Synthesis of trehazolin β-anomer (3) from a D-glucose derived azido alcohol (4), was accomplished. 2-Chloro-l-methylpyridinium iodide was used in place of 2-chloro-3-ethylbenzoxazolium tetrafluoroborate as a means of preventing concomitant anomerization. Evaluation of compound (3) reveals that the stereochemistry of the anomeric position is significant for generation of inhibitory activities towards trehalases.