Microorganisms from polar environments can play crucial roles in the biotechnological production of enzymes of pharmacological interest. In this chapter, we compile the main studies focused on the biotechnological aspects of enzymes derived from microorganisms from Antarctica and the Arctic with human pharmacological applications. Enzymes from microorganisms in these environments may exhibit catalytic activity at low and moderate temperatures. With the advancement of modern biotechnology, cold-adapted enzymes have been widely used in food production, animal feed, the textile industry, and especially in the development of new drugs. Consequently, these enzymes have garnered much attention in recent years, driven by the advent of bioinformatics and heterologous expression in genetically modified organisms (GMO). Enzymes such as L-asparaginase, β- and α-galactosidases, lipase, and superoxide dismutase stand out in the production and potential formulation of new biopharmaceuticals. Genetically manipulated strains stimulated to produce specific pharmacologically relevant enzymes, combined with bioinformatics, may become crucial in a variety of biotechnological applications. Thus, enzymes produced by polar microorganisms with pharmaceutical interest may lead to a larger annual market than thermostable enzymes in the near future.