Enzymes are biological catalysts that accelerate chemical reactions in living organisms. They are essential for numerous processes in cells, including metabolism, DNA replication, protein synthesis, and signal transduction. Enzyme biotechnology harnesses the power of enzymes for practical applications across diverse industries. By understanding and manipulating enzyme properties, researchers design biocatalysts for processes ranging from food production to environmental remediation. Enzymes offer advantages such as specificity, efficiency, and environmental friendliness, making them invaluable tools in biotechnology. Applications include enzyme-based detergents for efficient stain removal, biofuels production through enzymatic hydrolysis of biomass, and pharmaceutical synthesis with enzyme-mediated reactions. As the demand for sustainable and eco-friendly solutions grows, enzyme biotechnology continues to innovate, offering novel approaches to address complex challenges in agriculture, healthcare, and industrial manufacturing. This book is a compilation of chapters that discuss the most vital concepts and applications in the field of enzyme biotechnology. The topics covered herein deal with the core subject of enzyme biotechnology. This book is an essential guide for both academicians and those who wish to pursue this discipline further.