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Updated: Sep 12, 2025

A New Screening Method for the Directed Evolution of Thermostable Bacteriolytic Enzymes
Published on: November 7, 2012
Computational-assisted protein engineering to develop thermostable and highly active catalase for industrial and
Shuai Xu1, Ya-Min Chen1, Xiang-Yu Meng1
1State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 200237 Shanghai, China.
Abstract:
Catalases are ideal biocatalysts for hydrogen peroxide removal in industrial wastewater and enzymatic processes, yet the practical application is hindered by poor thermostability. In this study, a highly active catalase was identified from Fictibacillus enclensis using a computer-assisted screening strategy. To overcome thermal instability, a computational design framework integrating ProteinMPNN-based sequence optimization with physics-based energy calculations was developed. The engineered variant, D78P/K201R/E384Y/T435A, exhibited a 1.9-fold increase in catalytic efficiency and a 4.9-fold extension of half-life at 40°C. Molecular dynamics simulations and structural analyses revealed that the mutations conferred enhanced global rigidity through stabilized hydrogen-bond networks. Moreover, the variant was employed for the treatment of industrial effluents containing hydrogen peroxide residues andthe biocatalytic upgrading of glycerol. This study not only paved the way for industrial applications of catalase but also established a strategic framework for enhancing both stability and catalytic activity in enzyme engineering.
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