Lycopene Oxide Bioproduction Engineering Service

Lycopene Oxide (Lycopene Epoxide) is a derivative of the powerful antioxidant Lycopene, exhibiting enhanced biological activities and unique applications in pharmaceuticals and cosmetology. Traditional production via chemical synthesis is highly inefficient, characterized by many steps in chemical synthesis , leading to low overall yield and complex purification of multiple isomers. Furthermore, the product itself, Lycopene Oxide, is highly susceptible to degradation, posing the challenge of poor stability during both synthesis and storage. Microbial biosynthesis offers a sustainable, single-step enzymatic approach to overcome these limitations.

CD Biosynsis offers a synthetic biology service focused on efficient Lycopene Oxide production using Saccharomyces cerevisiae (baker's yeast), a robust and GRAS-status host. Our core strategy involves modification of lycopene synthesis pathway in Saccharomyces cerevisiae . We first engineer the yeast's native MVA (Mevalonate) pathway and introduce the complete Lycopene synthesis gene cluster (CrtE, CrtB, CrtI) to achieve high-titer production of the precursor, Lycopene. This foundation is then coupled with directed evolution of oxidase . We identify and introduce a specific Monooxygenase (e.g., a modified CYP enzyme) capable of selectively epoxidizing Lycopene to form Lycopene Oxide. This enzyme is subjected to directed evolution or site-saturation mutagenesis to enhance its activity (kcat), substrate affinity (Km), and, critically, its regioselectivity for the desired epoxide isomer. This integrated approach aims to deliver a high-yield, specific Lycopene Oxide product from a food-grade microbial host, simplifying downstream processing and ensuring high product quality and stability.

Pain Points

Efficient and stable Lycopene Oxide production faces these key challenges:

• Many Steps in Chemical Synthesis: Chemical epoxidation involves multiple intermediate reactions, toxic catalysts, and harsh conditions , resulting in low overall yield and high production costs.
• Poor Stability: Lycopene Oxide, like its precursor Lycopene, is highly sensitive to light, heat, and oxidation , leading to rapid degradation and loss of biological activity during manufacture and storage.
• Low Lycopene Precursor Supply: The native MVA pathway in S. cerevisiae is tightly regulated. Simply introducing the Crt genes often leads to a bottleneck in precursor supply , limiting Lycopene yield.
• Low Epoxidation Specificity: Non-enzymatic or non-optimized chemical oxidation produces a complex mixture of mono-, di-, and tri-epoxides and various isomers , making purification of the desired isomer nearly impossible.

A successful solution must ensure high-yield Lycopene precursor production and achieve highly selective, enzymatic conversion to the target epoxide.

Solutions

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

Modification of Lycopene Synthesis Pathway in S. cerevisiae

           

We upregulate the native MVA pathway (e.g., tHMG1 gene) and optimize the Crt gene cluster expression to maximize flux towards the Lycopene precursor.

Directed Evolution of Oxidase

We use error-prone PCR or site-saturation mutagenesis to screen CYP450 variants for enhanced catalytic activity and high regioselectivity for Lycopene epoxidation.

CYP450 Reductase and Partner Optimization

We co-express the modified CYP450 with an optimized reductase partner (e.g., CPR) to ensure sufficient electron supply, maximizing the oxidase reaction rate.

Metabolic Sink Deletion

We knock out competing pathways (e.g., the synthesis of Ergosterol) that drain carbon flux from the MVA pathway, thereby increasing the effective Lycopene yield.

This systematic approach ensures efficient precursor synthesis and highly selective enzymatic conversion to the final product.

Advantages

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

High Isomer Purity

Enzymatic epoxidation achieves a specific mono-epoxide isomer with minimal unwanted by-products, solving the chemical synthesis issue.

Reduced Degradation

Production in the yeast host allows for in situ encapsulation (e.g., in lipid bodies), protecting the unstable product from environmental stress.

Green and Cost-Effective Route

Eliminating multiple chemical steps, toxic reagents, and high-energy inputs lowers both pollution and manufacturing costs .

High Titer from Enhanced MVA Pathway

Metabolic flux rerouting ensures a massive supply of the Isoprenoid precursor , leading to high final product yield.

Food-Grade GRAS Host Icon

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

We provide a sustainable and competitive biosynthetic route for high-quality Lycopene Oxide production.

Process

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

• Lycopene Pathway Assembly: Overexpress the yeast MVA pathway (tHMG1) and integrate the complete Lycopene cluster (CrtE, CrtB, CrtI) to establish the precursor strain.
• Oxidase Discovery and Mutagenesis: Identify initial candidate Monooxygenases and perform directed evolution or rational design to create variants with improved Lycopene specificity.
• Co-expression Optimization: Co-express the evolved oxidase with the Lycopene pathway and optimize the expression levels of the CYP450 reductase partner (CPR).
• Flux Rerouting: Knock out or downregulate competing pathways (e.g., Ergosterol synthesis, SQS) to maximize carbon availability for Lycopene production.
• Fermentation Performance Validation: Test the final engineered strain in fed-batch fermentation to assess Lycopene Oxide titer, yield, and epoxide purity (ratio of mono-epoxide to di-epoxide).
• 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 high-purity, stable Lycopene Oxide supply. CD Biosynsis provides customized strain and enzyme engineering solutions:

• Detailed Epoxide Specificity and Titer Report , demonstrating the single-step conversion efficiency.
• Consultation on optimized cell disruption and extraction protocols tailored for Lycopene Oxide recovery from yeast biomass.
• Experimental reports include complete raw data on total Lycopene Oxide production (mg/L) and the mono-epoxide fraction (%) , essential for pharmaceutical quality control.