Phosphorolytic enzymes that act on carbohydrates are interesting enzymes not only in terms of their basic properties but also in regard to their various applications. However, they have not been studied in as much detail as other types of enzymes such as the hydrolases and the synthases. Phosphorolytic enzymes have attracted attention because of their application in the synthesis of carbohydrate chains, exploiting their strict regiospecificities. In addition, a number of new phosphorolytic enzymes have been discovered recently. We have been investigating the phosphorolytic enzymes and have discovered their unique characters and applications. In this paper, the following are described: 1. Analyses of the reaction characterized by celobiose phosphorylase from Cellvibrio gilvus; 2. Characterization of laminaribiose phosphorylase from Euglena gracilis; 3. A specific colorimetric quantification of cellobiose employing cellobiose phosphorylase; 4. Synthesis of laminarioligosaccharides; 5. Conversion of sucrose into cellobiose using two phosphorolytic enzymes.