Yeast Oleic Acid Yield via Enzyme Segmentation
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Solution Overview
Problem
The existing methods for modulating the lipid profile of oleaginous yeast, such as Yarrowia lipolytica, to increase specific fatty acid content, like oleic acid, are unpredictable and often ineffective, as seen with overexpressing genes like DGA1 and DGA2 without significant impact on lipid content.
Innovation Solution
Genetic modifications are introduced in yeast cells to increase or decrease the activity of specific proteins involved in lipid synthesis and degradation pathways, such as Δ9 desaturase, elongase, and diacylglycerol acyltransferases, through nucleic acid transformations that encode these proteins or their variants, allowing for targeted modulation of lipid composition.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If genes like DGA1 and DGA2 are overexpressed to increase lipid content, then the intent is to improve lipid yield, but the actual effect is unpredictable and often insignificant
Solution Approach 1:
The patent segments the lipid synthesis pathway into multiple enzymatic steps and targets specific enzymes (Δ9 desaturase, elongase, DGAT) for individual modulation rather than attempting to overexpress multiple genes simultaneously. This segmented approach allows precise control over specific biochemical reactions to achieve predictable lipid profile changes.
Solution Approach 2:
The patent applies local quality by selectively modulating specific enzymes at specific positions in the lipid synthesis pathway. Instead of uniform overexpression of all lipid-related genes, the invention targets particular enzymes (e.g., Δ9 desaturase for oleic acid production) to achieve localized functional enhancement with predictable outcomes.
2Quantity of substance
If multiple genes are overexpressed to modulate lipid profile, then the goal is to increase specific fatty acid content, but the outcome is unpredictable and often ineffective
Solution Approach 1:
The patent extracts and isolates specific enzymatic functions from the complex lipid synthesis pathway. By focusing on individual enzymes like Δ9 desaturase, elongase, and DGAT, the invention extracts the critical control points that determine fatty acid composition, enabling precise manipulation of lipid profiles without the unpredictability of multi-gene overexpression.
Solution Approach 2:
The patent employs parameter changes by modifying the activity levels of specific enzymes through targeted genetic modifications. By adjusting the expression level or activity of individual enzymes (e.g., increasing Δ9 desaturase activity to enhance oleic acid production), the invention achieves precise control over lipid composition parameters with predictable results.
3Productivity
If genetic modifications are made to increase enzyme activity, then the intent is to enhance lipid production efficiency, but the results are inconsistent
Solution Approach 1:
The patent applies preliminary action by identifying and preparing specific enzyme candidates (Δ9 desaturase, elongase, DGAT) before implementing genetic modifications. This pre-selection of targeted enzymes based on their known roles in lipid synthesis ensures that subsequent genetic modifications produce consistent and reliable effects on lipid production efficiency.
Data Source
AI summary
Disclosed are transformed cells comprising one or more genetic modifications that affect the lipid content of the cell, e.g., by increasing the concentration of oleic acid in the cell relative to an unmodified cell of the same type. Also disclosed are methods for modifying the lipid content of a cell by increasing the activity of one or more proteins in the cell and/or by decreasing the activity of one or more proteins in the same cell.