Bioactive peptides (BPs) play key roles in regulating cellular metabolism, and are therefore of special interest to the nutraceutical and pharmaceutical industries. Protein digestion is a major route to production of BPs. Biocatalyst-aided processes are advantageous in that they generate protein hydrolysates with high specificity that are non-immunogenic and resistant to proteolysis in the gastrointestinal tract. On the other hand, these approaches are limited by their high production cost and poor diversity of peptides, as well as the requirement for sophisticated analysis for functional validation and scale-up. The recent development of microbial genome and protein databases, together with in silico analysis, have enabled BP design with bioavailability and the identification of new routes for the isolation of BPs from natural resources. In this review, we focus on recent developments in native BP targeting informatics, novel microbial protease mining, and screening methods, which together will accelerate the development of genome-based microbial routes to therapeutic BPs that promote human health and nutrition.