Dextransucrase is one of the important extracellular enzyme produced by majority of lactic acid bacterial species. The enzyme produces variety of polysaccharides which are having diverse biological functions with potential application in food, pharma and bio medical sectors. The enzyme in presence of sucrose and acceptor sugar produces prebiotic oligosaccharides which has wide application in food and baking industry. The present chapter embodies about the different bacterial species and their sources involved in the production of dextransucrase. A brief outlook of the reaction mechanism involved in the production of exopolysaccharides and oligosaccharides upon reaction with different sugars has been elaborated. Production of dextransucrase using various statistical optimization techniques is mentioned. An overview of various purification and downstream processing aspects with special emphasis on strain development and enhancing the reusability of enzyme by immobilization techniques has been discussed. Further, the application of enzyme in food, dairy and pharmaceutical industries has been described.