Stramenopile Microbial Oil EPA Production via Genetic Pathway Segmentation
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Solution Overview
Problem
Current methods for modifying the fatty acid composition of stramenopile microorganisms, such as those in the class Labyrinthulomycetes, are limited in efficiently producing polyunsaturated fatty acids like eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), as they primarily focus on disrupting polyketide synthase genes without addressing the elongase/desaturase pathway.
Innovation Solution
The method involves disrupting and inhibiting genes associated with fatty acid biosynthesis, specifically targeting fatty acid chain elongase and desaturase genes, and introducing desaturase genes to enhance the production of unsaturated fatty acids in stramenopile microorganisms like Thraustochytrium and Schizochytrium species, thereby modifying their fatty acid composition to accumulate high levels of EPA and other polyunsaturated fatty acids.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If polyketide synthase genes are disrupted to produce polyunsaturated fatty acids, then EPA and DHA production is improved, but the elongase/desaturase pathway is not optimized leading to limited efficiency
Solution Approach 1:
The patent divides the fatty acid biosynthesis pathway into distinct segments: polyketide synthase genes, elongase genes, and desaturase genes. By disrupting polyketide synthase genes and separately introducing exogenous elongase and desaturase genes, the invention segments the complex biosynthetic pathway into manageable modules that can be independently optimized, thereby improving EPA and DHA production efficiency while managing pathway complexity.
Solution Approach 2:
The patent combines multiple genetic modifications into a unified approach: disrupting endogenous polyketide synthase genes while simultaneously introducing exogenous elongase and desaturase genes. This merging of multiple genetic engineering strategies creates a synergistic effect that overcomes the limitations of single-gene disruption and achieves high-level polyunsaturated fatty acid production.
2Quantity of substance
If genes associated with fatty acid biosynthesis are disrupted to modify fatty acid composition, then unsaturated fatty acid content is enhanced, but production efficiency is reduced
Solution Approach 1:
The patent changes the genetic parameters of the fatty acid biosynthesis pathway by disrupting specific polyketide synthase genes and introducing exogenous elongase and desaturase genes with different functional characteristics. This parameter change approach modifies the enzymatic activities and gene expression levels to achieve both high unsaturated fatty acid content and maintained production efficiency through optimized pathway flux.
Solution Approach 2:
The patent applies local quality modification by specifically targeting and disrupting only the polyketide synthase genes responsible for producing less desirable fatty acids, while preserving and enhancing other parts of the biosynthetic pathway through introduced elongase and desaturase genes. This localized genetic modification ensures high unsaturated fatty acid content without compromising overall production efficiency.
Data Source
AI summary
A method for producing a microbial oil includes the steps of: genetically modifying a labyrinthulid by disrupting and/or silencing a gene, or by transforming another gene in addition to the disruption and/or gene silencing of the gene; culturing the labyrinthulid, such that a fatty acid composition accumulated in the labyrinthulid comprises an increased EPA content; and collecting the microbial oil having the increased EPA content from the labyrinthulid. The increased EPA content is not less than 3.3% of a total fatty acid composition.