Fermentation Purity via tyrB Gene Attenuation
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Solution Overview
Problem
Current methods for producing 2-methyl-butyric acid by fermentation of bacteria belonging to the order Enterobacterales result in high levels of byproduct substances, which complicates the production of highly pure 2-methyl-butyric acid at a low cost.
Innovation Solution
Attenuating the expression of the tyrB gene in a 2-methyl-butyric acid-producing bacterium belonging to the order Enterobacterales reduces the production of byproduct substances such as 3-methyl-butyric acid, isobutyric acid, L-allo-isoleucine, and D-allo-isoleucine, thereby enhancing the purity and reducing the production costs of 2-methyl-butyric acid.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional fermentation methods are used to produce 2-methyl-butyric acid, then production cost is reduced, but the purity of 2-methyl-butyric acid deteriorates due to high levels of byproduct substances
Solution Approach 1:
The patent extracts and eliminates the harmful byproduct formation pathway by deleting the tyrB gene, which encodes tyrosine aminotransferase. This specific gene deletion removes the metabolic route that produces unwanted byproducts like 3-methyl-butyric acid, isobutyric acid, and allo-isoleucine, thereby extracting the harmful element from the fermentation system while maintaining 2-methyl-butyric acid production.
Solution Approach 2:
The patent changes the metabolic parameter of the bacterium by modifying its genetic composition - specifically deleting the tyrB gene. This genetic parameter change fundamentally alters the metabolic profile of the organism, shifting it from producing high levels of byproducts to producing high purity 2-methyl-butyric acid, thus resolving the purity-cost contradiction.
2Manufacturing precision
If the tyrB gene is attenuated to reduce byproduct substances, then the purity of 2-methyl-butyric acid is improved, but the complexity of the fermentation process increases
Solution Approach 1:
The complexity issue is resolved by extracting only the specific tyrB gene responsible for byproduct formation, rather than implementing complex multi-step purification processes or multiple gene modifications. This single gene deletion approach simplifies the overall process complexity while achieving the desired purity improvement.
3Quantity of substance
If conventional fermentation is used, then the production process is simple, but the amount of byproduct substances increases
Solution Approach 1:
The patent successfully extracts and removes the harmful byproduct formation pathway through tyrB gene deletion, achieving a significant reduction in byproduct substances (3-methyl-butyric acid, isobutyric acid, L-allo-isoleucine, and D-allo-isoleucine) while maintaining the simplicity of the fermentation process with only a single genetic modification required.
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 bacterium produces 2-methyl-butyric acid with a higher grade of purity and at a lower price compared to traditional methods, as the reduction in byproduct substances improves the overall efficiency of the fermentation process.
Implementation Method 1
a method for producing 2-methyl-butyric acid by fermentation of a bacterium belonging to the order Enterobacterales
Data Source
AI summary
The present invention provides a method for producing 2-methyl-butyric acid by fermentation using a bacterium belonging to the order Enterobacterales which has been modified to attenuate expression of a tyrB gene encoding a protein having tyrosine aminotransferase activity. The method also allows for production of a byproduct substance of 2-methyl-butyric acid during fermentation of the Enterobacterales bacterium having 2-methyl-butyric acid-producing ability.