Oxidoreductase Variant Engineering for Higher L-Glutamic Acid Yield
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Solution Overview
Problem
Existing methods for enhancing L-glutamic acid production in microorganisms such as Corynebacterium sp. lack effective strategies to increase productivity through targeted modifications of enzymes involved in the biosynthetic pathway.
Innovation Solution
Introduction of an oxidoreductase variant with a specific amino acid substitution, such as serine to asparagine at position 321, and a polynucleotide encoding this variant, integrated into a transformant to enhance the enzymatic activity and productivity of L-glutamic acid production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If wild-type strains or mutant strains are used for L-glutamic acid production, then production capability is achieved, but productivity is insufficient compared to recombinant strains with engineered enzymes
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of the oxidoreductase enzyme, specifically substituting serine with asparagine at position 321. This enzymatic parameter modification optimizes the carbon source flux through the biosynthetic pathway, thereby increasing L-glutamic acid productivity without requiring complex multi-gene engineering approaches
Solution Approach 2:
The invention focuses on local quality by targeting a specific region of the oxidoreductase enzyme (position 321 in the amino acid sequence) for modification. This localized change in enzyme structure optimizes its catalytic activity and substrate specificity, improving productivity without altering the entire biosynthetic pathway or multiple genes
2Productivity
If multiple genes and proteins involved in biosynthetic pathway are modified to increase L-glutamic acid production, then productivity may improve, but the complexity and difficulty of research and modification increase
Solution Approach 1:
The patent extracts the key limiting factor from the complex biosynthetic pathway by identifying and modifying only the oxidoreductase enzyme. This extraction approach isolates the critical enzymatic step that controls carbon source flux, allowing productivity improvement without attempting to modify all dozens of proteins and genes involved in the pathway
Solution Approach 2:
The invention applies segmentation by dividing the biosynthetic pathway into distinct functional steps and identifying the rate-limiting step catalyzed by oxidoreductase. By focusing modification efforts on this specific segmented portion of the pathway rather than the entire complex system, the patent achieves productivity improvement with minimal modification 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 modified oxidoreductase variant increases L-glutamic acid production by up to 12.1% compared to parent strains, demonstrating improved carbon source flux in the biosynthetic pathway.
Implementation Method 1
an enzyme that catalyzes a redox reaction to supply energy necessary for a living body, and acts to oxidize one compound while reducing another compound
Implementation Method 2
since the TCA cycle lacks the metabolic pathway for the oxidation of a-ketoglutaric acid to succinic acid and isocitrate dehydrogenase and glutamate dehydrogenase are closely involved therein
Data Source
AI summary
The present invention relates to a Corynebacterium sp. mutant microorganism producing L-glutamic acid and a method of producing L-glutamic acid using the same, and more specifically, to a novel oxidoreductase variant involved in the L-glutamic acid biosynthetic pathway, a polynucleotide, and a transformant, as well as a method of producing L-glutamic acid using the same. The oxidoreductase variant according to the present invention is obtained by substituting one or more amino acids in the amino acid sequence constituting oxidoreductase to change the enzymatic activity of the oxidoreductase, and a recombinant microorganism comprising the oxidoreductase variant is capable of efficiently producing L-glutamic acid.