Pex Gene Disruption for PUFA Yield in Oleaginous Yeast
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Solution Overview
Problem
Current methods for producing large-scale quantities of polyunsaturated fatty acids (PUFAs) in organisms like Saccharomyces cerevisiae and other microbial systems face challenges in improving yield and controlling oil composition, as they rely on natural microbial abilities and lack effective optimization for commercial production.
Innovation Solution
Disrupting peroxisome biogenesis factor proteins in oleaginous eukaryotic organisms, such as Yarrowia lipolytica, to increase the weight percent of PUFAs by introducing genes encoding a functional PUFA biosynthetic pathway and disrupting native genes encoding peroxisome biogenesis factor proteins, thereby enhancing PUFA production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If peroxisome biogenesis factor proteins are disrupted in oleaginous eukaryotic organisms, then the weight percent of polyunsaturated fatty acids increases, but the complexity of genetic manipulation increases
Solution Approach 1:
The patent extracts and removes peroxisome biogenesis factor proteins (Pex proteins) from the oleaginous eukaryotic organism through genetic disruption. By taking out these specific proteins that are responsible for peroxisome assembly and function, the patent redirects metabolic pathways to increase PUFA accumulation in the cytosol, thereby resolving the contradiction between increasing PUFA quantity and managing genetic manipulation complexity.
Solution Approach 2:
The patent changes the biological parameter of peroxisome biogenesis by disrupting Pex gene expression. This parameter change in protein synthesis leads to altered metabolic flux, redirecting fatty acid synthesis from peroxisomal beta-oxidation pathways to cytosolic PUFA accumulation, thus achieving increased PUFA weight percent through a fundamental biological parameter change.
2Productivity
If peroxisome biogenesis factor proteins are disrupted to increase PUFA production, then productivity improves, but manufacturing precision becomes more difficult to control
Solution Approach 1:
The patent implements a feedback mechanism where the disruption of Pex proteins creates a metabolic state that naturally favors PUFA accumulation. The altered metabolic flux serves as feedback that redirects carbon flow toward PUFA synthesis pathways, allowing the system to self-regulate and maintain high PUFA productivity while achieving consistent oil composition through the established metabolic rerouting.
3Adaptability or versatility
If genes encoding PUFA biosynthetic pathway are introduced, then the functional capability for PUFA synthesis improves, but the device complexity increases
Solution Approach 1:
The patent merges the introduction of heterologous PUFA biosynthetic genes with the disruption of native Pex proteins in a coordinated genetic modification strategy. By combining these two genetic interventions, the patent creates a synergistic effect where the introduced PUFA pathway genes benefit from the metabolic rerouting caused by Pex disruption, achieving enhanced PUFA synthesis capability while managing overall genetic complexity through integrated design.
Data Source
AI summary
Methods of increasing the amount of polyunsaturated fatty acids (PUFAs) in the total lipid fraction and in the oil fraction of PUFA-producing, oleaginous eukaryotes, accomplished by modifying the activity of peroxisome biogenesis factor (Pex) proteins. Disruptions of a chromosomal Pex3 gene, Pex10p gene or Pex16p gene in a PUFA-producing, oleaginous eukaryotic strain resulted in an increased amount of PUFAs, as a percent of total fatty acids and as a percent of dry cell weight, in the total lipid fraction and in the oil fraction of the strain, as compared to the parental strain whose native Pex protein was not disrupted.