The chapter relates to new knowledge created in a specialized area of biotechnology, biocatalysis, where enzymes are used as environmentally benign catalysts for reactions producing valued chemical products or processes. It integrates enzyme discovery with various related fields, including chemistry, biochemistry, microbiology, molecular biology, genomics, and engineering. I describe research on various reaction systems and the relevance of new discoveries of novel materials and bioprocesses. New technologies have enabled discovery to take place in an ongoing cycle. Identifying the biological agent, the enzyme, creates new knowledge in biotech processes. Here is where techniques and technologies have evolved through some giant leaps. From an era of sequencing DNA isolated from cultured cells, biocatalyst discovery has moved to current times when next-generation sequencing allows collection and analysis of huge bodies of complex biological data, CRISPR gene editing enables the design and production of novel biological agents, and digital technologies incorporating AI enable creative new ideas; the discovery of new enzymes to use as biocatalysts has even greater scope for progress.