Farnesene Engineering Service

Farnesene is a high-value bio-monomer precursor for Bio-monomers Coatings/Jet Fuel e.g. lubricants, advanced polymers, and jet fuel blending. Microbial production faces key challenges: Low concentration and high cost of Farnesene from natural plant oils make sourcing difficult. In yeast hosts, the major metabolic hurdle is competition for the precursor FPP Farnesyl Diphosphate with native sterol synthesis pathways e.g. Ergosterol synthesis. This limits the yield and titer achievable.

CD Biosynsis offers a tailored engineering strategy to maximize Farnesene production in yeast: Pathway Engineering: Overexpress all key enzymes in the MVA pathway to enhance FPP supply . This ensures a high flux into the terpenoid production pathway. To address precursor competition, we perform Knockout ERG9 Squalene Synthase to eliminate the major competitive sink for FPP . ERG9 consumes FPP to produce sterols, so its deletion redirects FPP flow toward Farnesene. Finally, we ensure efficient product formation by employing Introduce Farnesene Synthase FAS for final product synthesis , using a highly active exogenous enzyme.

Pain Points

The industrial bioproduction of Farnesene presents these main difficulties:

• Precursor Competition: Farnesyl Diphosphate FPP is a precursor for both the target product Farnesene and essential native compounds like Ergosterol in yeast. The native pathway acts as a major metabolic sink, drastically reducing target yield.
• Low Pathway Flux: The native Mevalonate MVA pathway that generates FPP is tightly regulated in yeast, resulting in low overall carbon flux toward the terpenoid backbone.
• Product Toxicity and Volatility: Farnesene is a volatile organic compound that can be toxic to the host cell at high concentrations, complicating high-titer fermentation and recovery.
• Expensive Downstream Processing: Achieving high purity for jet fuel or polymer applications requires efficient separation of the low-concentration product from the fermentation broth, contributing to high costs.

A cost-effective strategy must maximize the FPP precursor pool and redirect its flow exclusively to Farnesene.

Solutions

CD Biosynsis utilizes advanced pathway and host engineering to maximize Farnesene production:

MVA Pathway Engineering

           

We overexpress key rate-limiting enzymes e.g. HMG-CoA reductase, HMG-CoA synthase in the MVA pathway to significantly increase the pool of FPP precursors available for Farnesene synthesis.

Competitive Sink Knockout

We use CRISPR-Cas9 or similar tools to knockout the ERG9 Squalene Synthase gene , effectively eliminating the major competitive sink for FPP and redirecting carbon flux to the target product.

Exogenous FAS Introduction

We introduce a highly active, stable Farnesene Synthase FAS gene from a heterologous source to catalyze the final step, converting FPP into the volatile Farnesene product.

Two-Phase Fermentation System

We implement a two-phase fermentation system using an organic solvent to continuously extract the volatile product, reducing product toxicity and enhancing final titer . [Image of High Conversion Efficiency Icon]

This integrated approach maximizes FPP precursor availability and minimizes competitive loss, leading to high-titer Farnesene production.

Advantages

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

Maximize FPP Precursor Pool Icon

MVA pathway engineering ensures a high flux of carbon into the terpenoid synthesis route, increasing precursor FPP availability.

Eliminate Competitive Byproducts Icon

ERG9 knockout redirects nearly all FPP from native sterol synthesis to the target Farnesene product. [Image of Cost Reduction Icon]

High Titer and Reduced Toxicity Icon

The two-phase fermentation system allows for high product accumulation while mitigating cell toxicity.

Scalability and Economic Viability Icon

Optimized strain and two-phase separation support industrial scale-up for high-volume bio-monomer production.

High Conversion Efficiency Icon

Engineered pathway and targeted knockout lead to minimal carbon loss to byproducts, improving overall yield.

We deliver an economically optimized yeast platform for high-titer Farnesene biosynthesis.

Process

Our Farnesene strain engineering service follows a rigorous, multi-stage research workflow:

• Gene Cloning and MVA Pathway Overexpression: Construct a multi-gene expression cassette containing all key MVA pathway enzymes and the Farnesene Synthase FAS gene.
• Targeted Knockout: Perform ERG9 gene deletion using CRISPR-Cas9 or homologous recombination to block the sterol synthesis pathway and redirect FPP flux.
• Host Optimization: Integrate the optimized pathway into the yeast genome and optimize growth conditions for maximal precursor flow e.g. medium composition, temperature, aeration.
• Fermentation Process Development: Develop a two-phase fermentation protocol using a suitable solvent or resin for in-situ product extraction, maximizing titer and reducing cell stress.
• Product Titer and Purity Analysis: Quantify the final Farnesene titer g/L and purity via GC-MS and validate the stability of the engineered strain over repeated fermentation cycles.

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

Explore the potential for a high-titer, cost-effective Farnesene supply. CD Biosynsis provides customized strain and process engineering solutions:

• Detailed Farnesene Titer Report g/L from optimized fermentation runs using two-phase systems.
• Consultation on downstream processing and product separation protocols for jet fuel or polymer grade purity.
• Experimental reports include complete raw data on strain stability, gene expression levels, and precursor FPP pool measurements .