Stabilized Transglutaminase Pro-Sequence Peptide Binding
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Solution Overview
Problem
Despite existing stabilization techniques, transglutaminases still require additional components to maintain stability, particularly in food applications, where minimal extra additions are desirable.
Innovation Solution
A stabilized transglutaminase is produced by binding a pro-sequence peptide to mature transglutaminase, enhancing stability against pH, temperature, oxidation, and storage conditions, using a microorganism like Streptomyces mobaraensis and specific amino acid sequences.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If additional stabilizers (organic acid, inorganic acid, polyphenol, thiol, sugar alcohol, or protein) are added to transglutaminase, then stability during storage is improved, but the complexity of the system and the number of extra components increase
Solution Approach 1:
The transglutaminase molecule itself provides stabilization through its pro-form structure. The pro-sequence peptide remains bound to the mature transglutaminase, creating a self-stabilizing system that does not require external stabilizers. This self-service approach resolves the contradiction by making the enzyme self-sufficient for stability while minimizing extra components.
Solution Approach 2:
The pro-form transglutaminase represents a composite structure where the pro-sequence peptide and mature transglutaminase function together as an integrated stabilizing system. This internal composite structure provides stability without requiring separate stabilizing agents, thus resolving the contradiction between stability improvement and component complexity.
2Reliability
If transglutaminase is used in food applications, then protein binding and food manufacturing are improved, but the risk of unexpected circumstances increases without sufficient stabilization
Solution Approach 1:
The pro-sequence peptide acts as a protective cushion that remains bound to the mature transglutaminase before food processing applications. This pre-existing protective structure cushions the enzyme against destabilizing conditions during storage and application, ensuring reliable performance while minimizing risks of unexpected circumstances.
Solution Approach 2:
The invention changes the structural parameter of transglutaminase by maintaining the pro-form with bound pro-sequence peptide rather than using the mature form alone. This parameter change (structural configuration) fundamentally alters the stability profile, providing enhanced reliability in food applications without requiring additional stabilizing parameters or components.
3Reliability
If mature transglutaminase is used, then enzymatic activity is achieved, but stability against oxidation, temperature, and pH is poor
Solution Approach 1:
The invention merges the pro-sequence peptide with the mature transglutaminase in a stable complex structure. This merging creates a unified molecule where the pro-sequence peptide and mature enzyme work together, combining the enzymatic activity of the mature form with the stabilizing properties of the pro-sequence, thus resolving the contradiction between activity and stability.
Solution Approach 2:
The pro-sequence peptide acts as an intermediary element that mediates between the mature transglutaminase and the destabilizing environmental factors (oxidation, temperature, pH). This intermediary structure protects the active site and overall enzyme structure from harmful effects while maintaining enzymatic function, resolving the contradiction between activity and stability.
Data Source
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AI summary
Disclosed is a transglutaminase having excellent stability. Also disclosed is a process for producing the transglutaminase. Specifically disclosed is a stabilized transglutaminase, which has such a structure in which a pro-sequence peptide of transglutaminase is bound to a mature transglutaminase. Also specifically disclosed is a process for producing stabilized transglutaminase, which includes the steps of: culturing a microorganism capable of producing transglutaminase under the conditions where transglutaminase can be produced; and separating and collecting matured transglutaminase having a pro-sequence peptide bound thereto from a culture medium.