Glutamine Synthetase Vector to Reduce ATase Feedback Inhibition
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Solution Overview
Problem
Existing methods for increasing glutamine production are limited by feedback inhibition of glutamine synthetase (GS) due to glutamine synthetase adenylyltransferase (ATase), which reduces productivity.
Innovation Solution
A recombinant vector containing a nucleotide sequence encoding a glutamine synthetase with a specific amino acid sequence (SEQ ID NO: 1) from Corynebacterium glutamicum strain KFCC10694, combined with a promoter and transcription terminator, is used to transform a strain, reducing the impact of ATase inhibition and increasing glutamine productivity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If nitrogen source is supplied to produce glutamine, then glutamine production increases, but GS activity is feedback-inhibited by ATase, resulting in reduction in glutamine production
Solution Approach 1:
The invention extracts and removes the harmful regulatory elements (glnB and glnE genes encoding PII and ATase) from the system. By deleting these genes, the feedback inhibition mechanism is completely eliminated, allowing GS to function without regulation and continuously produce glutamine from supplied nitrogen sources.
Solution Approach 2:
The invention changes the regulatory parameters of GS by modifying the genetic background. Specifically, it alters the presence/absence state of regulatory genes (glnB, glnE) to change the system from a regulated state to an unregulated state, thereby changing GS activity parameters from controlled to constitutive high-level expression.
2Productivity
If glnB and glnE genes are inhibited to increase glutamine production, then glutamine productivity improves, but further improvement is limited
Solution Approach 1:
The invention completely removes the regulatory genes (glnB and glnE) from the genome through deletion, rather than merely inhibiting their expression. This extraction of harmful regulatory elements achieves maximum productivity improvement by eliminating feedback inhibition entirely, going beyond partial inhibition approaches.
Solution Approach 2:
Instead of trying to enhance GS activity directly or partially inhibit regulators, the invention inverts the approach by completely deleting the regulatory genes. This inversion transforms the system from one with active regulation to one without regulation, achieving superior productivity.
3Quantity of substance
If GS activity is increased to enhance glutamine production, then glutamine yield improves, but feedback inhibition by supplied nitrogen source reduces efficiency
Solution Approach 1:
The invention converts the harmful feedback inhibition mechanism into a benefit by completely removing the regulatory genes. The supplied nitrogen source, which previously caused feedback inhibition and reduced efficiency, now serves purely as substrate for continuous glutamine production without any regulatory interference.
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 transformed strain significantly enhances glutamine production by minimizing feedback inhibition, resulting in increased glutamine yield.
Implementation Method 1
The glutamine synthetase is an enzyme that synthesizes glutamine from glutamate and ammonia
Data Source
AI summary
The present invention relates to a strain which has increased glutamine productivity due to being transformed with a vector containing a nucleotide sequence that encodes a glutamine synthetase consisting of the amino acid sequence of SEQ ID NO: 1.