Enterobacter cloacae NRRL B-23289, isolated from decaying wood/corn soil samples by the USDA Agricultural Research Service (Peoria, IL), is a natural producer of 2,3-BD. Previous work at the USDA ARS has shown that this strain is more efficient in converting ketose than aldose sugars to 2,3-BD. Particularly interesting is that fermentation of fructose showed higher 2,3-BD yield within a much shorter period of time as compared to glucose. We have previously developed a method of simultaneous isomerization and reactive extraction (SIRE) to produce the ketose isomer of xylose (xylulose) in high yield and purity. Using this innovative method to produce large quantities of nearly pure xylulose, we evaluated 2,3-BD production from the NRRL B-23289 strain.