Phytosphingosine Bioproduction Engineering Service

Phytosphingosine (PS) is a naturally occurring long-chain sphingoid base with potent antimicrobial and anti-acne properties , making it a key component in advanced cosmetics and skincare, often used as a direct precursor for Ceramide production. Industrial production methods struggle with efficiency. Complex purification and low yield from traditional yeast extraction result in high manufacturing costs. Furthermore, Chemical synthesis is environmentally burdensome , requiring toxic reagents and leading to mixtures of less active stereoisomers. The biosynthetic approach focuses on enhancing the native yeast pathway, which is often limited by the activity of rate-limiting enzymes responsible for long-chain sphingolipid synthesis.

CD Biosynsis offers a comprehensive metabolic engineering solution, utilizing Saccharomyces cerevisiae (yeast) as the preferred host due to its native sphingolipid pathway. To significantly boost the final product titer, we Overexpress genes responsible for long-chain sphingolipid synthesis in yeast, such as Serine Palmitoyltransferase (Lcb 1/Lcb2) . SPT is the first and rate-limiting enzyme in the sphingoid base pathway; its overexpression drastically increases the flux from Serine and Palmitoyl-CoA to Phytosphingosine. In addition, we Optimize fermentation conditions and downstream processes to improve the efficiency of the in vitro synthesis and purification of non-natural long-chain sphingoid bases . This includes minimizing competing pathways and improving the desaturation (Des1) and hydroxylation (Sur2) steps needed to convert Sphinganine to the final Phytosphingosine structure, ensuring high yield and high stereoconfigurational purity.

Pain Points

Industrial Phytosphingosine production faces these key challenges:

• Complex Purification and Low Yield from Yeast Extraction: PS exists as part of complex sphingolipids within the yeast membrane. Isolation requires harsh hydrolysis and extensive chromatography, leading to low yield and high purification costs .
• Stereochemical Purity Issues with Chemical Synthesis: Chemical routes are multi-step and often yield racemic mixtures of sphingoid bases that lack the bioactive natural stereoconfiguration , and the use of toxic reagents is environmentally burdensome.
• Limiting Sphingoid Base Synthesis: The rate-limiting enzyme in sphingoid base formation, Serine Palmitoyltransferase (SPT complex Lcb 1/Lcb2) , is tightly regulated in native yeast, leading to an Insufficient precursor pool for high PS production.
• Pathway Competition and Product Diversion: In yeast, PS is quickly converted into complex Ceramides and Glycosphingolipids (e.g., IPCs), reducing the final yield of the desired free Phytosphingosine product.

A successful solution must ensure a massive supply of the sphingoid base precursor and prevent its diversion into complex lipids.

Solutions

CD Biosynsis utilizes advanced metabolic engineering to optimize Phytosphingosine production in yeast:

Overexpress Key Enzymes (Lcb 1/Lcb2) for Precursor Boost

           

We overexpress the Lcb 1/Lcb2 (SPT) complex along with cofactors (Tsc3) to significantly increase the flux into the sphingoid base pathway.

Block Downstream Pathways to Prevent Product Diversion

We use gene knockouts (e.g., Lac 1/Lag1 Ceramide Synthases) to stop the conversion of PS to Ceramides , maximizing the free Phytosphingosine yield.

Optimize Fermentation and In Vitro Processes

We tune the growth environment and downstream hydrolysis protocols to maximize free PS recovery and purity of the bioactive isomer.

Fine-Tune Desaturation and Hydroxylation

We optimize the activity of Sur2 and Des1 genes to efficiently convert Sphinganine to Phytosphingosine with high specificity.

This integrated approach significantly increases the initial production rate and simplifies the final Phytosphingosine recovery.

Advantages

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

High Titer and Yield

SPT overexpression and pathway blocking dramatically increases the initial PS pool size, resulting in higher final recoverable yield.

High Bioactive Purity Icon

Enzymatic biosynthesis in yeast ensures the exclusive production of the D-erythro isomer , which is the highly active natural form, avoiding racemic mixtures.

Reduced Environmental Burden Icon

Microbial fermentation replaces multi-step chemical synthesis, eliminating the use of toxic reagents. [Image of Cost Reduction Icon]

Control Over Chain Length Icon

The system can be tuned to produce specific long-chain bases (C18, C20) by optimizing the fatty acid precursor supply.

Consistent Supply Chain Icon

Production is independent of seasonal variability and can be scaled to meet industrial demand reliably.

We provide a reliable, high-purity, and high-titer source for bioactive Phytosphingosine.

Process

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

• Precursor Synthesis Boosting: Overexpress the key enzymes Lcb 1/Lcb2 (SPT) to maximize the flux from Serine to Sphinganine.
• Terminal Synthesis Optimization: Engineer and optimize the hydroxylation (Sur2) and desaturation (Des1) steps to efficiently convert Sphinganine to Phytosphingosine.
• Product Diversion Blockade: Knockout downstream Ceramide synthase genes (Lac 1/Lag1) to prevent Phytosphingosine acylation.
• Downstream Process Tuning: Optimize lipid extraction and hydrolysis protocols for maximum release and purification of free PS from the yeast cell.
• Titer and Purity Validation: Validate the engineered strain in fermentation, measuring the final Phytosphingosine titer (g/L) and stereoconfigurational purity (LC-MS).

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

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

• Detailed Sphinganine Precursor Concentration Report , demonstrating the effect of SPT overexpression.
• Consultation on optimized lipid hydrolysis and purification protocols for the final product.
• Experimental reports include complete raw data on Phytosphingosine titer, yield, and final purity metrics , essential for high-end cosmetic manufacturing.