In the realm of enzymatic research, strides have been made in the design of enzymes tailored for novel substrate classes, marking a pivotal juncture in the pursuit of biocatalytic applications. Notably, recent advancements underscore the expanding scope of enzyme design, reaching beyond traditional substrates to address the catalytic challenges posed by emerging classes. Enzyme engineering strategies have harnessed computational approaches and directed evolution to fine-tune the catalytic machinery, enabling the recognition and transformation of previously untapped substrates with precision. The exploration of diverse substrate classes, ranging from synthetic compounds to naturally occurring molecules, has propelled the field toward unprecedented versatility. Furthermore, the integration of structural bioinformatics has played a pivotal role in deciphering the intricate interactions governing substrate specificity. As researchers delve into uncharted enzymatic territories, the convergence of computational modeling, high-throughput screening, and innovative design methodologies illuminates the trajectory toward a new era of enzymatic exploration and application.