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Isothermal Compressibility Perturbation as a Protein Design Principle for T1 Lipase Stability-Activity Trade-Off
Nan Zheng1, Ling Gao1, Mengfei Long1
1Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, China.
Journal of Agricultural and Food Chemistry
|April 21, 2023
Summary
Enzyme engineering achieved high stability and activity using isothermal compressibility perturbation engineering (ICPE). This method improved T1 lipase performance, demonstrating a new strategy for enzyme evolution.
Area of Science:
- Biochemistry
- Enzyme Engineering
- Protein Science
Background:
- Enzyme evolution faces a stability-activity trade-off, limiting the development of highly effective enzymes.
- Understanding the mechanism behind this trade-off is crucial for advancing enzyme engineering.
Purpose of the Study:
- To investigate the stability-activity seesaw-like mechanism using isothermal compressibility perturbation engineering (ICPE).
- To engineer a prototypical enzyme, T1 lipase, for enhanced stability and catalytic activity.
Main Methods:
- Employed isothermal compressibility perturbation engineering (ICPE) to generate enzyme mutants.
- Analyzed the correlation between stability and activity in the engineered mutants.
- Characterized the best performing variant for thermal stability, catalytic activity, and solvent resistance.
Main Results:
- A high Pearson correlation (r = 0.93) was observed between stability and activity in ICPE-derived mutants.
- The best variant (A186L/L188M/A190Y) showed a Tm of 78.70 °C, activity of 474.04 U/mg, and 73.33% increased DMSO resistance.
- An elastic activation mechanism involving conformational changes was proposed to explain the stability-activity balance.
Conclusions:
- The ICPE strategy provides a comprehensive understanding of the enzyme stability-activity trade-off.
- This approach significantly enhances enzyme performance, offering a valuable tool for enzyme engineering applications.
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