Squalane Bioproduction Engineering Service

Squalane is a saturated hydrocarbon widely prized in the cosmetics and pharmaceutical industries for its excellent emollient, moisturizing, and skin penetration properties. Historically, the precursor, Squalene, was sourced from shark liver oil, a practice that shark extraction damages ecology and is ethically untenable. While chemical synthesis and plant extraction are alternatives, they suffer from high cost for chemical synthesis and yield low concentration. Biosynthesis offers a scalable, eco-friendly, and cost-effective alternative by producing Squalene from simple sugars, followed by enzymatic conversion to Squalane.

CD Biosynsis offers a synthetic biology service focused on high-yield Squalane production using Saccharomyces cerevisiae (baker's yeast). Our core strategy involves modification of squalene synthesis pathway in Saccharomyces cerevisiae . Squalene is naturally synthesized via the highly regulated Mevalonate (MVA) pathway. We maximize Squalene accumulation by overexpressing key MVA pathway enzymes (e.g., tHMG1) and Squalene Synthase (ERG9) while deleting or downregulating pathways that consume Squalene (e.g., conversion to Ergosterol). This is coupled with heterologous expression of hydrogenase . Squalane is derived by the saturation (hydrogenation) of Squalene's six double bonds. We introduce a heterologous, NADPH-dependent Hydrogenase (e.g., from a specific microbial source) into the engineered yeast strain. This enzyme catalyzes the conversion of Squalene to the stable, fully saturated Squalane product in a single, clean enzymatic step. This integrated approach bypasses destructive animal sourcing and the multi-step, high-energy requirements of chemical hydrogenation, delivering a sustainable, high-purity Squalane.

Pain Points

Sustainable and cost-effective Squalane production faces these key challenges:

• Shark Extraction Damages Ecology: Sourcing from shark liver oil is unsustainable and harms marine biodiversity , leading to ethical and supply chain risks.
• High Cost for Chemical Synthesis: Chemical hydrogenation of Squalene requires high pressure, expensive noble metal catalysts (e.g., Pd/C), and large energy input , resulting in high operational costs.
• Low Squalene Precursor Yield: In yeast, Squalene is an intermediate that is rapidly converted to Ergosterol. Tightly regulated flux and competing pathways limit the Squalene titer.
• Product Instability: The biosynthetic product, Squalene, contains six double bonds and is prone to auto-oxidation and degradation upon exposure to air, making handling difficult and reducing shelf life.

A successful solution must shift the production away from animal sources, maximize Squalene yield, and enzymatically stabilize the final product.

Solutions

CD Biosynsis utilizes advanced metabolic and enzyme engineering to optimize Squalane production in S. cerevisiae:

Modification of Squalene Synthesis Pathway in S. cerevisiae

           

We overexpress rate-limiting MVA enzymes (tHMG1) and Squalene Synthase (ERG9), while deleting Squalene Epoxidase (ERG1) , the enzyme that consumes Squalene to produce Ergosterol.

Heterologous Expression of Hydrogenase

We introduce a specific NADPH-dependent Hydrogenase that catalyzes the Squalene-to-Squalane conversion, offering a clean, metal-free hydrogenation step.

NADPH Cofactor Engineering

We co-express enzymes (e.g., G6PDH) to regenerate NADPH , the essential cofactor for the Hydrogenase enzyme, ensuring sustained catalytic activity.

Lipid Body Morphology Optimization

We engineer the strain to increase the size and capacity of lipid droplets (storage organelles), maximizing the storage capacity for the hydrophobic Squalane product.

This systematic approach ensures high Squalene precursor yield and its subsequent clean, enzymatic stabilization into Squalane.

Advantages

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

Eco-Friendly and Ethical Sourcing

Production from yeast completely eliminates reliance on shark liver oil , supporting marine conservation and ethical practices.

Clean, Metal-Free Hydrogenation

Enzymatic conversion avoids the use of expensive, high-pressure equipment and noble metal catalysts , lowering capital and operational costs.

High Product Stability

The final saturated Squalane is highly stable and resistant to oxidation , unlike the Squalene intermediate, ensuring long shelf life.

High Fermentation Titer

Metabolic engineering ensures a maximal diversion of carbon from sugar to the Squalene pathway, leading to high accumulation.

Food-Grade GRAS Host Icon

Using S. cerevisiae ensures the final product is compatible with cosmetic and health supplement regulatory standards .

We provide a sustainable and economically competitive biosynthetic route for high-quality Squalane.

Process

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

• Squalene Precursor Strain Construction: Overexpress tHMG1 and ERG9 and delete ERG1 (the Squalene Epoxidase gene) to maximize Squalene accumulation.
• Hydrogenase Expression: Identify and heterologously express a highly efficient, NADPH-dependent microbial Hydrogenase specific for Squalene saturation.
• Cofactor Optimization: Co-express NADPH regeneration enzymes (e.g., G6PDH) to ensure sufficient reducing power for the Hydrogenase reaction.
• Toxicity and Storage Optimization: Engineer the yeast membrane and lipid body composition to increase Squalane storage capacity and improve cell tolerance to the high-concentration product.
• Fermentation Performance Validation: Test the final engineered strain in fed-batch fermentation to assess Squalane titer and final product purity (ratio of Squalane to Squalene).
• Result Report Output: Compile a detailed Experimental Report including gene modification data, enzyme characterization, and fermentation metrics (volumetric titer and conversion efficiency) , supporting industrial scale-up.

Technical communication is maintained throughout the process, focusing on timely feedback regarding yield and saturation efficiency.

Explore the potential for a stable, eco-friendly Squalane supply. CD Biosynsis provides customized strain and enzyme engineering solutions:

• Detailed Squalane Titer and Saturation Efficiency Report , demonstrating the complete conversion of Squalene to Squalane.
• Consultation on optimized cell lysis and extraction protocols tailored for lipid body-stored Squalane recovery.
• Experimental reports include complete raw data on final Squalane yield (g/L) and fatty acid analysis , essential for quality control in cosmetic applications.