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Strategy and implementation of a system for protein engineering
D W Goddette1, T Christianson, B F Ladin
1COGNIS Inc., Santa Rosa, California 95407.
Journal of Biotechnology
|March 1, 1993
Summary
This study enhanced protein stability using rational design and protein engineering methods. The approach successfully increased subtilisin BL enzyme stability over 10-fold at 50°C.
Area of Science:
- Protein Engineering
- Enzyme Stability
- Biotechnology
Background:
- Rational design is crucial for protein engineering when structural and literature data are available.
- Developing rapid and accurate methods for mutagenesis, expression, verification, purification, and characterization is essential for enzyme modification.
Purpose of the Study:
- To outline an industrial protein engineering project from inception to completion.
- To demonstrate a rational design strategy for improving enzyme stability.
Main Methods:
- Utilized a three-dimensional crystal structure and literature data to define a rational design strategy.
- Employed methods for mutagenesis, expression, verification, purification, and characterization of mutant enzymes.
- Refined the design strategy throughout the project.
Main Results:
- Successfully increased the stability of subtilisin BL by over 10-fold at 50°C.
- Achieved an overall success rate greater than 60% in the protein engineering project.
Conclusions:
- The described protein engineering approach is effective for enhancing enzyme stability.
- This methodology can be applied to improve the stability of other proteins and enzymes.