Corynebacterium IMP Export Protein Variant

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Solution Overview

Problem

Current methods for producing 5'-inosine monophosphate (IMP) through microbial fermentation face challenges in achieving high yields due to suboptimal enzyme properties and unclear structure-function correlations, limiting effective enzyme modification and enhancement techniques.

Innovation Solution

Development of a protein variant with the activity of exporting IMP, specifically from the genus Corynebacterium, which includes polynucleotides encoding these proteins and vectors for microorganisms, enhancing the export activity to increase IMP production.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If rational design and site-directed mutagenesis of enzymes are applied to improve enzyme function, then enzyme activity can be enhanced, but information on the structure of target enzymes is insufficient or the structure-function correlation is not clear, preventing effective application

Engineering Contradiction:
Improveenzyme functionVSAvoidstructure-function correlation
Core Design Contradiction:
ReliabilityVSLoss of information

Solution Approach 1:

The patent applies parameter changes by systematically modifying amino acid sequences at specific positions (e.g., positions 164, 222, 64) to optimize enzyme function. Through directed evolution and site-directed mutagenesis, the invention changes physical-chemical parameters of the enzyme structure to improve IMP export activity while overcoming insufficient structure-function correlation information.

Inventive Principle:
Principle #35Parameter changes

2Reliability

If directed evolution method is used to screen enzymes of desired traits from a library of modified enzymes, then enzyme activity can be improved, but the process requires construction of large enzyme libraries and extensive screening

Engineering Contradiction:
Improveenzyme activityVSAvoidenzyme library construction
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The patent applies local quality by focusing mutations at specific critical positions (e.g., positions 164, 222, 64) rather than random mutagenesis throughout the entire enzyme. This targeted approach to local regions with high impact on function reduces the complexity of library construction while maintaining effective screening for desired traits.

Inventive Principle:
Principle #3Local quality

3Quantity of substance

If microbial fermentation is used to produce IMP, then IMP can be produced through metabolic pathways, but the IMP export efficiency is suboptimal due to unclear structure-function correlations of export proteins

Engineering Contradiction:
ImproveIMP productionVSAvoidIMP export efficiency
Core Design Contradiction:
Quantity of substanceVSReliability

Solution Approach 1:

The patent applies feedback by measuring IMP export efficiency and using this information to guide further protein optimization. Through iterative cycles of mutation, expression, and activity assessment, the invention refines the export protein's amino acid sequence to improve IMP production yield, creating a feedback loop that continuously enhances export efficiency.

Inventive Principle:
Principle #23Feedback

Data Source

PatentUS11987611B2Polypeptide and method of producing IMP using the same
Publication Date: 2024.05.21 CJ CHEILJEDANG CORP

AI summary

The present disclosure relates to a novel polypeptide having an activity of exporting 5′-inosine monophosphate, a microorganism comprising the same, a method for preparing 5′-inosine monophosphate using the same, and a method for increasing export of 5′-inosine monophosphate.