Redesigning Robust Biocatalysts by Engineering Enzyme Microenvironment and Enzyme Immobilization

Enzymes have replaced traditional industrial catalysts as more efficient, eco-friendly, and sustainable alternatives that can be used in different biotechnological processes, food, and pharmaceutical industries. Yet, the enzymes from nature are engineered to make them adapt and enhance their durability in the industrial environment. Techniques have been developed to tailor such enzymes and overcome the hurdles of efficient bio-catalysis. Protein engineering has transformed the art of enzyme tailoring by providing opportunities to create enzymes with better functionality, such as increased stability, reaction product inhibition, and improved catalytic activity. Protein engineering and immobilization are compatible approaches used side by side to improve enzyme properties. The surge in enzyme immobilization has enabled robustness and outstanding functionality in harsh industrial conditions with high temperatures and organic solvents. The introduction of multi-enzyme catalytic cascades according to a mix of biocatalysts opens up many new possibilities in biosynthesis, biocatalysis, and biotransformation. Multi-enzyme cascade reactions often provide reaction time and cost-related advantages. Immobilization techniques and multi enzymes cascades are used to obtain robust industrial catalysts. This review focuses on the state-of-the-art strategies and trends to construct novel biocatalysts having amplified catalytic activity and substrate specificity required for industrial application. Further development in protein engineering, immobilization techniques, and multi-enzyme cascade reactions might pave the way for future industrial biocatalysis.Graphical Abstract