Biobased Isoprene Precursor Engineering Service

Isoprene is a foundational building block used in the production of synthetic rubber (polyisoprene), resins, and various specialty chemicals. Currently, its production is dominated by petrochemical synthesis that relies on crude oil , creating price volatility and environmental concerns. The alternative route, microbial biosynthesis, currently struggles with low yield , hindering its ability to compete economically with petroleum-based production.

CD Biosynsis offers a synthetic biology service focused on engineering the microbial host Escherichia coli . Our core strategy involves the modification of the Escherichia coli mevalonate pathway (MEP or MVA) , an engineered route for terpenoid synthesis, to maximize carbon flux toward the direct precursor (DMAPP). This is combined with the directed evolution of isoprene synthase to enhance its specific activity, stability, and catalytic turnover rate. This integrated approach aims to deliver a high-yield, cost-effective, and sustainable bioproduction route for this critical chemical monomer.

Pain Points

Achieving commercial competitiveness for biobased Isoprene requires overcoming these biological and economic hurdles:

• Petroleum Dependence: The industry is heavily reliant on fossil fuels , leading to environmental costs and susceptibility to crude oil price swings.
• Low Biosynthesis Yield: Microbial hosts often exhibit low yield of Isoprene because native metabolic pathways are complex, and carbon is diverted to competing growth pathways.
• Enzyme Activity Limitation: The activity of the key enzyme, isoprene synthase , may be inherently low or unstable within the engineered microbial environment, limiting the final product formation rate.
• Toxicity and Volatility: Isoprene is a volatile and mildly toxic compound, and high concentrations can inhibit host cell growth and productivity during fermentation.

A competitive solution must focus on maximizing the flux through the synthetic pathway and enhancing enzyme performance.

Solutions

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

Escherichia coli Mevalonate Pathway Modification

           

We employ genome editing to upregulate rate-limiting enzymes in the MVA pathway, ensuring a high intracellular concentration of the DMAPP precursor.

Directed Evolution of Isoprene Synthase

We use molecular evolution techniques to create synthase variants with enhanced catalytic efficiency (kcat) and improved stability within the E. coli cytoplasm, maximizing the conversion rate of DMAPP to Isoprene.

Competing Pathway Knockout

We delete genes that divert DMAPP to unwanted terpenoids (e.g., farnesyl diphosphate) or cellular growth products, thus focusing the carbon flux towards Isoprene.

Gas Stripping and Recovery Optimization

We design fermentation processes that allow for efficient in-situ removal of volatile Isoprene (gas stripping) to minimize cellular toxicity and improve product recovery.

This systematic approach is focused on maximizing the metabolic output of Isoprene and enhancing the key enzyme's performance.

Advantages

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

Sustainable Production Route

Using renewable carbon sources (sugars) replaces dependence on non-renewable crude oil , offering a green chemical manufacturing path.

Enhanced Bioconversion Efficiency

Pathway and enzyme engineering aim to achieve high molar yield of Isoprene from the supplied glucose or feedstock.

Improved Enzyme Kinetics

Directed evolution is focused on maximizing the catalytic turnover rate of isoprene synthase, increasing productivity.

Reduced Cellular Toxicity

Optimized fermentation combined with gas stripping aims to maintain cell viability at high production rates by removing the volatile product.

Cost Reduction Potential

High yield and low-cost microbial fermentation are focused on achieving cost competitiveness with petrochemical production. [Image of Cost Reduction Icon]

We provide a biosynthetic platform aimed at maximizing the yield and cost-effectiveness of biobased Isoprene production.

Process

Our Biobased Isoprene strain engineering service follows a standardized, iterative research workflow:

• Metabolic Pathway Construction: Integrate the heterologous MVA pathway into the E. coli genome or plasmid, ensuring balanced expression of all pathway enzymes.
• Rate-Limiting Step Optimization: Apply genome editing to upregulate key rate-limiting enzymes within the MVA pathway to maximize DMAPP supply.
• Isoprene Synthase Evolution: Generate mutant libraries of isoprene synthase and screen for variants with superior kcat and stability under fermentation conditions.
• Competing Pathway Silencing: Systematically delete or downregulate genes that consume DMAPP or divert carbon away from the MVA pathway.
• Fermentation and Recovery Validation: Test the final engineered strain in a specialized fermentation setup coupled with a gas stripping unit to measure volumetric productivity and recovery efficiency.
• Result Report Output: Compile a detailed Experimental Report including pathway modification data, enzyme kinetics, and fermentation metrics (yield, titer, and recovery efficiency) , supporting scale-up and economic analysis.

Technical communication is maintained throughout the process, focusing on timely feedback regarding Isoprene yield and enzyme performance.

Explore the potential for a sustainable, high-yield Isoprene supply. CD Biosynsis provides customized strain and enzyme engineering solutions:

• Detailed Metabolic Flux and Yield Analysis Report , illustrating the success of pathway modifications and precursor supply.
• Consultation on fermentation and gas recovery strategies optimized for the volatile Isoprene product.
• Experimental reports include complete raw data on enzyme kinetics, volumetric productivity, and final product purity , essential for industrial adoption.