Transgenic Organism Fatty Acid Synthesis Pathway Engineering
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Solution Overview
Problem
Current methods for producing polyunsaturated fatty acids in organisms, such as transgenic plants and microorganisms, face challenges in achieving high yields and specificity, particularly in converting ω6-fatty acids to ω3-fatty acids, due to limitations in desaturase and elongase enzymes, which result in low content and mixed fatty acid production.
Innovation Solution
Introducing nucleic acid sequences encoding Δ5-elongase, Δ5-desaturase, Δ4-desaturase, Δ12-desaturase, and ω3-desaturase activities into organisms to shift the fatty acid synthesis pathway from ω6 to ω3, using enzymes derived from Thalassiosira, Euglena, and Ostreococcus, to produce polyunsaturated fatty acids with enhanced specificity and yield.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional desaturase and elongase enzymes are used in transgenic organisms, then fatty acid production occurs, but the yield and specificity of ω3-fatty acids are low due to mixed fatty acid production
Solution Approach 1:
The patent divides the fatty acid synthesis pathway into multiple discrete enzymatic steps, introducing separate genes for Δ5-elongase, Δ5-desaturase, Δ4-desaturase, Δ12-desaturase, and ω3-desaturase. This segmentation allows each enzyme to be optimized for specific substrate conversion, eliminating mixed fatty acid production and achieving high specificity for ω3-fatty acids while maintaining high yield through coordinated action of specialized enzymes.
2Productivity
If existing desaturase enzymes are introduced into organisms, then some polyunsaturated fatty acid production is achieved, but the conversion of ω6-fatty acids to ω3-fatty acids remains inefficient
Solution Approach 1:
The patent changes the enzymatic parameters by introducing multiple specialized enzymes with different substrate specificities and catalytic properties. The Δ5-elongase and Δ5-desaturase enzymes are specifically optimized for converting C20:4 and C22:4 ω6-fatty acids to their ω3 counterparts, while the Δ12-desaturase handles C18:2 conversion. This parameter optimization achieves efficient and specific conversion of ω6 to ω3 fatty acids, overcoming the limitations of conventional single-enzyme systems.
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 process achieves a significant increase in polyunsaturated fatty acid production, particularly ω3-fatty acids like EPA and DHA, with improved specificity and yield, reaching concentrations of at least 10% by weight in transgenic organisms, and allows for the production of these fatty acids as pure products or in mixtures, suitable for food and pharmaceutical applications.
Implementation Method 1
nucleic acid sequences which encode polypeptides with Δ5-elongase activity... These desaturases and elongases are advantageously derived from Thalassiosira, Euglena or Ostreococcus
Data Source
AI summary
The invention relates to a method for the production of multiply-unsaturated fatty acids in an organism, into which nucleic acids have been introduced, which code for polypeptides with Δ-5 elongase activity. Said nucleic acid sequences, optionally with further nucleic acid sequences, coding for polypeptides for the biosynthesis of fatty acids and lipid metabolism, are advantageously expressed in the organism. Nucleic acid sequences coding for a Δ-6 desaturase, a Δ-5 desaturase, Δ-4 desaturase and/or Δ-6 elongase activity are particularly advantageous and, advantageously, said saturases and elongases are derived from Thalassiosira, Euglena or Ostreococcus. The invention further relates to a method for the production of oils and/or triacylglycerides with an increased content of long-chain, multiply-unsaturated fatty acids. A particular embodiment of the invention is a method for the production of unsaturated ω-3 fatty acids and a method for the production of triglycerides with an increased content of unsaturated fatty acids, in particular, of Δ-3 fatty acids with more than three double bonds. Also disclosed is the production of a transgenic organism, preferably a transgenic plant, or a transgenic microorganism with increased content of ω-3 fatty acids, oils or lipids with ω-3 double bonds as a result of the expression of the elongases and desaturases employed in the above method, preferably in combination with ω-3 desaturases, for example a ω-3 desaturase from fungi of the family Pythiaceae such as the genus Phytophtora, for example, the genus and species Phytophtora infestans, or a ω-3 desaturase from algae such as the family Prasinophyceae, for example, the genus Ostreococcus and, particularly, the genus and species Ostreococcus tauri or diatomaceæ such as the genus Thalassiosira and, particularly, the genus and species Thalassiosira pseudonana. The invention also relates to the nucleic acid sequences, nucleic acid constructs, vectors and organisms containing the nucleic acid sequences and/or the nucleic acid constructs and transgenic organisms containing said nucleic acid sequences, nucleic acid constructs and/or vectors. A further part of the invention relates to oils, lipids and/or fatty acids produced according to the above method and use thereof and, furthermore, unsaturated fatty acids and triglycerides with an increased content of unsaturated fatty acids and use thereof.