Enhancing KAS Expression for Long-Chain Fatty Acid Production
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Solution Overview
Problem
Current methods are inadequate for effectively producing lipids rich in desired long-chain fatty acids, particularly polyunsaturated fatty acids (PUFA), which are essential for nutritional and industrial applications, as they rely on enhancing desaturase or elongase enzymes without specific control over chain length and unsaturation.
Innovation Solution
A method involving the cultivation of a transformant with enhanced expression of a β-ketoacyl-ACP synthase (KAS) gene from Nannochloropsis algae, specifically the NoKASII gene, to improve the productivity of long-chain fatty acids and lipids, including PUFA, by optimizing the fatty acid synthetic pathway.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If desaturase or elongase enzymes are enhanced to improve PUFA productivity, then the production of polyunsaturated fatty acids increases, but the control over chain length and unsaturation levels becomes insufficient
Solution Approach 1:
The patent changes the key parameter from enhancing desaturase/elongase enzymes to enhancing β-ketoacyl-ACP synthase (KAS) enzymes, which control the chain length at the early stage of fatty acid synthesis. This parameter change enables precise control over the carbon chain length (C16, C18, C20) while maintaining high PUFA productivity through coordinated expression with desaturase genes
Solution Approach 2:
The patent applies preliminary action by controlling the chain length determination step (catalyzed by KAS) before the desaturation step. By pre-establishing the correct carbon chain length through KAS enhancement, subsequent desaturase enzymes can then efficiently introduce unsaturated bonds at specific positions, ensuring both productivity and precision
2Adaptability or versatility
If general fatty acid synthesis pathways are used, then various fatty acids are produced, but the production of lipids with specific carbon chain lengths and unsaturation levels is insufficient
Solution Approach 1:
The patent applies local quality by specifically targeting and enhancing the KAS enzyme function within the general fatty acid synthesis pathway. Instead of modifying the entire pathway, the invention focuses on the specific step (chain length determination by KAS) that controls the critical property (carbon chain length), while coordinating with desaturase enhancement for unsaturation control, thereby achieving precise fatty acid composition
3Productivity
If existing lipid production methods are applied, then lipids are produced, but the productivity of lipids rich in desired long-chain fatty acids is insufficient
Solution Approach 1:
The patent changes the approach parameter from enhancing terminal enzymes (desaturase/elongase) to enhancing the early-stage KAS enzyme. This enables more efficient routing of carbon flow toward long-chain fatty acid synthesis by controlling chain elongation at the source, thereby increasing both the productivity and quantity of desired long-chain fatty acids (C18:1, C18:2, C18:3, C20:4, C20:5)
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
Significantly increases the production of long-chain fatty acids and lipids with specific carbon chain lengths and unsaturation levels, such as C18:1, C18:2, C18:3, C20:4, and C20:5, enhancing the fatty acid composition and productivity in microorganisms like Nannochloropsis oculata.
Implementation Method 1
A β-ketoacyl-ACP synthase (hereinafter, also referred to as "KAS") is an enzyme involved in control of chain length of the acyl group
Implementation Method 2
culturing a transformant wherein the expression of a gene encoding the following protein (A) or (B) is enhanced, and producing fatty acids or lipids containing these fatty acids as components
Data Source
AI summary
A method of producing lipids, containing the steps of:culturing a transformant in which the expression of a gene encoding the following protein (A) or (B) is enhanced, andproducing long-chain fatty acids or the lipids containing these fatty acids as components, wherein:protein (A) is a protein consisting of the amino acid sequence set forth in SEQ ID NO: 1; andprotein (B) is a protein consisting of an amino acid sequence having 70% or more identity with the amino acid sequence of the protein (A), and having β-ketoacyl-ACP synthase activity.