Mannitol is produced in a highly efficient fermentative method using Lactobacillus intermedius NRRL B-30560, or in a biochemical method using mannitol dehydrogenase isolated from this strain. Fructose serves as the primary carbon substrate in both the fermentative and biochemical conversions, but important secondary carbon sources include glucose, maltose, mannose and galactose. Mannitol is useful in the food, pharmaceutical, and medicine industries as a sweet-tasting bodying and texturing agent.