Agar, a major component of the cell wall of red algae, is an interesting heteropolysaccharide containing an unusual sugar, 3,6-anhydro-l-galactose. It is widely used as a valuable material in various industrial and experimental applications due to its characteristic gelling and stabilizing properties. Agar-derived oligosaccharides or mono-sugars produced by various agarases have become a promising subject for research owing to their unique biological activities, including anti-obesity, anti-diabetic, immunomodulatory, anti-tumor, antioxidant, skin-whitening, skin-moisturizing, anti-fatigue, and anti-cariogenic activities. Agar is also considered as an alternative sustainable source of biomass for chemical feedstock and biofuel production to substitute for the fossil resource. In this review, we summarize various biochemically characterized agarases, which are useful for industrial applications, such as neoagarooligosaccharide or agarooligosaccharide production and saccharification of agar. Additionally, we succinctly discuss various recent studies that have been conducted to investigate the versatile biological activities of agar-derived saccharides and biofuel production from agar biomass. This review provides a basic framework for understanding the importance of agarases and agar-derived saccharides with broad applications in pharmaceutical, cosmetic, food, and bioenergy industries.