Enhancing Factor Proteins for PUFA Synthase Activity
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Solution Overview
Problem
Current methods for producing polyunsaturated fatty acids (PUFAs) in genetically modified organisms do not achieve the same levels of production and DHA:DPAn-6 ratio as seen in native organisms like Schizochytrium 20888, indicating a need for additional factors to enhance PUFA synthase activity.
Innovation Solution
Identification and expression of enhancing factor proteins, such as Sz-TE2 and Sz-TE3, which are at least 90% identical to specific amino acid sequences, to enhance the enzymatic activity of PUFA synthases in host organisms, thereby increasing PUFA production and altering the DHA:DPAn-6 ratio.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If PUFA synthase systems are introduced into heterologous host organisms, then PUFA production capability is achieved, but production levels and DHA:DPAn-6 ratios remain insufficient compared to native organisms
Solution Approach 1:
The patent introduces enhancing factor proteins as intermediary components that mediate between the PUFA synthase system and the host organism's cellular environment. These factor proteins (encoded by genes such as sz-TE2 and sz-TE3) act as mediators to optimize the function of PUFA synthase in heterologous hosts, thereby improving both production levels and fatty acid composition ratios to match native organism performance
2Adaptability or versatility
If PUFA synthase systems are expressed in heterologous hosts, then PUFA synthesis capability is obtained, but the DHA:DPAn-6 ratio is altered and does not match native organisms
Solution Approach 1:
The patent employs parameter changes by introducing enhancing factor proteins that modify the operational parameters of PUFA synthase in heterologous hosts. These factor proteins adjust enzymatic activity parameters and substrate utilization parameters to restore the native DHA:DPAn-6 ratio, allowing the system to maintain fidelity to native organism composition while functioning in diverse host organisms
3Ease of manufacture
If only PUFA synthase genes are introduced into host organisms, then basic PUFA production is achieved, but production levels fall short of commercial requirements
Solution Approach 1:
The patent merges multiple functional components into a integrated genetic construct: PUFA synthase genes are combined with enhancing factor protein genes (such as sz-TE2 and sz-TE3) in a single genetic modification strategy. This merged approach maintains relative ease of manufacture through single-step or co-expression system transformation while achieving synergistic effects that dramatically improve PUFA accumulation levels beyond what either component could achieve alone
Data Source
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AI summary
Factors For the Production and Accumulation of Polyunsaturated Fatty Acids (PUFAs) Derived from PUFA Synthases Abstract Disclosed are novel enhancing factor proteins of the PUFA synthase systems, nucleic acid molecules encoding the same, recombinant nucleic acid molecules and recombinant host cells comprising such nucleic acid molecules, genetically modified microorganisms comprising the same, and methods of making and using the same. Also disclosed are genetically modified microorganisms that have been genetically modified to express a PUFA synthase system for the production of PUFAs, wherein the microorganisms have been modified to express the novel enhancing factor proteins of the PUFA synthase system.