or chemostat protocols to sustain high Putrescine titer without compromising cell viability.
• Purity and Titer Validation: Utilize HPLC or GC-MS analysis to validate final Putrescine titer, yield, and purity for polymerization.

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

Explore the potential for a high-yield, bio-based Putrescine supply. CD Biosynsis provides customized strain and process engineering solutions:

• Detailed Putrescine Titer, Yield, and Conversion Efficiency Reports g/L, percent theoretical.
• Consultation on downstream Nylon 4,4 polymerization and diamine purification strategies using the high-titer broth.
• Experimental reports include complete raw data on metabolic flux analysis, cell tolerance studies, and efflux pump performance.