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Corner Engineering: Tailoring Enzymes for Enhanced Resistance and Thermostability in Deep Eutectic Solvents
Xinyue Wang1, Yijie Sheng1, Haiyang Cui2,3
1School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, No. 2 Xuelin Road, Nanjing, 210097, China.
Angewandte Chemie (International Ed. in English)
|November 27, 2023
Summary
Protein engineering via corner engineering enhances enzyme resistance to deep eutectic solvents (DESs) and improves thermostability. This strategy yields more robust enzymes for biocatalysis and biotransformation applications.
Area of Science:
- Biocatalysis and enzyme engineering
- Green chemistry and sustainable solvents
Background:
- Deep eutectic solvents (DESs) offer advantages in synthesis and safety but often reduce enzyme activity and stability.
- Developing robust enzymes is crucial for expanding the application of DESs in biocatalysis.
Purpose of the Study:
- To develop a general protein engineering strategy, termed corner engineering, to create enzymes resistant to DESs and enhance their thermostability.
- To validate the efficacy of corner engineering on industrial enzymes for biocatalysis and biotransformation.
Main Methods:
- Applied corner engineering to Bacillus subtilis lipase A (BSLA) by tailoring the transition region of its structure.
- Created and characterized multiple DES-resistant BSLA variants with improved thermostability.
- Validated the approach on endo-β-1,4-glucanase PvCel5A and esterase Bs2Est.
Main Results:
- Engineered BSLA variants showed significant increases in catalytic efficiency (up to 10.0-fold) in DESs and enhanced thermal resistance (up to 6.7-fold).
- The strategy was successfully applied to PvCel5A and Bs2Est, demonstrating its generality.
- Molecular investigations identified increased water molecules at the substrate binding cleft and finetuned helix formation as key factors.
Conclusions:
- Corner engineering is an effective strategy for developing DES-resistant and thermostable enzymes.
- This approach provides molecular insights into enzyme-DES interactions, guiding rational enzyme design for industrial applications.
