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High temperature increases the refolding yield of reduced lysozyme: implication for the productive process for
Ryusuke Sakamoto1, Shingo Nishikori, Kentaro Shiraki
1School of Materials Science, Japan Advanced Institute of Science and Technology (JAIST), 1-1 Asahidai, Tatsunokuchi, Ishikawa 923-1292, Japan.
Biotechnology Progress
|August 7, 2004
Summary
High temperatures significantly improve protein refolding yields from inclusion bodies using a simple dilution method. This thermal-assisted refolding offers a cost-effective solution for producing active proteins in biotechnology.
Area of Science:
- Biotechnology
- Protein Chemistry
- Biophysics
Background:
- Protein misfolding and aggregation into inclusion bodies is a major challenge in recombinant protein production.
- Obtaining active proteins from inclusion bodies often requires complex and inefficient refolding processes.
Purpose of the Study:
- To investigate the effect of high temperature on the refolding yield of reduced lysozyme from inclusion bodies.
- To explore the potential of thermal-assisted refolding as a simple and cost-effective method for protein refolding.
Main Methods:
- Inclusion bodies of reduced lysozyme were prepared.
- Refolding was induced by a simple dilution method at different temperatures (98°C and 20°C).
- Refolding yields were quantified and compared between the different temperature conditions.
Main Results:
- High temperature (98°C) significantly increased the refolding yield of reduced lysozyme by approximately three times compared to refolding at room temperature (20°C).
- The thermally unfolded state of lysozyme was found to be a more productive intermediate for folding than the denaturant-induced fully unfolded state.
- The simple dilution method combined with high temperature proved effective for refolding.
Conclusions:
- Thermal-assisted refolding is a highly effective strategy for increasing protein refolding yields from inclusion bodies.
- This method is simple, cost-effective, and applicable to various reduced and denatured proteins.
- High-temperature unfolding provides a more favorable state for subsequent protein refolding.