Biocatalysis has become a crucial tool in chemical synthesis, enabling the production of complex molecules with high activity, selectivity, and low environmental impact. The field has advanced significantly due to key discoveries in protein engineering, bioinformatics, recombinant technology, and DNA sequencing. This review explores enzyme engineering strategies and high-throughput screening methods that have enabled the discovery and development of enzymes for synthetic applications. Landmark developments in the field are highlighted, showcasing the diverse synthetic applications of enzymes in pharmaceutical, agricultural, food, and chemical industries. The design and development of artificial biocatalytic cascades are also examined. Enzymes are biological catalysts that accelerate chemical reactions in biological systems. Their active site allows precise positioning of substrates, enabling high selectivity and efficiency. The discovery of enzymes began in the 19th century, with key milestones including the identification of diastase by Payen and Persoz, and the 1946 Nobel Prize for proving enzymes are proteins. Mechanistic studies and advances in DNA technology have greatly enhanced our understanding of enzymes, allowing their engineering for desired applications. Enzyme engineering methods, such as rational design, directed evolution, and semi-rational design, have been developed to improve enzyme properties. Directed evolution, in particular, has proven effective in generating highly efficient catalysts by introducing random mutations and screening for improved variants. High-throughput screening (HTS) and ultrahigh-throughput screening (uHTS) methods are essential for identifying optimal enzyme variants, with techniques such as spectrophotometric, colorimetric, and fluorescence screening being widely used. In synthetic chemistry, biocatalysts are increasingly used alongside other asymmetric catalysts. Enzymes enable asymmetric synthesis, kinetic resolution, dynamic kinetic resolution, and deracemisation, which are vital for producing chiral compounds. P450 monooxygenases, transaminases, and imine reductases are key enzymes in these processes, with directed evolution and rational design playing critical roles in their optimization. Biocatalysis offers environmentally friendly and efficient methods for chemical synthesis, making it a promising area for future research and application.Biocatalysis has become a crucial tool in chemical synthesis, enabling the production of complex molecules with high activity, selectivity, and low environmental impact. The field has advanced significantly due to key discoveries in protein engineering, bioinformatics, recombinant technology, and DNA sequencing. This review explores enzyme engineering strategies and high-throughput screening methods that have enabled the discovery and development of enzymes for synthetic applications. Landmark developments in the field are highlighted, showcasing the diverse synthetic applications of enzymes in pharmaceutical, agricultural, food, and chemical industries. The design and development of artificial biocatalytic cascades are also examined. Enzymes are biological catalysts that accelerate chemical reactions in biological systems. Their active site allows precise positioning of substrates, enabling high selectivity and efficiency. The discovery of enzymes began in the 19th century, with key milestones including the identification of diastase by Payen and Persoz, and the 1946 Nobel Prize for proving enzymes are proteins. Mechanistic studies and advances in DNA technology have greatly enhanced our understanding of enzymes, allowing their engineering for desired applications. Enzyme engineering methods, such as rational design, directed evolution, and semi-rational design, have been developed to improve enzyme properties. Directed evolution, in particular, has proven effective in generating highly efficient catalysts by introducing random mutations and screening for improved variants. High-throughput screening (HTS) and ultrahigh-throughput screening (uHTS) methods are essential for identifying optimal enzyme variants, with techniques such as spectrophotometric, colorimetric, and fluorescence screening being widely used. In synthetic chemistry, biocatalysts are increasingly used alongside other asymmetric catalysts. Enzymes enable asymmetric synthesis, kinetic resolution, dynamic kinetic resolution, and deracemisation, which are vital for producing chiral compounds. P450 monooxygenases, transaminases, and imine reductases are key enzymes in these processes, with directed evolution and rational design playing critical roles in their optimization. Biocatalysis offers environmentally friendly and efficient methods for chemical synthesis, making it a promising area for future research and application.