University of Southern Maine 1 article published in JoVE Biochemistry Rapid One-step Enzymatic Synthesis and All-aqueous Purification of Trehalose Analogues Lisa M. Meints1, Anne W. Poston1, Brent F. Piligian1, Claire D. Olson1, Katherine S. Badger2, Peter J. Woodruff2, Benjamin M. Swarts1 1Department of Chemistry and Biochemistry, Central Michigan University, 2Department of Chemistry, University of Southern Maine Trehalose analogues are emerging as important molecules for bio(techno)logical and biomedical applications. We describe an optimized protocol for enzymatically synthesizing and purifying trehalose analogues that is simple, efficient, fast, and environmentally friendly. Its application to the rapid production and administration of a probe for the detection of mycobacteria is demonstrated.