Isoprene (Bio-Rubber Monomer) Metabolic Engineering Service

Isoprene (C5H8) is the key monomer used to produce synthetic rubber and elastomers . Conventional petrochemical synthesis is highly energy-intensive and non-renewable . Bio-based production faces challenges due to low carbon flux into the MEP or MVA Isoprenoid precursor pathways (IPP/DMAPP) . Furthermore, Isoprene is a highly volatile product that requires specialized collection and fermentation design to prevent loss and cell inhibition at high concentrations in the liquid phase (product inhibition)

CD Biosynsis offers a comprehensive metabolic engineering solution to achieve high-titer, efficient Isoprene production. Our strategy includes: Pathway Engineering: Overexpress all MEP pathway enzymes and the final Isoprene Synthase (IspS) . This removes bottlenecks in the key precursor pathway (MEP is often preferred in E. coli hosts). We also focus on Cofactor Optimization: Balance NADPH supply, a key cofactor for the MEP pathway . Multiple MEP steps are NADPH-dependent reductions (e.g., IspC, IspE). Finally, we address downstream recovery with Bioreactor Design: Implement in situ product stripping (e.g., gas-stripping bioreactors) to recover volatile Isoprene and reduce product inhibition .

Pain Points

Industrial Isoprene production via biosynthesis faces these key challenges:

• MEP/MVA Pathway Bottlenecks: The native isoprenoid precursor pathways (MEP in bacteria, MVA in yeast) are highly regulated and metabolically taxing, leading to low flux of IPP and DMAPP (the direct precursors to Isoprene Synthase).
• Cofactor Imbalance: The MEP pathway in E. coli requires a high supply of NADPH for reduction steps. If NADPH supply is not optimized, the pathway stalls and intermediates accumulate.
• Product Volatilization and Inhibition: Isoprene is highly volatile, leading to loss if not captured. Moreover, Isoprene is a non-polar solvent that is toxic to cells at high liquid concentrations, causing membrane damage and reduced cell viability .
• Competing Terpenoid Production: Native or synthetic pathways may divert IPP/DMAPP to other terpenoids (e.g., Farnesyl Pyrophosphate (FPP)), decreasing Isoprene yield .

A successful solution must maximize precursor flux and integrate an efficient in situ product recovery system.

Solutions

CD Biosynsis utilizes advanced metabolic and bioreactor engineering to optimize Isoprene production:

MEP Pathway Flux Optimization

           

We overexpress and balance all seven MEP enzymes (e.g., IspC, IspD) to increase the supply of IPP/DMAPP, the direct precursors.

Cofactor Engineering (NADPH)

We engineer the host (e.g., E. coli glycolysis) to favor NADPH regeneration (e.g., overexpressing G6PDH in PPP) to support MEP reduction steps and remove bottlenecks.

In Situ Product Stripping

We design a fermentation system using gas-stripping (air or inert gas flow) to continuously remove volatile Isoprene from the broth, mitigating toxicity and increasing productivity.

Block Competing Terpenoid Pathways

We delete genes (e.g., ispA, idi mutants) that divert IPP/DMAPP to longer-chain terpenoids (e.g., FPP or GGPP), maximizing flux to Isoprene Synthase (IspS).