Spl1 Promoter Enhances Corynebacterium Gene Expression
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Solution Overview
Problem
Gene expression systems in microorganisms of the genus Corynebacterium exhibit lower expression efficiency compared to those in E. coli, necessitating the development of a promoter that can induce strong gene expression.
Innovation Solution
A novel nucleic acid molecule with promoter activity, specifically the spl1 promoter, is developed and used to create a recombinant vector and microorganism for enhanced target protein production, including the production of psicose by reacting the microorganism with fructose.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional promoters (Psod, Peftu, PgapA, Pcj4) are used in Corynebacterium, then the gene expression system can function, but the expression efficiency is low compared to E. coli systems
Solution Approach 1:
The patent modifies promoter sequences by changing specific nucleotide positions (e.g., positions -10 to -15 relative to transcription start site) to optimize promoter strength. The spl1 promoter was created by modifying the Pcj4 promoter sequence, specifically changing the -10 region from TATAAT to TATGTT, which significantly enhanced transcription initiation efficiency and gene expression levels in Corynebacterium
Solution Approach 2:
The patent combines elements from different promoter sequences to create a composite promoter with superior properties. The spl1 promoter integrates optimized -10 and -35 regions from various Corynebacterium promoters along with specific spacer sequences, creating a hybrid promoter structure that achieves expression efficiency comparable to E. coli systems
2Productivity
If promoter strength is increased to enhance gene expression, then productivity improves, but control over expression levels becomes more difficult
Solution Approach 1:
The patent creates promoters with adjustable strength by designing modular structures where specific regions can be modified to tune expression levels. The spl1 promoter and its variants allow for dynamic control of gene expression by altering key nucleotide positions, enabling researchers to optimize expression levels for different application requirements while maintaining high productivity
Data Source
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AI summary
The present disclosure relates to a novel promoter, a vector comprising the promoter, a microorganism comprising the vector, and a method for producing a target protein using the promoter. The promoter obtained by the present disclosure exhibits an increased gene expression rate compared to promoters used in a large-scale production of proteins from the existing recombinant microorganisms, and can thus be used advantageously in food, pharmaceutical, and agricultural industries in which high-yield production of functional materials from microorganisms is needed.