Biotin-Protein Ligase Variant 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. have limitations in effectively modifying the activity of enzymes and proteins involved in the biosynthetic pathways, necessitating further research to improve productivity.
Innovation Solution
A biotin-protein ligase variant with an amino acid substitution at position 140, specifically replacing valine with methionine, is introduced into Corynebacterium sp. to enhance the enzymatic activity, leading to increased L-glutamic acid production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional methods are used to enhance L-glutamic acid production in Corynebacterium sp., then some improvement in productivity may be achieved, but the effectiveness is limited and further research is needed to significantly improve productivity
Solution Approach 1:
The patent applies parameter changes by substituting a specific amino acid (valine to methionine at position 140) in the biotin-protein ligase to alter its enzymatic activity. This molecular-level parameter change results in significantly enhanced L-glutamic acid productivity, overcoming the limitations of conventional modification methods.
2Productivity
If genetic recombination technology is applied to modify enzymes, transcription factors and transport proteins, then L-glutamic acid productivity can be increased, but dozens of types of proteins are involved and much research is still needed
Solution Approach 1:
The patent extracts and focuses on a single critical protein (biotin-protein ligase) from the complex system of dozens of proteins involved in L-glutamic acid biosynthesis. By identifying and modifying this one key enzyme, the patent achieves significant productivity improvement without needing to systematically modify all involved proteins, thus reducing the complexity of the modification process.
Solution Approach 2:
Instead of following the conventional approach of modifying multiple downstream enzymes in the biosynthetic pathway, the patent takes an inverted approach by modifying an upstream biotin-protein ligase that is involved in carboxylation reactions. This upstream modification has cascading effects on the entire pathway, achieving efficient L-glutamic acid production with fewer modifications.
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 biotin-protein ligase variant enhances L-glutamic acid productivity by up to 12.48 times compared to the parent strain, demonstrating improved metabolic pathways and carbon source flux.
Implementation Method 1
The biotin-protein ligase may be a gene encoding biotin-protein ligase or a sequence having substantial identity thereto... functions as a carboxylase to catalyze the transfer of biotin
Implementation Method 2
L-glutamic acid is a typical amino acid that is produced by microbial fermentation... glucose mainly undergoes the glycolytic pathway (EMP), but a portion thereof is metabolized into two pyruvic acid molecules through the pentose phosphate pathway
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 biotin-protein ligase 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 biotin-protein ligase variant according to the present invention is obtained by substituting one or more amino acids in the amino acid sequence constituting biotin-protein ligase to change the enzymatic activity of the biotin-protein ligase, and a recombinant microorganism comprising the biotin-protein ligase variant is capable of efficiently producing L-glutamic acid.