Carotenoid Esterase Genes for Stable Plant Carotenoid Esterification
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Solution Overview
Problem
The genes encoding carotenoid esterase in Lycium barbarum, which are crucial for catalyzing the esterification of free carotenoids, have not been reported, leading to instability and reduced efficacy of carotenoids due to oxidation and environmental conditions.
Innovation Solution
Introduction of LbZAT1, LbZAT2, and LbZAT3 genes, which encode carotenoid esterase proteins, into plant host cells to catalyze the esterification of free carotenoids, enhancing their stability and lipid solubility.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If free carotenoids are used in plant tissues, then they can be easily obtained and synthesized, but they are prone to oxidation and E/Z tautomerism under high temperature or UV conditions, reducing their stability
Solution Approach 1:
The patent changes the chemical parameter of carotenoids by introducing esterification reactions. The carotenoid esterase enzyme catalyzes the conversion of free carotenoids to esterified carotenoids, fundamentally altering the chemical structure to improve stability. This parameter change (from free hydroxyl to esterified form) directly addresses the oxidation and tautomerism problems.
Solution Approach 2:
The carotenoid esterase enzyme acts as an intermediary that facilitates the esterification of carotenoids. By introducing this enzymatic mediator, the patent enables controlled conversion of unstable free carotenoids into stable esterified forms, resolving the stability issue without direct exposure to harsh conditions that cause oxidation.
2Reliability
If carotenoids are esterified to enhance lipid solubility and stability, then their tolerance to adverse environmental conditions improves, but the complexity of the esterification process increases
Solution Approach 1:
The patent employs the carotenoid esterase enzyme to enable self-service esterification within the plant system. The enzyme is introduced into plant cells where it autonomously catalyzes the esterification of carotenoids using endogenous fatty acid substrates, eliminating the need for complex external esterification equipment or multi-step processing procedures.
Solution Approach 2:
The carotenoid esterase serves as a biological intermediary that simplifies the esterification process. Rather than requiring complex chemical esterification equipment and procedures, the enzyme mediates the reaction in vivo, converting free carotenoids to esterified forms through a straightforward biochemical pathway that occurs naturally within the plant cells.
3Adaptability or versatility
If free carotenoids are distributed in plant tissues, then they can be easily accessed, but they tend to be distributed in the periphery of liposomes or phospholipid layer, limiting their application potential
Solution Approach 1:
The patent changes the physical-chemical parameters of carotenoids through esterification, which alters their solubility characteristics and membrane permeability. Esterified carotenoids with increased lipid solubility can penetrate and localize within the interior of liposomes and microsomes, fundamentally changing their spatial distribution and enabling new applications in drug delivery and nutraceuticals.
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
Significantly increases the content of zeaxanthin dipalmitate and alters carotenoid metabolism, making esterified carotenoids more stable and suitable for applications in liposomes and microsomes.
Implementation Method 1
catalyzes the esterification of free carotenoids
Implementation Method 2
carotenoid esterase... catalyzes the esterification
Data Source
AI summary
The present disclosure discloses three carotenoid esterases and the application of their encoding genes. The genes encoding the carotenoid esterase include: LbZAT1 gene with the nucleotide sequence as shown in SEQ ID NO:1, LbZAT2 gene with the nucleotide sequence as shown in SEQ ID NO:2, and/or LbZAT3 gene with the nucleotide sequence as shown in SEQ ID NO:3. The proteins encoded by the LbZAT1, LbZAT2 and LbZAT3 genes of the present disclosure have the function of catalyzing the esterification reaction between carotenoids containing free hydroxyl and fatty acid acyl donors. Carotenoid esterase and its encoding genes have important application value in the biosynthesis of esterified carotenoids in vitro.