Astaxanthin Oxide Bioproduction Engineering Service

Astaxanthin Oxide (Asta-oxide or Astaxanthin epoxide) is a functional derivative of Astaxanthin, possessing potentially enhanced bioactivity in specific pharmaceutical and cosmetic applications. Traditional production routes face significant challenges: Multiple steps in chemical synthesis are required to introduce the epoxide moiety, resulting in low overall yield and the formation of numerous unwanted stereoisomers. Furthermore, the final product often exhibits low activity due to contamination with inactive isomers or inherent structural instability. Microbial biosynthesis offers a superior approach, leveraging enzymatic precision for specific and high-purity production.

CD Biosynsis offers a synthetic biology service focused on efficient Asta-oxide production using Saccharomyces cerevisiae (baker's yeast). Our core strategy involves modification of astaxanthin synthesis pathway in Saccharomyces cerevisiae . Astaxanthin (Asta) production in yeast requires heterologous expression of a carotenoid pathway (e.g., CrtE, B, I, Y, and P/CrtZ) to convert the precursor Isopentenyl Pyrophosphate (IPP) into Asta. We maximize the Asta precursor yield by optimizing gene expression and relieving flux bottlenecks in the MVA pathway. This is coupled with directed evolution of oxidase . We introduce a specific Monooxygenase (e.g., CYP450 or an Epoxidase) capable of selectively converting Astaxanthin to the desired Asta-oxide isomer. This Oxidase is optimized via directed evolution (e.g., saturation mutagenesis and screening) to achieve high catalytic activity and strict regioselectivity, ensuring the production of the most bioactive isomer while minimizing side products. This integrated approach simplifies the process, dramatically reduces inactive isomer formation, and ultimately yields a more potent and stable Asta-oxide product.

Pain Points

Developing a high-quality Asta-oxide product faces these key challenges:

• Multiple Steps in Chemical Synthesis: Chemical epoxidation is inefficient, requiring complex protection and deprotection steps and yielding a mixture of Asta cis/trans isomers and various mono- and di-epoxides, leading to low overall yield.
• Low Activity: Contamination with inactive epoxide stereoisomers from non-selective chemical processes reduces the overall bioactivity of the final commercial product.
• Astaxanthin Precursor Bottleneck: The heterologous Astaxanthin pathway in yeast is complex and requires the metabolic balancing of several introduced genes (Crt genes) to reach a high Asta concentration.
• Oxidase Selectivity: The final Oxidase must be precisely tailored to convert Astaxanthin to the Asta-oxide without over-oxidation or non-specific reactions, a challenge for naturally occurring enzymes.

A successful solution must ensure a high yield of the Astaxanthin precursor and establish a highly selective enzymatic epoxidation step.

Solutions

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

Modification of Astaxanthin Synthesis Pathway in S. cerevisiae

           

We enhance the MVA pathway (e.g., overexpression of tHMG1 ) and optimize the Crt gene cluster to maximize the concentration of the Astaxanthin precursor.

Directed Evolution of Oxidase

We use mutagenesis and high-throughput screening to evolve a Monooxygenase variant with enhanced activity and strict regioselectivity for the Astaxanthin-to-Asta-oxide conversion.

Competing Pathway Blockade

We delete or downregulate pathways that divert carbon flux from IPP (e.g., Ergosterol synthesis) to improve overall Astaxanthin yield.

Oxidative Stability Enhancement

We co-express antioxidant defense enzymes (e.g., Superoxide Dismutase) to protect the final Asta-oxide product from degradation within the cell.

This systematic approach overcomes metabolic bottlenecks and ensures the final enzymatic conversion is highly selective and efficient.

Advantages

Our Asta-oxide engineering service is dedicated to pursuing the following production goals:

High Bioactivity and Purity

Enzymatic conversion ensures production of the desired Asta-oxide isomer , solving the low activity issue from chemical synthesis.

Simplified and Cost-Effective Process

Eliminating multiple chemical steps and complex separation reduces manufacturing time and cost . [Image of Cost Reduction Icon]

Sustainable Production Route Icon

Yeast fermentation uses renewable resources, offering an eco-friendly alternative to resource-intensive chemical methods.

High Volumetric Productivity

The engineered yeast strain is capable of achieving high product titer in scalable fermentation platforms.

Food-Grade GRAS Host Icon

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

We provide a competitive, high-purity, and highly active biosynthetic route for Asta-oxide.

Process

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

• Astaxanthin Pathway Construction: Introduce and balance the heterologous Crt gene cluster into S. cerevisiae and upregulate the native MVA pathway.
• Oxidase Directed Evolution: Identify candidate Astaxanthin Epoxidases. Use random or site-directed mutagenesis followed by screening to evolve a variant with high Asta to Asta-oxide conversion specificity .
• Pathway Fine-Tuning: Optimize the relative expression levels of all Crt genes and the evolved Oxidase to maximize final Asta-oxide accumulation.
• Cofactor and Protection: Optimize the supply of essential NADPH cofactor and co-express antioxidant enzymes to prevent product degradation .
• Functional and Titer Assays: Validate the engineered strain in fed-batch culture, measuring the final Asta-oxide titer, yield, and purity (isomer ratio).
• Result Report Output: Compile a detailed Experimental Report including gene modification data, enzyme characterization, and fermentation metrics (volumetric titer and conversion rate) , supporting industrial scale-up.

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

Explore the potential for a stable, high-activity Asta-oxide supply. CD Biosynsis provides customized strain and enzyme engineering solutions:

• Detailed Epoxide Specificity and Titer Report , demonstrating the single-step conversion efficiency and desired isomer ratio.
• Consultation on optimized cell disruption and extraction protocols tailored for carotenoid derivative recovery from yeast biomass.
• Experimental reports include complete raw data on total Asta-oxide production (mg/L) and final product stability analysis , essential for pharmaceutical and cosmetic quality control.