FatB Thioesterase Clades for Specific Fatty Acid Enrichment
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Solution Overview
Problem
Current methods for altering fatty acid profiles in oils produced by microalgae and plants are limited in specificity and efficiency, particularly in achieving targeted enrichment of certain fatty acid chains like C8, C10, C12, C14, and C16.
Innovation Solution
The use of nucleic acid constructs containing FatB genes with specific amino acid sequences, which encode active acyl-ACP thioesterases, to produce altered fatty acid profiles in oils by expressing these genes in host cells such as microalgae and plants, thereby enriching the oils with desired fatty acid chains.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If current methods for altering fatty acid profiles are used, then some fatty acid modification is achieved, but the specificity and efficiency are limited
Solution Approach 1:
The patent applies parameter changes by identifying and utilizing specific amino acid sequence parameters of FatB thioesterase enzymes. By changing the amino acid sequence parameters to match specific clades (clade 1-12), the enzyme achieves higher specificity for particular fatty acid chain lengths while maintaining high catalytic efficiency. This is evidenced by the sequence identity thresholds (e.g., at least 83.8% identity for clade 4, at least 94.6% for clade 5) that define functional specificity.
Solution Approach 2:
The patent applies local quality by creating localized specificity within the enzyme's substrate binding region. Different clades of FatB enzymes have evolved specific amino acid compositions in their active sites that confer preference for particular acyl chain lengths (C8-C16). This localized structural differentiation enables the enzyme to selectively process specific substrates while maintaining overall catalytic efficiency, resolving the contradiction between specificity and productivity.
2Manufacturing precision
If FatB genes with specific amino acid sequences are used, then precise modification of fatty acid profiles is achieved, but the complexity of gene selection and characterization increases
Solution Approach 1:
The patent applies segmentation by dividing the FatB enzyme family into 12 distinct clades based on amino acid sequence homology and functional specificity. Each clade represents a segmented category with defined sequence identity thresholds (e.g., clade 1 requires at least 85.9% identity, clade 2 requires at least 89.5%). This segmentation simplifies the complex task of gene selection by providing a structured classification system that maps specific clades to specific fatty acid chain length preferences, reducing the complexity of predicting and achieving desired fatty acid profiles.
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
This approach allows for precise modification of fatty acid profiles in oils, increasing the mass of specific fatty acids by at least 10% compared to non-enriched oils, making it suitable for various applications in food, chemicals, and fuels.
Implementation Method 1
Type II fatty acid biosynthesis typically involves extension of a growing acyl-ACP (acyl-carrier protein) chain by two carbon units followed by cleavage by an acyl-ACP thioesterase
Data Source
AI summary
The invention features plant acyl-ACP thioesterase genes of the FatB class and proteins encoded by these genes. The genes are useful for constructing recombinant host cells having altered fatty acid profiles. Oleaginous microalga host cells with the new genes or previously identified FatB genes are disclosed. The microalgae cells produce triglycerides with useful fatty acid profiles.