Recombinant Camelina EPA DHA Production via FAE1 Deletion
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Solution Overview
Problem
Current methods for producing recombinant plants with high levels of omega-3 long chain polyunsaturated fatty acids (LCPUFAs) like EPA and DHA are limited by high levels of undesired biosynthetic intermediates such as gamma-linolenic acid (GLA) and low production yields, and there is a need for a land-based source of astaxanthin.
Innovation Solution
A recombinant plant expressing a nucleic acid construct comprising Δ6-elongase, Δ5-desaturase, Δ6-desaturase, and optionally other desaturases, with reduced expression of very long chain fatty acid (VLCFA) genes, such as FAE1, to increase EPA and DHA production while decreasing GLA levels, and simultaneously producing astaxanthin in the same oilseed.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If recombinant plants are engineered to produce high levels of omega-3 LCPUFAs (EPA and DHA), then the nutritional value and commercial importance are improved, but high levels of undesired biosynthetic intermediates (GLA) are generated and production yields remain limited
Solution Approach 1:
The patent removes the harmful FAE1 gene (fatty acid elongase 1) from the plant genome through gene editing. This gene is responsible for producing unwanted very long chain fatty acids and GLA intermediates. By extracting/deleting this specific gene, the patent eliminates the source of harmful byproducts while preserving the beneficial omega-3 LCPUFA production pathway.
Solution Approach 2:
The patent modifies the enzymatic parameters of the fatty acid biosynthesis pathway by introducing specific enzyme variants (such as D6D from Phaeodactylum tricornutum, D6E from Physcomitrella patens, and D5D from Phaeodactylum tricornutum) that have altered substrate specificities and reaction kinetics. These parameter changes enable the pathway to favor production of EPA and DHA while reducing formation of GLA and other unwanted intermediates.
2Quantity of substance
If recombinant plants are engineered to produce high levels of omega-3 LCPUFAs, then the nutritional value is improved, but the production yield and efficiency are limited
Solution Approach 1:
The patent ensures continuous flux through the beneficial pathway by introducing multiple enzyme activities (Δ6-elongase, Δ5-desaturase, Δ6-desaturase, w3-desaturase) that work in sequence to convert ALA → EPA → DHA continuously. The pathway is designed to operate at high capacity throughout seed development, maximizing the conversion of substrate fatty acids into valuable omega-3 LCPUFAs.
Solution Approach 2:
The patent optimizes pathway parameters by introducing enzymes with enhanced catalytic activities and substrate affinities. Specifically, the enzyme constructs used (such as D6D from P. tricornutum and D5D from P. tricornutum) have been selected for their high efficiency in converting substrates, thereby increasing the overall productivity of the pathway.
3Quantity of substance
If recombinant plants are engineered to produce omega-3 LCPUFAs, then the nutritional value is improved, but the complexity of the genetic construct and metabolic pathway increases
Solution Approach 1:
The patent divides the complex fatty acid modification pathway into separate functional modules, each encoded by a specific gene: D6D (Δ6-desaturase), D6E (Δ6-elongase), D5D (Δ5-desaturase), and w3D (w3-desaturase). This segmentation allows for independent optimization and expression control of each enzyme activity, making the overall system more manageable and easier to engineer.
Solution Approach 2:
The patent uses a multi-functional enzyme construct that combines several activities (Δ6-elongase, Δ5-desaturase, Δ6-desaturase, and w3-desaturase) in a single genetic framework. This multi-functionality reduces the need for multiple separate gene expressions and simplifies the overall genetic construct while maintaining high productivity.
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
The solution achieves significantly higher levels of EPA and DHA, reduced GLA levels, and increased astaxanthin production, resulting in a more efficient and sustainable production of omega-3 fatty acids and astaxanthin in recombinant plants, enhancing their nutritional and commercial value.
Implementation Method 1
genetic modification (GM) is used to introduce the non-native biosynthetic pathway for omega-3 LC-PUFAs into the nuclear genome of a suitable oilseed host, enabling the plant to convert endogenous C18 fatty acids into the more desirable C20+LC-PUFAs such as eicosapentaenoic acid (EPA; 20:5Δ5,8,11,14,17) and docosahexaenoic acid (DHA; 22:6Δ4,7,10,13,16,19)
Implementation Method 2
The invention relates to improved recombinant plants or plant seeds with increased levels of omega-3 long chain polyunsaturated fatty acids (LCPUFAs) such as eicospentaenoic acid (EPA) and docosahexaenoic acid (DHA)
Data Source
AI summary
The present invention relates to improved recombinant plants or plant cells for producing omega-3 long chain polyunsaturated fatty acids such as eicospentaenoic acid (EPA) and docosahexaenoic acid (DHA). The invention further relates to the oil produced by said recombinant oilseed plant or cell.