Tat Protein Secretion Pathway for Corynebacterium Heterologous Production
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Solution Overview
Problem
Coryneform bacteria face difficulties in secreting certain industrially useful proteins, such as isomaltodextranase and protein transglutaminase, using the conventional Sec protein secretion pathway, which limits their industrial application.
Innovation Solution
The Tat protein secretion pathway, distinct from the Sec system, is utilized by introducing a genetic construct with a Tat system-dependent signal peptide into coryneform bacteria, enabling efficient secretion of these proteins by leveraging the Tat system's mechanism for folded protein transport.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If the Sec protein secretion pathway is used in coryneform bacteria, then the bacteria can secrete proteins, but certain industrially useful proteins such as isomaltodextranase and protein transglutaminase cannot be secreted efficiently
Solution Approach 1:
The patent changes the secretion pathway parameter from Sec to Tat system. By introducing a Tat system-dependent signal peptide sequence instead of using conventional signal peptides, the bacteria's protein secretion capability is expanded to include proteins that cannot be secreted through the Sec pathway, thereby resolving the contradiction between adaptability and productivity
Solution Approach 2:
The patent introduces a Tat system-dependent signal peptide sequence as an intermediary element that mediates the recognition and secretion of specific proteins by the Tat secretion machinery. This intermediary enables the bacteria to secrete previously non-secretable proteins like isomaltodextranase and protein transglutaminase
2Ease of manufacture
If coryneform bacteria are used for heterologous protein production, then cost and culturing efficiency are improved, but secretion capability is limited compared to Bacillus species
Solution Approach 1:
The patent makes the coryneform bacteria's secretion system more universal by enabling it to handle both Sec-pathway proteins and Tat-pathway proteins. The introduction of Tat system-dependent signal peptides allows the bacteria to perform multiple secretion functions, combining the cost advantages of coryneform bacteria with enhanced secretion capability
Solution Approach 2:
The patent changes the secretion mechanism parameter by introducing the Tat system, allowing coryneform bacteria to overcome their inherent secretion limitations while maintaining their economic advantages in terms of cost and culturing efficiency
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
This approach allows for the efficient extracellular secretion of previously difficult-to-secrete proteins like isomaltodextranase and protein-glutaminase, significantly increasing their production levels and making them more industrially viable.
Implementation Method 1
A protein secretion pathway which is completely different from the Sec system was recently discovered in the thylakoid membrane of plant cell chloroplasts... An arginine-arginine sequence is common to the signal sequences of proteins secreted through this pathway... and as a result, this pathway has come to be referred to as the Tat system (Twin-Arginine Translocation system)
Data Source
AI summary
The present invention provides a method for efficiently producing an industrially useful protein in coryneform bacteria, and more particularly, a method for efficiently producing a protein for which secretion was difficult using conventional protein secretion pathways. In particular, the present invention provides a method for efficiently producing heterologous proteins comprising culturing coryneform bacteria containing an genetic construction containing a promoter sequence which functions in coryneform bacteria, a nucleic acid sequence encoding a Tat system-dependent signal peptide region, and a nucleic acid sequence encoding a heterologous protein, in the direction from 5′-end to 3′-end, and secretory producing the heterologous protein by coryneform bacteria.