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Engineering Bacterial Laccase with Improved Catalytic Activity and Thermostability by Rational Design
Xuting Sun1,2, Xiaofan Lin1, Yufan Xian1
1College of Life Science and Technology, Beijing University of Chemical Technology, No. 15 Beisanhuan Road East, Chaoyang District, Beijing, 100029, China.
Applied Biochemistry and Biotechnology
|May 9, 2025
Summary
Engineered Bacillus subtilis laccase CotA (DTA) shows improved activity and stability. This enhanced enzyme efficiently degrades mycotoxins and aids lignocellulose pretreatment for industrial applications.
Area of Science:
- Biotechnology
- Enzyme Engineering
- Industrial Microbiology
Background:
- Laccases are crucial multi-copper oxidases with diverse industrial uses.
- Bacillus subtilis laccase CotA is a well-studied enzyme with potential for improvement.
Purpose of the Study:
- To engineer Bacillus subtilis laccase CotA for enhanced catalytic activity and thermostability.
- To investigate the structural and functional basis for the improved enzyme properties.
Main Methods:
- Rational design and iterative mutation of CotA laccase.
- Characterization of enzyme activity, thermostability, and degradation capabilities.
- Assessment of lignocellulose pretreatment efficiency.
Main Results:
- A triple mutant (DTA) exhibited a 2.7-fold increase in catalytic activity and a 1.4-fold increase in thermostability compared to wild-type CotA.
- Enhanced DTA activity is linked to strengthened active site interactions; improved stability is due to increased hydrophobic residues and surface flexibility.
- DTA demonstrated superior degradation of aflatoxin B1 and ZEN mycotoxins and more effective lignocellulose pretreatment.
Conclusions:
- The engineered DTA laccase presents significantly improved catalytic performance and stability.
- DTA shows promising potential for industrial applications, including mycotoxin bioremediation and biomass processing.
