Host-Cell Carotenoid Biosynthesis for Pure Apocarotenoid Production
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Solution Overview
Problem
The supply of natural ionones and retinoids is severely limited by their extremely low abundance in nature, and chemically synthesized apocarotenoids often consist of mixed isomers, which are less desirable due to their different odors and higher costs.
Innovation Solution
A method for producing carotenoids and apocarotenoids by expressing optimized enzymes in a host cell using expression vectors linked to promoters, employing optimized gene products and enzymes such as LcyE and CCD1 to enhance production yields and purity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If extraction from natural sources is used, then natural ingredients are obtained, but supply is severely limited and costs are high
Solution Approach 1:
The patent uses genetically engineered microorganisms to produce apocarotenoids through biosynthesis, creating a biological copy of the natural production system. This allows mass production of single isomer compounds (e.g., α-ionone, β-ionone, retinol) without relying on rare natural sources, thereby resolving the contradiction between limited natural supply and the need for abundant natural-like ingredients
Solution Approach 2:
The patent modifies metabolic pathways in host cells by introducing and optimizing specific enzyme genes (e.g., CCD1, LcyE, crtY, blh) to control the biosynthesis of apocarotenoids. By changing the genetic and metabolic parameters of the host organism, the system achieves high-yield production of single isomer compounds that mirror natural products but can be manufactured at scale
2Productivity
If chemical synthesis is used, then production yield is improved, but mixed isomers are produced with different odors
Solution Approach 1:
The patent replaces chemical synthesis methods with biological synthesis using engineered microorganisms. The biochemical pathways mediated by specific enzymes (e.g., carotenoid cleavage oxygenases like CCD1) naturally produce single isomer apocarotenoids with high stereospecificity. This biological approach maintains manufacturing precision for isomer purity while achieving high productivity through microbial fermentation
Solution Approach 2:
The patent employs specific enzyme intermediaries (e.g., CCD1 from Petunia hybrida, LcyE from Lactuca sativa) that act as biological catalysts to convert carotenoid precursors into single isomer apocarotenoids. These enzyme intermediaries provide the necessary stereochemical control to produce pure isomers, avoiding the mixed isomer problem of chemical synthesis
3Productivity
If conventional microbial production is used, then production is achieved, but yields are very low
Solution Approach 1:
The patent segments the metabolic pathway into distinct functional modules: (1) carotenoid biosynthesis pathway (genes like crtY, LcyE), (2) carotenoid cleavage pathway (genes like CCD1, blh), and (3) product extraction system. This segmentation allows independent optimization of each pathway component, enabling high yield production while managing the complexity through modular design
Solution Approach 2:
The patent performs preliminary genetic engineering to create host cells with optimized metabolic pathways before actual production. By pre-introducing and optimizing the necessary enzyme genes (e.g., overexpressing CCD1 and LcyE, modifying promoter regions), the system is prepared to achieve high yields during fermentation, avoiding the need for complex real-time adjustments
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This method achieves high yields of single isomer apocarotenoids like α-ionone and β-ionone, and retinol, overcoming natural supply limitations and consumer preferences for natural ingredients.
Implementation Method 1
expressing in a host cell an expression module comprising an expression vector having a coding region encoding at least one optimised carotenoid or apocarotenoid generating enzyme
Data Source
AI summary
A method for producing a carotenoid or apocarotenoid is disclosed. The method comprises the step of expressing in a host cell an expression module comprising an expression vector having a coding region encoding at least one optimised carotenoid or apocarotenoid generating enzyme, the coding region being operably linked to a promoter. A host cell comprising an expression vector having a coding region encoding at least one optimised carotenoid or apocarotenoid generating enzyme, the coding region being operably linked to a promoter is also provided together with a kit.