Mutated OrfB Protein for Microbial EPA Production
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Solution Overview
Problem
Current methods for industrially producing eicosapentaenoic acid (EPA) are inefficient due to high by-product contamination and low production efficiency, particularly when purifying EPA from fish oil.
Innovation Solution
A microorganism is engineered to produce EPA by introducing mutations into specific amino acid residues of the OrfB protein, allowing it to produce EPA at high concentrations, either by expressing mutated OrfB in microorganisms capable of producing docosahexaenoic acid (DHA) or by introducing DHA metabolic pathway genes into organisms without this capability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If EPA is purified from fish oil using conventional methods, then EPA can be obtained, but the production efficiency is low and by-product contamination is high
Solution Approach 1:
The invention changes the biological parameters of the microorganism by mutating the OrfB protein's amino acid residues, transforming it from a DHA-producing organism to an EPA-producing organism. This fundamental parameter change in the biological system enables direct EPA production without the need for purification from fish oil, thereby resolving the contradiction between production efficiency and by-product contamination
Solution Approach 2:
The invention replaces the mechanical/chemical purification process with a biological production system. Instead of extracting and purifying EPA from fish oil through complex separation processes, the patent uses genetically modified microorganisms to directly synthesize EPA, substituting the entire purification mechanism with a targeted biosynthetic pathway
2Productivity
If mutations are introduced into OrfB protein to change product specificity from DHA to EPA, then EPA production efficiency increases, but the complexity of strain development increases
Solution Approach 1:
The invention applies local quality change by mutating only specific amino acid residues (positions 6, 65, 230, 231, and 275) of the OrfB protein rather than redesigning the entire protein structure. This localized modification approach enables product specificity change from DHA to EPA while minimizing the overall complexity of strain development
Solution Approach 2:
The invention changes specific parameters of the OrfB protein (amino acid sequence at defined positions) to alter the enzyme's substrate specificity and product outcome. By focusing parameter changes on critical residues rather than comprehensive protein redesign, the patent achieves high EPA production efficiency with controlled development complexity
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
The method enables efficient production of EPA with reduced by-products, achieving high yields and low production costs, making it suitable for industrial-scale production.
Implementation Method 1
a microorganism having an ability to produce DHA, wherein the microorganism contains a protein composed of an amino acid sequence in which at least one of the amino acid residues at positions 6, 65, 230, 231, and 275 in the amino acid sequence represented by SEQ ID NO: 2 has been substituted with another amino acid residue
Data Source
AI summary
An object of the present invention is to provide a microorganism that efficiently produces EPA and a method for producing EPA using the microorganism. The present invention relates to a microorganism having an ability to produce docosahexaenoic acid (DHA), wherein the microorganism contains a protein composed of an amino acid sequence in which at least one of the amino acid residues at positions 6, 65, 230, 231, and 275 in the amino acid sequence represented by SEQ ID NO: 2 has been substituted with another amino acid residue (mutated OrfB), and is capable of producing eicosapentaenoic acid (EPA), and the like.