Microalgae PFA1 PFA3 Overexpression EPA Production
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Solution Overview
Problem
Current methods for producing omega-3 polyunsaturated fatty acids, such as eicosapentaenoic acid (EPA), in microalgae are limited by low production levels and undesirable fatty acid profiles, making it challenging to achieve significant and consistent production of EPA and other essential fatty acids like DHA.
Innovation Solution
Modifying microalgae to alter the expression levels of genes encoding polyunsaturated fatty acid synthase subunits, specifically PFA1 and PFA3, to increase EPA production and manipulate the EPA:DHA ratio, by overexpressing these genes and using promoter engineering to enhance expression levels.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional methods are used for producing omega-3 PUFAs in microalgae, then production can be maintained at baseline levels, but production levels remain low and fatty acid profiles are undesirable
Solution Approach 1:
The patent changes the expression levels of PUFA synthase subunit genes (PFA1, PFA2, PFA3) as key parameters to control fatty acid synthesis. By overexpressing specific subunits, the patent achieves up to 20-fold increase in EPA production while simultaneously improving the fatty acid profile to contain higher omega-3 PUFAs and lower omega-6 PUFAs, thus resolving the contradiction between productivity and profile quality
Solution Approach 2:
The patent segments the PUFA synthase system into three separate subunits (PFA1, PFA2, PFA3) that can be independently manipulated. This segmentation allows selective overexpression of specific subunits to optimize both EPA production levels and fatty acid profile composition, enabling independent control of productivity and quality parameters
2Productivity
If gene expression levels of PUFA synthase subunits are increased, then EPA production increases significantly, but process complexity increases due to genetic modification requirements
Solution Approach 1:
The patent utilizes the native PUFA synthase system of microalgae, which already performs essential fatty acid synthesis. By introducing expression control mechanisms for the existing PFA1, PFA2, and PFA3 subunits, the system maintains its natural function while achieving enhanced EPA production. This multi-functional approach avoids the need for completely synthetic pathways, reducing overall process complexity
3Manufacturing precision
If PFA1 and PFA3 expression is increased to enhance EPA production, then EPA:DHA ratio improves, but manipulation of multiple genes increases process complexity
Solution Approach 1:
The patent applies local quality control by specifically targeting PFA1 and PFA3 subunits for overexpression to control EPA production, while recognizing that all three subunits (PFA1, PFA2, PFA3) work together in the PUFA synthase complex. This localized approach to gene manipulation allows precise control of EPA:DHA ratio without requiring comprehensive modification of the entire fatty acid synthesis pathway
Data Source
AI summary
The subject disclosure features, in one aspect, a method for producing lipids enriched for EPA, comprising modifying a microalga to increase expression of PFA1, and/or PFA3, and culturing the modified microalga under conditions which allow the expression of PFA1, and/or PFA3, wherein lipids enriched for EPA are produced. Also featured is a recombinant microalga in which PFA1, and/or PFA3, is overexpressed. Such recombinant microalgae have been demonstrated herein to produce very favorable fatty acid lipid profiles (e.g., increased levels of EPA, increased ratio of EPA:DHA, decreased levels of DPA n-6, etc.).