Carotenoids are lipophilic terpenoid pigments with biotechnological applications. They are ubiquitous in photosynthetic species and are also produced by many non-photosynthetic bacteria, yeasts, and filamentous fungi, which are amenable to genetic manipulations and can be easily cultured in large fermenters. The biotechnological interest of carotenoids derives from their healthy properties due to their antioxidant activity or to physiological functions. Some carotenoids can be used as a source of vitamin A, chromophore of visual opsins and the origin of the retinoid metabolism associated with gene regulation functions in chordates, and others are related to the maintenance of the macula in the eye. In general, consumption of carotenoids is associated with a decrease in the possibilities of suffering from numerous diseases, from cancer to alterations in the immune system. Fungal carotenoid biosynthetic pathways are relatively simpler than those found in photosynthetic organisms, especially in plants, and not many carotenoids are usually found in these organisms. These include carotenes, such as lycopene and β-carotene, and xanthophylls, such as astaxanthin, neurosporaxanthin, or torularhodin. Some of these carotenoids are produced industrially, but it is to be hoped that their applied use will be extended to others. This chapter reviews the biotechnological and production possibilities of the best-known fungal carotenoids. The scientific literature on this subject is very extensive, especially in the areas of the biological effects of carotenoids on humans and the improvement of production for biotechnological purposes. In this review, we select significant contributions, recent or historical, that serve to make a comprehensive review of fungal carotenoids, focusing on their actual or potential production and their possible applications.