Chemoenzymatic Synthesis of Trehalose Analogues: Rapid Access to Chemical Probes for Investigating Mycobacteria

99 % by HPLC) in a single step from readily available glucose analogues. To demonstrate the utility of this method in mycobacteria research, we performed a simple “one‐pot metabolic labeling” experiment that accomplished probe synthesis, metabolic labeling, and imaging of M. smegmatis in a single day with only TreT and commercially available materials. } data-sheets-userformat= { 2 :771, 3 :{ 1 :0}, 4 :[null,2,16777215], 11 :4, 12 :0} style= font-size: 10pt; font-family: Arial; \u3eTrehalose analogues are emerging as valuable tools for investigating Mycobacterium tuberculosis , but progress in this area is slow due to the difficulty in synthesizing these compounds. Here, we report a chemoenzymatic synthesis of trehalose analogues that employs the heat‐stable enzyme trehalose synthase (TreT) from the hyperthermophile Thermoproteus tenax . By using TreT, various trehalose analogues were prepared quickly (1 h) in high yield (up to \u3e99 % by HPLC) in a single step from readily available glucose analogues. To demonstrate the utility of this method in mycobacteria research, we performed a simple “one‐pot metabolic labeling” experiment that accomplished probe synthesis, metabolic labeling, and imaging of M. smegmatis in a single day with only TreT and commercially available materials