Zeaxanthin Oxide Bioproduction Engineering Service

Zeaxanthin Oxide (e.g., Zeaxanthin Epoxide) is a carotenoid derivative with enhanced antioxidant and photoprotective properties, making it valuable in the pharmaceutical and cosmetic industries. Current production methods often involve chemical synthesis from natural Zeaxanthin , which suffers from Low efficiency in chemical synthesis due to multi-step reactions, low yield, and lack of stereoselectivity. Additionally, the Zeaxanthin Oxide product itself has poor stability due to sensitivity to light, heat, and oxygen, leading to rapid degradation during processing and storage.

CD Biosynsis offers a synthetic biology service focused on the clean and efficient production of specific Zeaxanthin Oxides using Escherichia coli (E. coli). Our core strategy addresses both efficiency and stability issues. The efficiency bottleneck ( low efficiency in chemical synthesis ) is overcome by Modification of zeaxanthin synthesis pathway in Escherichia coli . We engineer E. coli to maximize the metabolic flux towards the Zeaxanthin precursor, followed by the co-expression of the Zeaxanthin pathway genes (CrtE, B, I, Y, Z). We introduce a highly specific, rate-limiting Oxidase (e.g., cytochrome P}450$ or monooxygenase) responsible for the conversion of Zeaxanthin to its Oxide. The key innovation is the Directed evolution of oxidase . We apply directed evolution techniques (saturation mutagenesis, DNA shuffling) to this Oxidase to significantly boost its catalytic efficiency and stereoselectivity for the desired oxide isomer. The stability challenge ( poor stability ) is addressed by generating specific, more robust oxide isomers and potentially co-expressing antioxidant E. coli systems. This integrated biosynthetic route replaces inefficient chemical steps with a single, highly efficient enzymatic conversion, yielding a superior and purer product.

Pain Points

Developing a high-quality Zeaxanthin Oxide product faces these key challenges:

• Low Efficiency in Chemical Synthesis: Chemical epoxidation is non-selective, leading to a mixture of diastereomers (e.g., cis and trans epoxides) and low overall yield of the most active isomer, causing low efficiency .
• Poor Stability: Zeaxanthin Oxides, particularly epoxides, are thermodynamically unstable and readily isomerize or decompose under heat, light, and low pH conditions, resulting in poor stability during purification and formulation.
• Oxidase Selectivity: The newly introduced Oxidase must be optimized to exclusively convert Zeaxanthin into the desired Oxide isomer without forming unwanted side products or acting non-specifically on other carotenoids.
• Carotenoid Pathway Flux: Achieving a high concentration of the precursor Zeaxanthin in E. coli requires the coordination of multi-enzyme MEP and Crt pathways to avoid metabolic bottlenecks.

A successful solution must ensure robust Zeaxanthin production and introduce a highly active, specific Oxidase for conversion.

Solutions

CD Biosynsis utilizes advanced metabolic and enzyme engineering to optimize Zeaxanthin Oxide production in E. coli:

Modification of Zeaxanthin Synthesis Pathway in E. coli

           

We upregulate the central carbon flux to the MEP pathway and express the complete Crt gene cluster (CrtE, B, I, Y, Z) to maximize Zeaxanthin precursor titer.

Directed Evolution of Oxidase

We use mutagenesis and high-throughput screening to enhance the Oxidase's catalytic efficiency and stereoselectivity for the Zeaxanthin Oxide conversion, addressing low efficiency .

Protein Stabilization Engineering

We introduce stability-enhancing point mutations into the Oxidase and engineer the host environment to reduce reactive oxygen species (ROS), mitigating Oxide degradation ( poor stability ).

Subcellular Localization Optimization

We target the Oxidase to specific cellular membranes (e.g., inner membrane) to maximize its activity near the membrane-localized Zeaxanthin.

This integrated approach ensures high Zeaxanthin supply and a highly specific enzymatic conversion to the desired Oxide isomer with improved stability.

Advantages

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

High Specificity and Purity

The Directed Evolution of the Oxidase ensures the production of the single, most active Oxide isomer , solving the stereoselectivity problem of low efficiency chemical routes.

Enhanced Product Stability

Host engineering and specific isomer selection yield a more robust product, mitigating the issue of poor stability during processing.

Simplified Production Process Icon

Replacing complex chemical steps with a highly efficient microbial fermentation simplifies manufacturing and reduces reliance on harsh solvents.

High Titer and Yield Icon

Optimized MEP and Crt pathways ensure high precursor flow, maximizing the final Oxide yield per fermentation batch.

Reduced Environmental Impact Icon

Biosynthesis is a greener manufacturing route, avoiding the toxic byproducts and high energy consumption of chemical synthesis.

We provide a sustainable, high-performance, and high-purity biosynthetic route for Zeaxanthin Oxide.

Process

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

• Pathway Construction: Introduce and optimize the Zeaxanthin gene cluster (CrtE, B, I, Y, Z) and upregulate the native MEP pathway in E. coli.
• Oxidase Engineering: Identify, clone, and optimize the expression of the specific Oxidase for Zeaxanthin Oxide formation.
• Directed Evolution: Apply random and saturation mutagenesis to the Oxidase gene, followed by high-throughput screening for variants with enhanced specific activity and stereoselectivity .
• Host Stabilization: Engineer host cell membranes or introduce antioxidant genes to create an environment that minimizes the decomposition of the final Oxide product.
• Titer and Purity Assays: Validate the engineered strain in fed-batch culture, measuring the final Oxide titer, yield, and stereoisomer purity (HPLC).
• Result Report Output: Compile a detailed Experimental Report including gene modification data, enzyme directed evolution results, and fermentation metrics (volumetric titer and specific Oxide isomer ratio) , 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-efficiency Zeaxanthin Oxide supply. CD Biosynsis provides customized strain and enzyme engineering solutions:

• Detailed Oxidase Kinetic and Selectivity Report , demonstrating the functional enhancement of the Directed Evolution Enzyme Variant.
• Consultation on optimized fermentation conditions (e.g., light exclusion, harvest time) to maximize Oxide stability.
• Experimental reports include complete raw data on total Zeaxanthin Oxide production (mg/L) and final product stability (shelf life testing), essential for cosmetic and pharmaceutical quality control.